Journal articles: 'Monoclonal antibody probes Diagnostic use'

1

McQuaid, S., and G. M. Allan. "Detection protocols for biotinylated probes: optimization using multistep techniques." Journal of Histochemistry & Cytochemistry 40, no. 4 (April 1992): 569–74. http://dx.doi.org/10.1177/40.4.1552190.

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Recent studies using biotinylated in situ hybridization (ISH) have utilized a wide range of detection protocols for the biotinylated hybrids, leading to conflicting reports in the literature regarding sensitivity. In this study we compared 11 different detection protocols for biotinylated ISH using a measles virus-specific RNA probe on formalin-fixed, paraffin-embedded central nervous system tissue infected with measles virus. Maximum sensitivity was achieved with five-step detection protocols incorporating the use of a monoclonal antibody to biotin. Single-step detection protocols were found to be insensitive, as shown by their failure to detect viral nucleic acid in infected white-matter cells. Only by increasing the number of steps in the detection protocols were these infected cells demonstrable. Unless pre-hybridization, hybridization, and detection protocols are optimized, the results obtained in pathogenicity studies using ISH could be misinterpreted, leading to false conclusions about nucleic acid distribution. This also applies to the ever-increasing use of ISH for diagnostic purposes.

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Rosenberg, J. S., M. J. Melnicoff, and P. Wilding. "Fc receptors for IgG on human neutrophils: analysis of structure and function by using monoclonal antibody probes." Clinical Chemistry 31, no. 9 (September 1, 1985): 1444–48. http://dx.doi.org/10.1093/clinchem/31.9.1444.

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Abstract Structural and functional characteristics of Fc receptors for IgG (Fc gamma) on human neutrophils were examined with two monoclonal antibody probes specific for the Fc gamma receptors, Leu 11b and 3G8. To determine the distribution, density, and membrane mobility of the Fc gamma receptor, we used immunogold staining techniques, flow cytometry analysis, and fluorescence microscopy. Both 3G8 and Leu 11b inhibited several cell functions, thereby depicting the regulatory role of the Fc gamma receptor in mediating neutrophil activities. Among the functions studied were release of lysosomal enzymes, release of superoxide anion (O2-), and Fc-dependent rosette formation and phagocytosis. The densities of Fc gamma determinants recognized by Leu 11b and 3G8 on cells from a patient with chronic myelogenous leukemia were less than the density of epitopes on neutrophils from a normal individual. Taken together, the detailed analysis of physical and functional aspects of the Fc gamma receptor on neutrophils described in this study serve as a model for further assessment of the use of Fc gamma phenotyping of cells as a diagnostic tool.

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Parker, Edward, Christina MacLaughlin, Samantha Wala, Gilbert Walker, and Chen Wang. "Targeting CLL Cells Using Rituximab-Conjugated Surface Enhanced Raman Scattering (SERS) Gold Nanoparticles." Blood 116, no. 21 (November 19, 2010): 2691. http://dx.doi.org/10.1182/blood.v116.21.2691.2691.

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Abstract Abstract 2691 Measuring the expression of cell surface markers is crucial to the effective diagnosis of lymphoproliferative disorders such as chronic lymphocytic leukemia (CLL). However, conventional fluorescent probes are constrained by the degradative effect of photobleaching and the broad emission spectra of dyes, which restricts the multiplexing capacity of marker detection. Recent developments in nanoparticle-based technology may confer significant advantages over these traditional tools. In particular, surface enhanced Raman scattering (SERS) nanoparticles (NPs) can now be targeted to cells via conjugation to monoclonal antibodies. These particles are composed of colloidal gold cores surrounded by an organic dye with a distinct Raman-scattering signature. In addition to providing stable long-term signals, Raman probes typically exhibit spectral bands more than 30 times narrower than those of fluorescence techniques, thereby greatly increasing the multiplexing potential of phenotypic analysis. In this study, we developed SERS NPs conjugated to the monoclonal antibody rituximab in order to target the surface marker CD20. Rituximab has been established as an effective therapeutic antibody in the treatment of several B-cell disorders, though its precise mechanism of action is unclear. The preparation of SERS probes was achieved by coating 60 nm gold particles with the Raman-active reporter malachite green isothiocyanate (MGITC) followed by a stabilizing layer of polyethylene glycol (PEG). These particles were then covalently linked to rituximab using ethyl dimethylaminopryl carbiimide (EDC) and sulfo-NHS chemistry. Following the incubation of CLL cells with rituximab conjugates, samples were examined using darkfield microscopy, and Raman scatter analyzed using a Raman spectroscope. The resulting spectra were concordant with the successful retention of SERS probes, as indicated by an increase in the intensity of MGITC Raman peaks as the staining concentration of conjugates increased. However, the significant background signal obtained using unconjugated control NPs highlights the necessity to incorporate more rigorous steps to remove unbound particles in future studies. Darkfield imaging strongly confirmed the successful binding of SERS probes to CLL cells, which notably failed to retain control NPs. Conjugate targeting was also disrupted by blocking CD20 binding sites with unconjugated rituximab prior to SERS probe staining, thereby confirming that NP targeting was not the product of non-specific binding. Together, these results strongly indicate the successful incorporation of a therapeutic antibody into the NP-based targeting of CD20. In conjunction with SERS probes directed at other markers, this novel diagnostic approach could have a profound impact on the multiplexing capacity of cell surface marker detection during the diagnosis of lymphoproliferative disorders. In addition, the long-term stability of these probes might facilitate the use of NP conjugates as tracers to examine the effects of rituximab binding, thereby providing valuable insight into the mechanisms of antibody-based immunotherapy. Disclosures: No relevant conflicts of interest to declare.

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Ruppel, Andreas, Ute Breternitz, and Reinhard Burger. "Diagnostic Mr 31 000 Schistosoma mansoni proteins: requirement of infection, but not immunization, and use of the “miniblot” technique for the production of monoclonal antibodies." Journal of Helminthology 61, no. 2 (June 1987): 95–101. http://dx.doi.org/10.1017/s0022149x00009810.

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ABSTRACTAntibodies directed against diagnostic Mr 31 000 polypeptide(s) of adult Schistosoma mansoni were already formed in mice during prepatency. In contrast, repeated immunization of mice with homogenates of adult schistosomes failed to elicit antibodies detectable in immunoblots in the Mr 31 000 region. Therefore, spleen cells of infected mice were used to produce hybridoma lines. The “miniblot technique” was developed in order to detect in hybridoma supernatants antibodies against schistosome Mr 31 000 components. Electrophoretically separated total S. mansoni proteins were transferred onto nitrocellulose, and the position of the Mr 31 000 components was determined with polyclonal antisera and immunoblotting. Pieces of about 3 square mm of nitrocellulose bearing the diagnostic proteins were incubated with about 100 μ1 of hybridoma supernatant in microtitre plates and subsequently probed with peroxidase-conjugated antibody to mouse IgG. This screening technique identified hybridomas secreting antibody to the relevant S. mansoni antigens. It is applicable to other defined parasit antigens, which are, however, not available in biochemically purified form. The monoclonal antibodies produced against the proteins with diagnostic potential reacted with antigens localized in the schistosome gut.

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Bains, William. "Simplified format for DNA probe-based tests." Clinical Chemistry 37, no. 2 (February 1, 1991): 248–53. http://dx.doi.org/10.1093/clinchem/37.2.248.

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Abstract The approach I describe to using DNA probes in diagnostic tests is simpler than most existing formats. DNA in a sample is labeled by chemical reaction with bisulfite and methylamine to generate a sulfonated derivative. The DNA need not be purified to do this. The labeled sample is then incubated with an unlabeled, purified probe DNA, which is immobilized to a solid support. The amount of label remaining on the solid support after washing is detected by a monoclonal antibody that recognizes modified cytosines. The intensity of the signal depends on the amount of target DNA in the sample. Detection limits depend on the amount of immobilized DNA and on the degree of physical entrapment of the labeled DNA in sample material, but can be as low as 5 pg. This format is well suited to automation for use with existing robotic enzyme immunoassay procedures.

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Sapozhnikova, Ksenia A., Vsevolod A. Misyurin, Dmitry Y. Ryazantsev, Egor A. Kokin, Yulia P. Finashutina, Anastasiya V. Alexeeva, Igor A. Ivanov, et al. "Sensitive Immunofluorescent Detection of the PRAME Antigen Using a Practical Antibody Conjugation Approach." International Journal of Molecular Sciences 22, no. 23 (November 27, 2021): 12845. http://dx.doi.org/10.3390/ijms222312845.

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Bioconjugation of antibodies with various payloads has diverse applications across various fields, including drug delivery and targeted imaging techniques. Fluorescent immunoconjugates provide a promising tool for cancer diagnostics due to their high brightness, specificity, stability and target affinity. Fluorescent antibodies are widely used in flow cytometry for fast and sensitive identification and collection of cells expressing the target surface antigen. Nonetheless, current approaches to fluorescent labeling of antibodies most often use random modification, along with a few rather sophisticated site-specific techniques. The aim of our work was to develop a procedure for fluorescent labeling of immunoglobulin G via periodate oxidation of antibody glycans, followed by oxime ligation with fluorescent oxyamines. Here, we report a novel technique based on an in situ oxime ligation of ethoxyethylidene-protected aminooxy compounds with oxidized antibody glycans. The approach is suitable for easy modification of any immunoglobulin G, while ensuring that antigen-binding domains remain intact, thus revealing various possibilities for fluorescent probe design. The technique was used to label an antibody to PRAME, a cancer-testis protein overexpressed in a number of cancers. A 6H8 monoclonal antibody to the PRAME protein was directly modified with protected-oxyamine derivatives of fluorescein-type dyes (FAM, Alexa488, BDP-FL); the stoichiometry of the resulting conjugates was characterized spectroscopically. The immunofluorescent conjugates obtained were applied to the analysis of bone marrow samples from patients with oncohematological diseases and demonstrated high efficiency in flow cytometry quantification. The approach can be applied for the development of various immunofluorescent probes for detection of diagnostic and prognostic markers, which can be useful in anticancer therapy.

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Zhu, Lei, Hannu Koistinen, Ulf Landegren, and Ulf-Håkan Stenman. "Proximity Ligation Measurement of the Complex between Prostate Specific Antigen and α1-Protease Inhibitor." Clinical Chemistry 55, no. 9 (September 1, 2009): 1665–71. http://dx.doi.org/10.1373/clinchem.2009.127779.

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Abstract Background: Prostate specific antigen (PSA)–α1-protease inhibitor complex (PSA-API) is a minor form of PSA in serum. It may be useful for prostate cancer (PCa) diagnosis, but its specific detection is hampered by nonspecific background. To avoid this, we developed an immunoassay for PSA-API based on proximity ligation. Methods: We used a monoclonal antibody (mAb) to total PSA (tPSA) to capture PSA, while using another anti-tPSA mAb together with an anti-API mAb as probes. We measured PSA-API by quantification of amplified DNA strands conjugated to the probes. We measured serum PSA-API in 84 controls and 55 men with PCa who had PSA concentrations of 4.0–10 μg/L. Results: The detection limit of the assay was 6.6 ng/L. The proportion of PSA-API to tPSA (%PSA-API) tended to be lower in men with PCa (2.8%) than without cancer (3.3%) but was not statistically significant (P = 0.363). When used alone, %PSA-API [area under the curve (AUC) 0.546] did not improve detection of PCa, whereas %fPSA (AUC 0.710) and the sum of %fPSA and %PSA-API (AUC 0.723) did. At 90% diagnostic sensitivity, the diagnostic specificity for cancer was not significantly better for %fPSA + %PSA-API than for %fPSA alone (36% vs 30%). Conclusions: Proximity ligation eliminated nonspecific background, enabling accurate measurement of PSA-API in serum specimens with moderately increased tPSA. The combined use of %PSA-API and %fPSA provided a modest improvement for PCa detection, but based on the current study cohort, it is uncertain whether the improvement has clinical utility. .

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Lamy, A., E. Labourier, O. Prigneau, F. Di Fiore, R. Sesboüá, and J. Sabourin. "A method to assess KRAS/BRAF genotype in patients with metastatic colorectal cancer (mCRC) elligible for anti-EGFR therapies." Journal of Clinical Oncology 29, no. 4_suppl (February 1, 2011): 383. http://dx.doi.org/10.1200/jco.2011.29.4_suppl.383.

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383 Background: KRAS/BRAF mutation screening is a new diagnostic tool since activating mutations, especially for KRAS, are strong negative predictive factors for anti-EGFR monoclonal antibody therapies. Therefore, standardized, reliable, and rapid mutation detection methods are needed. The goal of the present study is to evaluate the concordance between a clinically validated laboratory-developed KRAS/BRAF SNaPshot test and a novel research use assay, the Signature KRAS/BRAF Mutations kit (Asuragen Inc.), in representative clinical specimens. Methods: Genomic DNA from FFPE mCRC specimens previously tested with the SNaPshot assay were analyzed using the Signature KRAS/BRAF mutations assay. The Signature assay consists of multiplex PCR followed by multiplex hybridization to fluorescent beads coupled to capture probes and detection on the Luminex analyzer. The qualitative assay detects 12 mutations in KRAS codons 12/13 and BRAF V600E based on the median fluorescence intensity (MFI) generated by each bead above (positive) or below (negative) a fixed cut off. Results: To date, KRAS/BRAF mutational status was successfully assessed with the signature assay in 248 samples. After initial testing, signature and SNaPshot assays were in qualitative agreement for 232 samples (93.5%): 166 positive for KRAS, 46 positive for BRAF, and 20 double negative. 16 samples (6%) presented discrepant results: (i) 11 false negative samples with fluorescence signals for the expected mutations above the average assay background signal but below the 450 MFI positive cut off preselected for this study (330-431 MFI), (ii) 3 samples positive for both KRAS and BRAF and (iii) 2 false positive samples. 7 discrepant samples have been retested to date resulting in an overall agreement of 99.6% (239/240). However, different KRAS mutations were identified by the SNaPshot and signature assays in 3 positive samples. Conclusions: The signature KRAS/BRAF mutations assay is an attractive method, easy to use, less time consuming than the SNaPshot assay, and potentially adaptable to routine clinical testing. Further analysis will establish and validate cut offs to be used in routine diagnostic procedures. No significant financial relationships to disclose.

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9

Dunn, Bruce E. "Nucleic acid and monoclonal antibody probes, applications in diagnostic microbiology." Human Pathology 22, no. 4 (April 1991): 403–4. http://dx.doi.org/10.1016/0046-8177(91)90095-7.

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Parratt, D. "Nucleic Acid and Monoclonal Antibody Probes. Applications in Diagnostic Microbiology." Journal of Clinical Pathology 42, no. 12 (December 1, 1989): 1313. http://dx.doi.org/10.1136/jcp.42.12.1313-c.

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Paterson, Marion, and John F. Kennedy. "Nucleic Acid and Monoclonal Antibody Probes. Applications in diagnostic microbiology." Carbohydrate Polymers 18, no. 1 (January 1992): 73–74. http://dx.doi.org/10.1016/0144-8617(92)90191-r.

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12

Săndulescu, Oana, Ioana Viziteu, Anca Streinu-Cercel, Victor Daniel Miron, Liliana Lucia Preoțescu, Narcis Chirca, Simona Elena Albu, Mihai Craiu, and Adrian Streinu-Cercel. "Novel Antimicrobials, Drug Delivery Systems and Antivirulence Targets in the Pipeline—From Bench to Bedside." Applied Sciences 12, no. 22 (November 16, 2022): 11615. http://dx.doi.org/10.3390/app122211615.

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In a fast-paced medical reality, biosciences and bioengineering have become essential components in medical research and development. The aim of this paper is to characterize the recent progresses made in fighting antimicrobial resistance, particularly in relation to WHO’s priority pathogens, by providing an in-depth review of novel antimicrobials, drug delivery systems for targeted antimicrobial action and novel antivirulence targets. We systematically searched the ClinicalTrials.gov database to identify clinical trials targeting WHO’s priority 1 (critical) pathogens: carbapenem-resistant Acinetobacter baumannii, carbapenem-resistant Pseudomonas aeruginosa, and carbapenem-resistant ESBL-producing Enterobacteriaceae. We identified a limited number of clinical trials, specifically for: one novel betalactamase inhibitor for Acinetobacter spp., one anti-virulence human monoclonal antibody for Pseudomonas spp. and no novel antimicrobials for carbapenem-resistant Enterobacteriaceae. We also performed a review of field literature to exemplify the main applications of drug delivery systems in infectious diseases, particularly in achieving targeted antibiotic distribution, in enhancing local activity with reduced off-target effects, triggered antibiotic release and triggered antibacterial photodynamic therapy. We conclude by presenting novel targets for antivirulence therapeutics that act by disrupting quorum sensing, inhibiting bacterial adherence and biofilm formation, silencing virulence traits and neutralizing bacterial toxins. Furthermore, the main principles of rational antimicrobial use are highlighted, in an effort to describe potential areas for targeted intervention, from diagnostic stewardship to antimicrobial stewardship.

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Shults, Keith, David R. Head, Leanne Flye, Madan H. Jagasia, Stacey A. Goodman, Sara A. McClintock-Treep, Fouad I. Boulos, James W. Jacobberger, Greg Stelzer, and Robert C. Briggs. "Myeloid Nuclear Differentiation Antigen (MNDA) Protein Expression in Myelodysplastic Syndrome (MDS) Is Reduced in Maturing Myeloid Cells." Blood 104, no. 11 (November 16, 2004): 4734. http://dx.doi.org/10.1182/blood.v104.11.4734.4734.

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Abstract Expression of the human MNDA protein is restricted to hematopoietic cells, and occurs at uniformly high level only in the normal granulocyte/monocyte lineage. Myeloblasts and all later stages of the lineage express MNDA, while stem cells do not (Hum Pathol1999;30:1040). Gene expression analyses of MDS cases have shown that MNDA mRNA is significantly down regulated in familial MDS and in high-risk sporadic MDS cases (Blood2000;100:3553 and Leukemia2004;18:449). We evaluated MNDA protein expression in patients with MDS. Methods: We directly conjugated Alexa 488 (Molecular Probes) to a rat monoclonal antibody specific for MNDA for use in multiparameter cytometric analysis with CD45/CD34 and DRAQ5 (DNA content). Patients’ (n=17) clinical histories, laboratory data, cytogenetics, and morphology were reviewed for confirmation of diagnoses and blinded to study results. Patients lacking primary marrow disease (n=9) were used as controls. Analysis was performed using Winlist 5.0 software (Verity Software) with DDE links to ModFitLT 3.0 using modifications of published methods along with Esoterix Center for Innovation generated algorithms. Analyses were blinded to clinical results. Results: The normal dataset revealed MNDA expression in expected cell populations, with only a small subset of CD34+ cells exhibiting MNDA (7.5% on average). Expression of MNDA protein in maturing myeloid forms in normals was consistently greater than in MDS samples (MFI=802 vs. 569 in MDS; MFI=mean fluorescence intensity defined by the linear mean of each population). Six of 17 MDS cases displayed greater than 10% maturing myeloid forms lacking MNDA protein expression (never observed in normals). The % MNDA+/CD34+ cells was identical in the two datasets (7.5% in normal vs. 7.6% in MDS). Discussion: Our results suggest that MNDA protein expression in MDS patients follows a pattern similar to that in gene expression analyses. We additionally demonstrate that MNDA protein expression at the cellular level is variable in MDS, versus uniform expression in normals; parallel data for MNDA message expression at the cellular level is not available. The finding of MNDA-negative myeloid forms in MDS may have diagnostic utility, and may provide insight into both impaired myeloid differentiation in MDS and the biologic role of MNDA.

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Woo, Patrick T. K. "The use of monoclonal antibody probes in salmonid cryptobiosis." Aquaculture 177, no. 1-4 (July 1999): 311–24. http://dx.doi.org/10.1016/s0044-8486(99)00094-0.

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ITOH, KUNIHIKO, SHUNJI ISHIWATA, NAKAO ISHIDA, and MICHINAO MIZUGAKI. "Diagnostic Use of Anti-Modified Nucleoside Monoclonal Antibody." Tohoku Journal of Experimental Medicine 168, no. 2 (1992): 329–31. http://dx.doi.org/10.1620/tjem.168.329.

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Liu, Hung J., Joseph J. Giambrone, Yeong H. Wu, Ming H. Liao, and Che F. Lu. "The use of monoclonal antibody probes for the detection of avian reovirus antigens." Journal of Virological Methods 86, no. 2 (May 2000): 115–19. http://dx.doi.org/10.1016/s0166-0934(00)00137-3.

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Leong, M. M., C. Milstein, and R. Pannell. "Luminescent detection method for immunodot, Western, and Southern blots." Journal of Histochemistry & Cytochemistry 34, no. 12 (December 1986): 1645–50. http://dx.doi.org/10.1177/34.12.3537113.

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An anti-peroxidase-anti-biotin hybrid hybridoma rat cell line, capable of producing a bispecific monoclonal antibody, has been derived to explore its use in conjunction with a luminol immunodetection system. Luminescence was detected using x-ray film. The method was sufficiently sensitive and effective, but was less sensitive than autoradiographic methods using high-specific-activity 32P-labeled probes. Exposure times, on the other hand, were of the order of seconds rather than days. The direct binding of both peroxidase and biotin by the bispecific monoclonal antibody is simpler but less sensitive than the more conventional indirect method using a commercial peroxidase coupled with anti-rat antibody as a developing antibody.

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Reddy, M. V. R., K. A. Parkhe, B. C. Harinath, and W. F. Piessens. "Wb e34 monoclonal antibody: Further characterization and diagnostic use in bancroftian filariasis." Journal of Clinical Laboratory Analysis 3, no. 5 (1989): 277–81. http://dx.doi.org/10.1002/jcla.1860030504.

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Lee, Long Huw. "The use of monoclonal antibody probes for the detection of infectious bursal disease virus antigens." Avian Pathology 21, no. 1 (January 1992): 87–96. http://dx.doi.org/10.1080/03079459208418821.

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Etrych, Tomas, Alena Braunova, David Zogala, Lukas Lambert, Nicol Renesova, and Pavel Klener. "Targeted Drug Delivery and Theranostic Strategies in Malignant Lymphomas." Cancers 14, no. 3 (January 26, 2022): 626. http://dx.doi.org/10.3390/cancers14030626.

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Malignant lymphomas represent the most common type of hematologic malignancies. The first clinically approved TDD modalities in lymphoma patients were anti-CD20 radioimmunoconjugates (RIT) 131I-tositumomab and 90Y-ibritumomab-tiuxetan. The later clinical success of the first approved antibody–drug conjugate (ADC) for the treatment of lymphomas, anti-CD30 brentuximab vedotin, paved the path for the preclinical development and clinical testing of several other ADCs, including polatuzumab vedotin and loncastuximab tesirine. Other modalities of TDD are based on new formulations of “old” cytostatic agents and their passive trapping in the lymphoma tissue by means of the enhanced permeability and retention (EPR) effect. Currently, the diagnostic and restaging procedures in aggressive lymphomas are based on nuclear imaging, namely PET. A theranostic approach that combines diagnostic or restaging lymphoma imaging with targeted treatment represents an appealing innovative strategy in personalized medicine. The future of theranostics will require not only the capability to provide suitable disease-specific molecular probes but also expertise on big data processing and evaluation. Here, we review the concept of targeted drug delivery in malignant lymphomas from RIT and ADC to a wide array of passively and actively targeted nano-sized investigational agents. We also discuss the future of molecular imaging with special focus on monoclonal antibody-based and monoclonal antibody-derived theranostic strategies.

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Frosch, M., W. Peuckert, and D. Bitter-Suermann. "Diagnostic use of monoclonal IgG antibody to meningococcal B polysaccharide in cerebrospinal fluid." Antonie van Leeuwenhoek 52, no. 3 (1986): 253–54. http://dx.doi.org/10.1007/bf00555250.

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Parks, WM, RD Gingrich, CE Dahle, and JC Hoak. "Identification and characterization of an endothelial, cell-specific antigen with a monoclonal antibody." Blood 66, no. 4 (October 1, 1985): 816–23. http://dx.doi.org/10.1182/blood.v66.4.816.816.

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Abstract The purpose of these studies was to use monoclonal antibodies to identify and characterize plasma membrane components unique to the vascular endothelium. Our assumption is that such components may perform some of the specialized functions of the endothelium and, by their identification with antibody probes, we may be able to study further their function and structure. Thus, primary cultures of human umbilical vein endothelium were used to immunize mice whose spleen cells were fused with the mouse myeloma cell NS-1. HEC-1 is a monoclonal antibody derived from such a fusion that appears to react with an antigen located only on endothelial cells. The antigen has been characterized by immunoprecipitation and polyacrylamide gel electrophoresis as a glycoprotein with a mol wt of 180,000 daltons under nonreducing conditions and 90,000 daltons under reducing conditions. Despite a close resemblance to a membrane component shown by others to be a receptor for transferrin, several lines of evidence reported in this paper indicate that this is not the function of the HEC-1 antigen. These data show that monoclonal antibodies can be used to identify and characterize membrane components of the vascular endothelium. Moreover, these probes can be used to inquire about the structure and function of the antigen with which they react.

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Parks, WM, RD Gingrich, CE Dahle, and JC Hoak. "Identification and characterization of an endothelial, cell-specific antigen with a monoclonal antibody." Blood 66, no. 4 (October 1, 1985): 816–23. http://dx.doi.org/10.1182/blood.v66.4.816.bloodjournal664816.

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The purpose of these studies was to use monoclonal antibodies to identify and characterize plasma membrane components unique to the vascular endothelium. Our assumption is that such components may perform some of the specialized functions of the endothelium and, by their identification with antibody probes, we may be able to study further their function and structure. Thus, primary cultures of human umbilical vein endothelium were used to immunize mice whose spleen cells were fused with the mouse myeloma cell NS-1. HEC-1 is a monoclonal antibody derived from such a fusion that appears to react with an antigen located only on endothelial cells. The antigen has been characterized by immunoprecipitation and polyacrylamide gel electrophoresis as a glycoprotein with a mol wt of 180,000 daltons under nonreducing conditions and 90,000 daltons under reducing conditions. Despite a close resemblance to a membrane component shown by others to be a receptor for transferrin, several lines of evidence reported in this paper indicate that this is not the function of the HEC-1 antigen. These data show that monoclonal antibodies can be used to identify and characterize membrane components of the vascular endothelium. Moreover, these probes can be used to inquire about the structure and function of the antigen with which they react.

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Gottlin, Elizabeth B., Xiangrong Guan, Charles Pegram, Allen Cannedy, Michael J. Campa, and Edward F. Patz. "Isolation of Novel EGFR-Specific VHH Domains." Journal of Biomolecular Screening 14, no. 1 (November 21, 2008): 77–85. http://dx.doi.org/10.1177/1087057108327064.

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Epidermal growth factor receptor (EGFR) is overexpressed or mutated in a high percentage of tumors. EGFR has long been considered a promising target for cancer diagnostic and therapeutic applications. However, monoclonal antibodies and other large antibody constructs diffuse into tumors slowly, limiting their efficacy. To develop lower molecular weight probes for EGFR and other tumor cell receptors, the authors immunized a llama with the extracellular domains (ECDs) of EGFR and an oncogenic mutant receptor, EGFRvIII, and with extracts of tumor cell lines. From the immune repertoire of the llama, the authors constructed a heavy chain variable domain (VHH domain)—phage library. At ~16 kDa, the VHH domain is a tenth of the size of a monoclonal antibody and is the smallest antibody fragment that retains specificity. By affinity selection from this library, the authors isolated many VHH domains with specificity for EGFR. The VHH domains bind to whole cells expressing the receptor but not to control cells lacking the receptor and can immunoprecipitate EGFR from cell lysates. Some VHH domains have cross-specificity with existing anti-EGFR monoclonal antibodies and have reasonably high (nM) affinities. The llama-VHH domain library is also potentially a rich source of targeting agents directed toward other tumor cell receptors. ( Journal of Biomolecular Screening 2009:77-85)

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Barnard, R., P. G. Bundesen, D. B. Rylatt, and M. J. Waters. "Evidence from the use of monoclonal antibody probes for structural heterogeneity of the growth hormone receptor." Biochemical Journal 231, no. 2 (October 15, 1985): 459–68. http://dx.doi.org/10.1042/bj2310459.

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We describe the use of four monoclonal antibodies (MAbs) to the rabbit liver growth hormone (GH) receptor and one raised against purified rat liver GH receptor to characterize liver receptor subtypes which differ in their hormone-binding regions. The anti-(rat liver GH receptor) MAb both inhibited and precipitated rat and rabbit GH receptors, but only one-half of 125I-oGH (ovine GH) binding to liver microsomes could be inhibited by excess antibody. Conversely, only one-half of 125I-anti-(rat GH receptor) MAb binding was inhibited by excess oGH and Scatchard plots for this MAb exhibited two components. Although only 50% of 125I-oGH binding to membranes was inhibited by this MAb, all solubilized receptor could be immunoprecipitated. We postulate two epitopes for the anti-(rat GH receptor) MAb, one located at the hormone-binding site (inhibitory site) and one elsewhere (immunoprecipitating site). A second, rabbit-specific antibody (MAb 7) inhibited 85% of hormone binding but only 30% of 125I-anti-(rat GH receptor) MAb binding to rabbit liver microsomes. A combination of this MAb with the anti-(rat GH receptor) MAb totally inhibited 125I-oGH binding. MAb 7 alone totally inhibited 125I-rat GH binding to rabbit liver microsomes, as it did with 125I-oGH binding to purified receptor. On the basis of these results and others we postulate three types of GH receptor in rabbit liver membranes and ascribe approximate extents of 125I-oGH binding to each. A cytosolic ‘GH receptor’ which is not poly(ethylene glycol)-precipitable is shown to share five epitopes with ‘type 2’ microsomal receptors. Purified plasma membrane and endoplasmic reticulum fractions derived from a rabbit liver microsomal preparation have identical antigenic characteristics with respect to the GH-binding region, indicating that the heterogeneity we describe is not related to receptor processing. Of the three types of GH receptor in the plasma membrane of the rabbit (and possibly rat) we postulate that one (type 1) corresponds to the GH receptor involved in stimulating growth and possesses all of the epitopes studied here. A second (type 2) appears to be identical with the cytosolic ‘GH receptor’ and lacks the epitope for the anti-(rat GH receptor) MAb in the hormone binding site region. A third (type 3) does not possess the epitope for the inhibitory anti-(rabbit GH receptor) MAb, appears not to bind rat GH and is lost during purification. The availability of type-specific MAbs will facilitate assignment of specific functions to liver receptor subtypes which mediate the multiple functions of GH.

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Kristensen, Josrgen Schosler, and Peter Hokland. "Monoclonal antibody ratios in malignant myeloid diseases: diagnostic and prognostic use in myelodysplastic syndromes." British Journal of Haematology 74, no. 3 (March 1990): 270–76. http://dx.doi.org/10.1111/j.1365-2141.1990.tb02582.x.

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Speare, D. J., R. J. F. Markham, B. Despres, K. Whitman, and N. MacNair. "Examination of Gills from Salmonids with Bacterial Gill Disease using Monoclonal Antibody Probes for Flavobacterium Branchiophilum and Cytophaga Columnaris." Journal of Veterinary Diagnostic Investigation 7, no. 4 (October 1995): 500–505. http://dx.doi.org/10.1177/104063879500700413.

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Bacterial diseases of the gills of commercially reared salmonids in freshwater are common problems. They accounted for 18% of all diagnostic submissions to the Atlantic Veterinary College from commercial fish hatcheries. Definitive diagnosis is difficult because of the growth characteristics of the putative bacteria in culture. Research into the pathogenesis of these diseases has also been similarly limited. Monoclonal antibodies (MAbs) were developed to 2 globally significant gill bacterial pathogens, Flavobacterium branchiophilum, the causative agent of bacterial gill disease, and Cytophaga columnaris, the causative agent of columnar-is disease of salmonids. These MAbs were then used as the basis for an indirect fluorescent antibody test to assess archived cases of gill disease in our region. Flavobacterium branchiophilum was the dominant bacterium detected in the biofilm of diseased gills in our study region. Of the cases tentatively diagnosed based on histopathology as bacterial gill disease, 76.2% tested positively with the MAbs to F. branchiophilum. Also present within 18.7% of these cases were bacteria which reacted positively to the MAbs for C. columnaris. We conclude that the MAbs produced are valuable diagnostic and research probes for common bacterial diseases of the gills of salmon and trout in Atlantic Canada. This study also adds further proof that F. branchiophilum acting alone can be sufficient cause for bacterial gill disease.

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Sanson, Bernd-Jan, Wouter de Monyé, Jeroen Lijmer, Menno Huisman, Harry Büller, Willem Nieuwenhuizen, Dees P. Brandjes, and Melvin Mac Gillavry. "Use of a New Monoclonal Antibody-Based Enzyme Immunoassay for Soluble Fibrin to Exclude Pulmonary Embolism." Thrombosis and Haemostasis 84, no. 09 (2000): 474–77. http://dx.doi.org/10.1055/s-0037-1614047.

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SummaryWe prospectively evaluated the diagnostic performance of a new soluble fibrin assay in 303 consecutive patients with suspected pulmonary embolism and examined potentially useful cut-off levels at which this disease can be safely excluded. In addition, the diagnostic accuracy was calculated in the subgroups of in- and outpatients. The ROC curve of the assay in the total study cohort had an area under the curve of 0.69. The cut-off level associated with a sensitivity and negative predictive value of 100% was 20 ng/ml, but the specificity was only 4%. The cut-off level with a sensitivity of 90% was 30 ng/ml, which corresponded with a specificity and negative predictive value of 27% and 86%, respectively. The diagnostic performance was comparable in the subgroups of in- and outpatients. We conclude that the soluble fibrin assay has a low diagnostic accuracy and seems unsuitable as a screening test for the exclusion of pulmonary embolism.

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Parker, Tracy Dewese, Noritoshi Kitamoto, Tomoyuki Tanaka, Anne M. Hutson, and Mary K. Estes. "Identification of Genogroup I and Genogroup II Broadly Reactive Epitopes on the Norovirus Capsid." Journal of Virology 79, no. 12 (June 15, 2005): 7402–9. http://dx.doi.org/10.1128/jvi.79.12.7402-7409.2005.

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ABSTRACT Norwalk virus, a member of the family Caliciviridae, is an important cause of acute epidemic nonbacterial gastroenteritis. Norwalk and related viruses are classified in a separate genus of Caliciviridae called Norovirus, which is comprised of at least three genogroups based on sequence differences. Many of the currently available immunologic reagents used to study these viruses are type specific, which limits the identification of antigenically distinct viruses in detection assays. Identification of type-specific and cross-reactive epitopes is essential for designing broadly cross-reactive diagnostic assays and dissecting the immune response to calicivirus infection. To address this, we have mapped the epitopes on the norovirus capsid protein for both a genogroup I-cross-reactive monoclonal antibody and a genogroup II-cross-reactive monoclonal antibody by use of norovirus deletion and point mutants. The epitopes for both monoclonal antibodies mapped to the C-terminal P1 subdomain of the capsid protein. Although the genogroup I-cross-reactive monoclonal antibody was previously believed to recognize a linear epitope, our results indicate that a conformational component of the epitope explains the monoclonal antibody's genogroup specificity. Identification of the epitopes for these monoclonal antibodies is of significance, as they are components in a commercially available norovirus-diagnostic enzyme-linked immunosorbent assay.

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Brigati, Jennifer, David D. Williams, Iryna B. Sorokulova, Viswaprakash Nanduri, I.-Hsuan Chen, Charles L. Turnbough, and Valery A. Petrenko. "Diagnostic Probes for Bacillus anthracis Spores Selected from a Landscape Phage Library." Clinical Chemistry 50, no. 10 (October 1, 2004): 1899–906. http://dx.doi.org/10.1373/clinchem.2004.038018.

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AbstractBackground: Recent use of Bacillus anthracis spores as a bioweapon has highlighted the need for a continuous monitoring system. Current monitoring systems rely on antibody-derived probes, which are not hardy enough to withstand long-term use under extreme conditions. We describe new, phage-derived probes that can be used as robust substitutes for antibodies.Methods: From a landscape phage library with random octapeptides displayed on all copies of the major phage coat protein of the phage fd-tet, we selected clones that bound to spores of B. anthracis (Sterne strain). ELISA, micropanning, and coprecipitation assays were used to evaluate the specificity and selectivity with which these phage bound to B. anthracis spores.Results: Peptides on the selected clones directed binding of the phage to B. anthracis spores. Most clones exhibited little or no binding to spores of distantly related Bacillus species, but some binding was observed with spores of closely related species. Our most specific spore-binding phage displayed a peptide EPRLSPHS (several thousand peptides per phage) and bound 3.5- to 70-fold better to spores of B. anthracis Sterne than to spores of other Bacillus species.Conclusions: The selected phage probes bound preferentially to B. anthracis Sterne spores compared with other Bacillus species. These phage could possibly be further developed into highly specific and robust probes suitable for long-term use in continuous monitoring devices and biosorbents.

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Chico, Martha E., Ronald H. Guderian, Philip J. Cooper, Rodrigo Armijos, and Max Grogl. "Evaluation of a direct immunofluorescent antibody (difma) test using Leishmania genus - specific monoclonal antibody in the routine diagnosis of cutaneous leishmaniasis." Revista da Sociedade Brasileira de Medicina Tropical 28, no. 2 (June 1995): 99–103. http://dx.doi.org/10.1590/s0037-86821995000200002.

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A direct immunofluorescent antibody (DIFMA) test using a Leishmania genus- specific monoclonal antibody was evaluated in the routine diagnosis of cutaneous leishmaniasis (CL) in Ecuador. This test was compared with the standard diagnostic techniques of scrapings, culture and histology. Diagnostic samples were taken from a total of 90 active dermal ulcers from patients from areas of Ecuador known to be endemic for cutaneous leishmaniasis. DIFMA was positive in all lesions. It was shown to be significantly superior to standard diagnostic methods either alone or in combination. The sensitivity of DIFMA did not diminish with chronicity of lesions. This test proved to be extremely useful in the routine diagnosis of CL because it is highly sensitive, is easy to use and produces rapid results.

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Kumar, N. Shashi, Narayan Moger, Chidanand A. Rabinal, P. U. Krishnaraj, and K. N. Chandrashekara. "Production of Diagnostic Kit to Detect Cry 2B Antigen by Use of scFv Monoclonal Antibody." International Journal of Current Microbiology and Applied Sciences 6, no. 7 (July 10, 2017): 4401–11. http://dx.doi.org/10.20546/ijcmas.2017.607.459.

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Tachibana, Hiroshi, Masataka Takekoshi, Xun-Jia Cheng, Yuta Nakata, Tsutomu Takeuchi, and Seiji Ihara. "Bacterial Expression of a Human Monoclonal Antibody-Alkaline Phosphatase Conjugate Specific for Entamoeba histolytica." Clinical Diagnostic Laboratory Immunology 11, no. 1 (January 2004): 216–18. http://dx.doi.org/10.1128/cdli.11.1.216-218.2004.

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ABSTRACT We previously produced human monoclonal antibody Fab fragments specific to Entamoeba histolytica in Escherichia coli. In order to use these Fab fragments for diagnostic purposes, an expression vector to produce a fusion protein of Fab and alkaline phosphatase (PhoA) in E. coli was designed and constructed. The E. coli PhoA gene was fused to the 3′ terminus of the gene encoding the heavy-chain Fd region. The kappa and Fd genes from a previously prepared antibody clone, CP33, which is specific for the 260-kDa lectin of E. histolytica, were used as human antibody genes. When the fusion protein of CP33 and PhoA was incubated with paraformaldehyde-fixed trophozoites of E. histolytica and developed with a substrate, the trophozoites appeared to be stained. These results demonstrate the feasibility of bacterial expression of a human monoclonal antibody-PhoA conjugate specific for E. histolytica and that the antibody can be used to detect E. histolytica antigen without the use of chemically conjugated secondary antibodies.

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Adams, Alexandra, Kimberly D. Thompson, David Morris, Carlos Farias, and Shi Chu Chen. "Development and use of monoclonal antibody probes forimmunohistochemistry, ELISA and IFAT to detect bacterial and parasitic fish pathogens." Fish & Shellfish Immunology 5, no. 8 (November 1995): 537–47. http://dx.doi.org/10.1016/s1050-4648(95)80040-9.

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Smit, L. M., H. Veldman, and F. G. Jennekens. "Immunohistochemical localization of acetylcholine receptors at human endplates using a monoclonal antibody." Journal of Histochemistry & Cytochemistry 35, no. 5 (May 1987): 613–17. http://dx.doi.org/10.1177/35.5.3549892.

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We describe a simple indirect immunohistochemical method for localization of acetylcholine receptors (AChR) in motor endplates at the light and electron microscopic level. This method involves the use of a monoclonal antibody directed against the main immunogenic region (MIR) of AChRs and is applicable to periodate-lysine-paraformaldehyde (PLP)-fixed tissue. We discuss the advantages of this method, as compared with the alpha-bungarotoxin-immunoperoxidase technique, and stress its value for diagnostic investigations of motor point biopsies from patients with neuromuscular transmission disorders.

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Wessel, G. M., and D. R. McClay. "Two embryonic, tissue-specific molecules identified by a double-label immunofluorescence technique for monoclonal antibodies." Journal of Histochemistry & Cytochemistry 34, no. 6 (June 1986): 703–6. http://dx.doi.org/10.1177/34.6.3084626.

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We identify two tissue-specific molecules in the sea urchin embryo by an immunofluorescence technique capable of co-localizing monoclonal antibodies on the same tissue section. The technique uses monovalent Fab-fluorochrome conjugates as secondary reagents to avoid cross-talk of subsequent antibody probes. Using this technique, we show that two cell surface molecules are expressed by different cell populations in the embryo. The technique is generally applicable for antibodies regardless of species or subtype specificity, and uses commercially available reagents. The technique provides sufficient amplification and resolution for analytical work, yet is rapid enough for screening procedures. As a fluorescent counterstain, use of the dye 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS) in the protocol provides a distinct fluorescent background staining of the tissue without interference with the specific antibody staining.

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Clemo, F. A. S., D. B. DeNicola, and L. J. Delaney. "Immunohistochemical Evaluation of Canine Carcinomas with Monoclonal Antibody B72.3." Veterinary Pathology 30, no. 2 (March 1993): 140–45. http://dx.doi.org/10.1177/030098589303000206.

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Monoclonal antibody (MAb) B72.3 reacts with a tumor-associated glycoprotein, designated TAG-72. TAG-72 has been identified in many human carcinomas but generally is not found in normal human tissues. Because of its proven utility in the diagnosis of human carcinomas, MAb B72.3 was applied to several different types of canine carcinomas. Five types of formalin-fixed, paraffin-embedded canine carcinomas were evaluated for immunoreactivity with MAb B72.3 by use of an avidin biotin immunoperoxidase complex method. Samples were considered positive when ≥5% of all malignant cells contained a distinct intracellular stain. Immunoreactivity for MAb B72.3 was observed in 6/9 (67%) pulmonary adenocarcinomas, 7/13 (54%) transitional cell carcinomas, 7/11 (64%) mammary adenocarcinomas, 7/11 (64%) nasal adenocarcinomas, and 1/2 (50%) prostatic adenocarcinomas. The average cellular staining for positive carcinomas was 25%. Normal canine tissues from similar anatomic sites had little or no individual cell immunoreactivity. These preliminary results indicate that some canine carcinomas may express a tumor-associated antigen that is similar to TAG-72 and that MAb B72.3 immunoreactivity may be of diagnostic significance in classifying animal tumors.

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Loreto, Carla, and Rosario Caltabiano. "Immunohistochemical Expression." Applied Sciences 11, no. 1 (January 1, 2021): 360. http://dx.doi.org/10.3390/app11010360.

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Immunohistochemistry (IHC) is an ancillary method, widely used in pathologist practice, that allows to identify diagnostic and prognostic/predictive therapeutic response protein markers on tissue samples by the use of specific monoclonal antibodies and chromogenic substances that guarantee the visualization of the antibody–antigene binding complex under the light microscope [...]

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Cimaglia, Fabio, Alessandro Aliverti, Maurizio Chiesa, Palmiro Poltronieri, Enrico De Lorenzis, Angelo Santino, and Leonardo A. Sechi. "Quantum dots nanoparticle-based lateral flow assay for rapid detection of Mycobacterium species using anti-FprA antibodies." Nanotechnology Development 2, no. 1 (January 4, 2012): 5. http://dx.doi.org/10.4081/nd.2012.e5.

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A lateral flow (LF) device combined with quantum dots (QDs) technology was developed for rapid detection of a specific mycobacterial flavoprotein reductase (fprA). In order to develop the LF assay based on a double-antibody sandwich format, two monoclonal antibodies recognizing different epitopes located in separated fprA domains were identified. The first monoclonal antibody was immobilized onto the detection zone of a porous nitrocellulose membrane, whereas another monoclonal antibody was conjugated to QDs nanoparticles as a detection system. Using these monoclonal antibodies we recorded a good fluorescence signal, the intensity of which was directly proportional to the concentration of fprA protein. The use of antibodies conjugated with fluorescent semiconductor QDs via biotin-streptavidin bridge, allowed the detection of fprA protein at concentrations as low as 12.5 pg/μL in less than 10 min. The reported technology could be useful in the diagnostic investigation of Mycobacterium tuberculosis and other human pathogens in clinical specimens.

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Muhlen, Constantin von zur, Dominik von Elverfeldt, Karlheinz Peter, and Christoph Hagemeyer. "Single-chain antibodies as diagnostic tools and therapeutic agents." Thrombosis and Haemostasis 101, no. 06 (2009): 1012–19. http://dx.doi.org/10.1160/th08-12-0816.

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SummaryOver three decades after the generation of the first mouse monoclonal antibodies by Kohler and Milstein, recombinant antibodies are the fastest growing class of therapeutic proteins. Furthermore, antibodies are key detection reagents in research and diagnostics. Technology improvements have provided several approaches to manufacturing human antibodies with high affinity for biologically relevant targets. Approximately 300 development programs for therapeutic antibodies have been reported in industrial and academic laboratories, and this clearly demonstrates the expectations towards antibody technology. Antibody fragments are a subclass with growing clinical importance. This review focuses on single-chain antibodies as one of the smallest possible format for recombinant antibodies and their use as diagnostic tools and therapeutic agents. We describe the structure, selection and production of single-chain antibodies. Furthermore, we review current applications of antibody fragments focusing on thrombus targeting using fibrin- and platelet-specific single-chain antibodies as well as describing novel noninvasive imaging approaches for the diagnosis of thrombosis and inflammation.

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Pinkston, Kenneth L., Kavindra V. Singh, Peng Gao, Nathaniel Wilganowski, Holly Robinson, Sukhen Ghosh, Ali Azhdarinia, Eva M. Sevick-Muraca, Barbara E. Murray, and Barrett R. Harvey. "Targeting Pili in Enterococcal Pathogenesis." Infection and Immunity 82, no. 4 (January 22, 2014): 1540–47. http://dx.doi.org/10.1128/iai.01403-13.

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ABSTRACTPassive protection, the administration of antibodies to prevent infection, has garnered significant interest in recent years as a potential prophylactic countermeasure to decrease the prevalence of hospital-acquired infections. Pili, polymerized protein structures covalently anchored to the peptidoglycan wall of many Gram-positive pathogens, are ideal targets for antibody intervention, given their importance in establishing infection and their accessibility to antibody interactions. In this work, we demonstrated that a monoclonal antibody to the major component ofEnterococcus faecalispili, EbpC, labels polymerized pilus structures, diminishes biofilm formation, and significantly prevents the establishment of a rat endocarditis infection. The effectiveness of this anti-EbpC monoclonal provides strong evidence in support of its potential as a preventative. In addition, after radiolabeling, this monoclonal identified the site of enterococcal infection, providing a rare example of molecularly specific imaging of an established bacterial infection and demonstrating the versatility of this agent for use in future diagnostic and therapeutic applications.

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Saleh, M. N., D. M. Miller, L. Peterson, C. D. Russell, M. W. Unger, M. M. Urist, R. H. Wheeler, and A. F. LoBuglio. "Clinical use of a standard kit-preparation of radiolabeled monoclonal antibody 96.5 in the diagnostic imaging of metastatic melanoma." Journal of Clinical Oncology 6, no. 6 (June 1988): 1059–65. http://dx.doi.org/10.1200/jco.1988.6.6.1059.

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We studied the efficiency of a standard-kit preparation using 1 mg 111In-labeled 96.5 monoclonal antibody in combination with 19 mg of unlabeled antibody in the diagnostic imaging of 27 patients with documented metastatic melanoma. Twenty-three of 26 patients (88%) demonstrated immunoscintigraphic localization of tumor. Of 104 metastatic sites previously documented by conventional studies, 62 (60%) were identified by immunoscintigraphy. A total of 77 sites demonstrated localization of radiolabeled antibody. Fifty-four (70%) corresponded to known sites of disease; eight sites (10%) were "discovered" by immunoscintigraphy and subsequently confirmed by conventional studies; 15 imaged sites (20%) could not be confirmed by conventional studies. Size and location of metastasis appear to be important features that influence imaging efficiency. Tumor size (greater than or equal to 2 cm v less than 2 cm) appears to be the statistical dominant determinant. The feasibility and potential clinical use of radioimmune imaging of tumors is discussed.

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Kretenchuk, O. F. "Domestically-Produced Monoclonal-Antibody-Based Means of Diagnosing Particularly Dangerous Infections." Problems of Particularly Dangerous Infections, no. 4 (January 25, 2022): 35–45. http://dx.doi.org/10.21055/0370-1069-2021-4-35-45.

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Currently, great significance is attached to the preparation of diagnostic drugs based on specific immunoreagents, which include monoclonal antibodies produced by hybridomas. The use of monoclonal antibodies is one of the important approaches for the detection of pathogens of particularly dangerous infections – anthrax, brucellosis, tularemia, plague, cholera, glanders, and melioidosis. The review presents the main achievements of Russian scientists on obtaining such experimental drugs, and also pays attention to those sets of monoclonal reagents that are authorized in the Russian Federation. To date, three sets of reagents for detecting the causative agent of anthrax (latex agglutination, immunochromatographic method, multiplex immunofluorescence analysis) have been registered in our country on the basis of monoclonal antibodies; four sets of reagents for identifying the causative agent of tularemia (latex agglutination, immunochromatographic method, multiplex immunofluorescence analysis, dot-variant of enzyme immunoassay); three sets for the detection of plague microbe (enzyme immunoassay and immune chromatographic tests); five sets for cholera vibrios (slide agglutination, immunofluorescence, immune chromatographic method and enzyme immunoassay); two sets for the diagnosis of glanders and melioidosis (immunofluorescence); kits for detecting brucella have not been registered, there are only singular experimental designs. The involvement of modern drugs based on monoclonal antibodies in the diagnosis of particularly dangerous infections will improve the quality and reliability of laboratory analysis.

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Harwood, Steven J., and Hani Abdel-Nabi. "The Use of Monoclonal Antibodies for Radioscintigraphic Detection of Cancer." Journal of Pharmacy Practice 7, no. 3 (June 1994): 93–116. http://dx.doi.org/10.1177/089719009400700305.

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Beginning with Ehrlich's original “magic bullet” concept of 1904, the pioneering human trials in the late 1970s of Goldenberg and Mach using polyclonal antibodies, and the Nobel Prize-winning work of Kohler and Milstein in 1975 for developing monoclonal antibody (MAb) technology, there has been much interest in the use of antibodies for detecting and treating cancer. Although not the revolutionary breakthrough that was initially hoped for, marked progress has been made. The Food and Drug Administration (FDA) has recently approved the intact murine IgG, 111-indium CYT-103 (satumomoab, Oncoscint™ Cytogen, Princeton, NJ) for clinical use in detecting colorectal and ovarian cancer. However, the agent has been approved for only a single, one-time use, because patients developed an immune response (human anti-mouse antibody, or HAMA) that alters MAb biodistribution and may limit the clinical effectiveness of this agent when repeat studies are performed. Other MAbs reacting with a variety of antigens and targeting numerous tumors, including breast, lung, prostate, and melanomas, are currently undergoing large-scale clinical trials. To reduce induction of immune responses, many of the agents use immunoglobulin fragments [Fab, or F(ab)2] labeled with the short-lived isotope 99m-technetium used for most routine nuclear medicine diagnostic testing. Future developments will use even smaller fragments such as single chain antibodies or custom synthesized molecular recognition units (small peptides containing only the specific antigen combining site). Presented herein is an overview of the past results and an assessment of the current status of radioimmunoscintigraphy for various neoplasms.

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Al-Yousif, Yousif, Fahad Al-Majhdi, Cindy Chard-Bergstrom, Joe Anderson, and Sanjay Kapil. "Development, Characterization, and Diagnostic Applications of Monoclonal Antibodies against Bovine Rotavirus." Clinical Diagnostic Laboratory Immunology 7, no. 2 (March 1, 2000): 288–92. http://dx.doi.org/10.1128/cdli.7.2.288-292.2000.

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ABSTRACT Hybridomas secreting monoclonal antibodies (MAbs) against the Nebraska calf diarrhea strain of bovine rotavirus (BRV) were characterized. Indirect fluorescent-antibody assay, immunodot assay, and immunoprecipitation were used to select hybridomas that produced anti-BRV MAbs. Seven of the MAbs were shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot assay to be reactive with the BRV outer capsid protein, VP7, which has a molecular mass of 37.5 kDa. None of the seven MAbs were reactive with canine rotavirus, bovine coronavirus, or uninfected Madin-Darby bovine kidney cells. Two clones, 8B4 (immunoglobulin G2a [IgG2a]) and 2B11 (IgG1), were found suitable for use in an antigen capture enzyme-linked immunosorbent assay for detecting BRV in bovine fecal samples. Both were subtype A specific (G6 subtype) but did not react with all isolates of BRV group A.

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Skerritt, J. H., R. B. Gupta, J. L. Andrews, F. L. Stoddard, and N. K. Howes. "A rapid antibody-based test for Sec-2, a marker for the short arm of chromosome 2 of rye (2RS)." Genome 39, no. 5 (October 1, 1996): 1006–12. http://dx.doi.org/10.1139/g96-125.

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A simple monoclonal antibody-based screening test has been developed for the presence of translocations of the short arm of chromosome 2 of rye (2RS) with wheat chromosome 2B. 2RS encodes a set of about three polypeptides known as Mr 75 000 gamma-secalins. Use of the antibody test for these secalins enabled screening of several hundred seeds per day. The antibody could readily distinguish 2BL–2RS translocations and 2R substitutions from 1BL–1RS translocations or nontranslocation wheats. Use of the antibody in analysis of segregating progeny for Sec-2 in several wheat backgrounds was successful. Results with a selection of the seed population were checked using protein gel electrophoresis, with 100% correct confirmation. Key words : rye, wheat, seed proteins, translocation, diagnostic test.

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Vaidya, H. C., S. E. Porter, Y. Landt, D. P. Silva, D. N. Dietzler, and J. H. Ladenson. "Quantification of lactate dehydrogenase-1 in serum with use of an M-subunit-specific monoclonal antibody." Clinical Chemistry 34, no. 12 (December 1, 1988): 2410–14. http://dx.doi.org/10.1093/clinchem/34.12.2410.

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Abstract We have developed a rapid, one-step assay for measuring lactate dehydrogenase-1 (LD-1) activity in serum after extraction of LD-2, LD-3, LD-4, and LD-5 isoenzymes by an immobilized M-subunit-specific monoclonal antibody (D.8.1). In the assay, 100 microL of serum is mixed with 50 microL of a suspension of 0.8-micron-diameter latex particles coated with 30 micrograms of the monoclonal antibody D.8.1, then incubated at room temperature for 5 min. The latex particles, to which LD-2 through LD-5 are bound, are pelleted by centrifugation for 2 min at 12,000 X g, and the LD-1 activity is measured kinetically in the supernatant fluid. We optimized the assay for antibody immobilization, antibody concentration, and time and temperature of incubation. Serum bilirubin concentrations up to 0.33 g/L (0.56 mmol/L) did not interfere in the assay. Hemolysis interfered solely through LD-1 released from erythrocytes. The within-assay CV for low-concentration quality-control material (LD-1 33 U/L) was 3.5% (n = 9) and for high-concentration material (LD-1 185 U/L) was 1.9% (n = 8); the between-assay CVs for the two materials were 6.1% (n = 9) and 2.5% (n = 10), respectively. The LD-1 activity measured in 98 samples by our assay compared well with a two-step polyclonal antibody-based assay (Isomune-LD, Roche Diagnostic Systems; r = 0.998) and with an electrophoretic method (Paragon, Beckman Instruments; r = 0.956).

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Hou, Chong-lin, Yu Cao, Rong-hui Xie, Yi-zhen Wang, and Hua-hua Du. "Characterization and diagnostic use of a monoclonal antibody for VP28 envelope protein of white spot syndrome virus." Virologica Sinica 26, no. 4 (August 2011): 260–66. http://dx.doi.org/10.1007/s12250-011-3202-0.

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Tesfagaber, Weldu, Lulu Wang, Ghebremedhin Tsegay, Yibrah Tekle Hagoss, Zhenjiang Zhang, Jiwen Zhang, Haoyue Huangfu, et al. "Characterization of Anti-p54 Monoclonal Antibodies and Their Potential Use for African Swine Fever Virus Diagnosis." Pathogens 10, no. 2 (February 7, 2021): 178. http://dx.doi.org/10.3390/pathogens10020178.

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African swine fever (ASF) is a highly lethal hemorrhagic viral disease of domestic pigs caused by African swine fever virus (ASFV). Although a good advance has been made to understand the virus, a safe and effective vaccine against ASFV is still lacking and its eradication solely depends on its early and accurate diagnosis. Thus, improving the available diagnostic assays and adding some validated techniques are useful for a range of serological investigations. The aim of this study was to produce and characterize p54 monoclonal antibodies with an ultimate goal of developing a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for ASFV antibody detection. Five monoclonal antibodies against p54 protein expressed in Escherichia coli was generated and their characterizations were investigated. Furthermore, a competitive enzyme-linked immunosorbent assay (cELISA) based on a monoclonal antibody designated as 2A7 was developed. To evaluate the performance of the assay, a total of 365 pig serum samples (178 negative and 187 positive samples) were tested and a receiver-operating characteristic (ROC) analysis was applied to determine the cut-off value. Based on the ROC analysis, the area under the curve (AUC) was 0.982 (95% confidence interval: 96.9% to 99.4%), besides a sensitivity of 92.5% and a specificity of 98.9% was achieved when the percent inhibition of 20% was selected as a threshold. Moreover, the result showed an excellent agreement when compared to other commercially available blocking ELISA (kappa value = 0.912) and showed no reaction to other swine pathogens. Overall, the newly developed cELISA method offers a promising approach for a rapid and convenient ASFV serodiagnosis, which could be used as alternative to other serological assays for screening possible ASFV infection.

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Japolla, Greice, Ana Flávia Batista Penido, Greyciele Rodrigues Almeida, Luiz Artur Mendes Bataus, Jair Pereira Cunha Junior, Cristina Ribeiro, Sinji Borges Ferreira Tauhata, and Guilherme Rocha Lino de Souza. "Characterization of Monoclonal Antibodies against Bovine herpesvirus type 1 selected by Phage Display Technology." Semina: Ciências Agrárias 38, no. 6 (November 23, 2017): 3915. http://dx.doi.org/10.5433/1679-0359.2017v38n6p3915.

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The specificity of monoclonal antibodies (mAbs) to desired targets makes these molecules suitable for therapeutic and diagnostic uses against a wide range of pathogens. Phage display antibody libraries offer one method by which mAbs can be selected for, without the use of conventional hybridoma technology. In this work, phage display technology was used to construct, select and characterize a combinatorial single chain fragment variable (scFv) antibody library against bovine herpesvirus type 1 (BoHV-1) from the immune repertoire of chickens immunized with the virus. In silico analysis of the hypervariable domains of the antibody heavy chains revealed a high frequency of scFv fragments with low variability, suggesting that selection had probably been carried out and favored by a few im-munogenic viral antigens. The reactivity of the scFv fragments selected against BoHV-1 was demon-strated by Phage-ELISA. A significant increase in antibody reactivity to the target was observed after six rounds of library selection, showing its potential use as a molecule for BoHV-1 diagnosis. The strategy described here opens up a field for the use of phage display as a tool for selection of mono-clonal antibodies that could be used for theranostic applications against infectious and parasitic dis-eases of veterinary interest.

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Journal articles on the topic 'Monoclonal antibody probes Diagnostic use'

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