Дисертації з теми "Phage library"
Оформте джерело за APA, MLA, Chicago, Harvard та іншими стилями
Ознайомтеся з топ-50 дисертацій для дослідження на тему "Phage library".
Біля кожної праці в переліку літератури доступна кнопка «Додати до бібліографії». Скористайтеся нею – і ми автоматично оформимо бібліографічне посилання на обрану працю в потрібному вам стилі цитування: APA, MLA, «Гарвард», «Чикаго», «Ванкувер» тощо.
Також ви можете завантажити повний текст наукової публікації у форматі «.pdf» та прочитати онлайн анотацію до роботи, якщо відповідні параметри наявні в метаданих.
Переглядайте дисертації для різних дисциплін та оформлюйте правильно вашу бібліографію.
Bell, Matthew John. "Phage display of a cDNA library from Arabidopsis thaliana." Thesis, Coventry University, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.393652.
Повний текст джерелаKeklikian, Artine. "Construction of a Synthetic Human VL Phage Display Library and Isolation of Potential Neuropilin-1-specific VL Therapeutics from the Library." Thèse, Université d'Ottawa / University of Ottawa, 2011. http://hdl.handle.net/10393/20197.
Повний текст джерелаClayton, Ruth. "Generation and characterisation of human sperm antibodies by combinatorial phage display library technology." Thesis, University of Sheffield, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.286988.
Повний текст джерелаHald, Rikke. "Generation and characterisation of a naive human antibody phage display library : a resource for clinically relevant reagents /." Cph. : Department of Pharmacology, The Danish University of Pharmaceutical Sciences, 2004. http://www.dfh.dk/phd/defences/rikkehald.htm.
Повний текст джерелаPrendergast, D. "Discovery of tumour necrosis factor receptor-1 (p55) binding peptides using a phage display library." Thesis, Queen's University Belfast, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.368468.
Повний текст джерелаSaxena, Abhishek. "Construction of immune scFv M13 phage display library and isolation of anti-glycan monoclonal antibodies." Thesis, University of Aberdeen, 2013. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=203295.
Повний текст джерелаLim, Theam Soon [Verfasser]. "Parameters affecting phage display library design for improved generation of human antibodies / Theam Soon Lim." Berlin : Freie Universität Berlin, 2009. http://d-nb.info/1023748355/34.
Повний текст джерелаCortini, Andrea. "Serum profiling and autoantibodies identification in Multiple Sclerosis using epitope and CSF IgG phage display libraries." Doctoral thesis, Università degli studi di Trieste, 2009. http://hdl.handle.net/10077/3073.
Повний текст джерелаMultiple sclerosis (MS) is considered the prototype of inflammatory autoimmune diseases of the central nervous system (CNS). The typical feature of the disease is the plaques of demyelination. The evolution of the plaque lesion in MS implicates an inflammatory phase followed by a recovery of functional myelin; a second step is the chronic progressive disease with axonal loss. The earlier phase of MS may be mediated by an autoimmune reaction. Whereas the role of T cells in MS pathogenesis is well established, the role of B cells and autoantibodies in demyelination and plaque formation is still unresolved. However several evidences suggest a contribute of autoantibodies in MS pathogenesis. B cells and myelin specific autoantibodies are present in the sclerosis plaques, and there is an increased production of immunoglobulin (Ig) in the cerebrospinal fluid (CSF) of more than 90% of MS patients . Typically these Ig present an oligoclonal pattern and sequencing of oligoclonal IgG showed extensive somatic mutations suggesting B cell clonal expansion and a specific antigen-driven immune response. The most extensively studied putative autoantigens are components of CNS myelin (myelin basic protein MBP, proteolipid protein PLP, myelin oligodendrocyte glycoprotein MOG). The autoantibodies in MS recognize both linear and conformational epitopes, but at present the conformational epitopes of myelin proteins have not been identified. For example, in MS, the T-cell receptors of autoreactive T lymphocytes recognize various peptides of the MBP, and, in EAE, the anti-MOG antibodies recognize only conformational epitopes. Furthermore, the progression of MS is accompanied by the decline of primary T-cell autoreactivity and by the concurrent emergence of neo-autoreactivity (epitope spreading). However recent investigation have showed that no myelin antigens, like neuron-specific enolase (NSE), retinal arrestin, beta-arrestin, may also have a role in MS pathogenesis. Autoimmunity against these antigens may be linked to neurodegeneration, defective remyelination, and predisposition to uveitis in multiple sclerosis. Several strategies, involving the phage display technology, have been employed in the attempt to discover the antigen that drives the immune response in MS. A first strategy depends on the cloning of IgG repertoire of MS patients in a phage display library screened with brain sections or known antigens. Another strategy involves large phage display libraries of random peptides screened with IgG of CSF in order to identify peptides recognized by antibodies present in CSF of MS patients. Phage display is a technique which involves the coupling of phenotype to genotype in a selectable format. It has been extensively used in molecular biology to study protein-protein interactions and to select antibodies against a wide range of different antigens. In this project we have proposed: 1. to study the autoimmune response in MS by using the phage display for the expression of antibodies involved in the disease. We wanted to make a ScFv library from B cells of CSF of different multiple sclerosis patients, to employ as tool to select a phage display Human Brain cDNA library for the identification of new antigens recognized by the immune sistem in MS patients. 2. To produce single gene mini library of putative antigens (MBP,PLP, MOG) for the generation of epitope chips to use for serotyping the immune response in different patients 3. To investigate the feasibility to use a single gene phage display mini-library as tool for epitope mapping (both linear and conformational) of novels autoantigens 4. To investigate the role of NSE(neuron specific enolase), a new possible no myelin autoantigen in multiple sclerosis, in the pathogenesis of the disease and the usefulness as possible diagnostic marker. Results: Scope 1 B-cells from liquor of two MS patients were centrifuged and the total RNA was extracted from the pellets. Total RNA was retrotranscripted and variable region of heavy and light chain of the antibodies were amplified by PCR. Heavy chain and light chain were assorted and assembled before to be cloned in the phagemid vector pDAN 5. A 2x104 independent clones library was obtained and analyzed by PCR and fingerprinting. A diversity of 30,8% for heavy chain and 72,7% for light chain was established. ScFv library was used to select a phage display Human Brain cDNA library. 17 clones with an high reactivity were obtained and after sequencing 6 clones on 17 have shown to be the same antigen(antigen A ); the reactivity on other two antigens obtained with the selection (antigen B and C) of CSF from 18 MS patients and 16 patients with other neurological disease (OND) was tested by ELISA to evaluate diagnostic value of this protein. The results shown that SM response was statistically different from OND response; the ELISA test gave a specificity of 94,12% and a significance of 53,85 %. The reactivity for the antigen B was also evaluated on sera of MS patients and controls. The MS response was statistically different from OND response and shown a specificity of 97,44% and a significance of 58,62 %. Scope 2&3 We have generated three single gene mini libraries of the major antigens in MS (MBP, MOG and PLP); cDNA of each gene was obtained by RT-PCR and after fragmentation cloned in a phagemid vector (pEP1) to obtain a mini-library for each gene. We have obtained a 2x105 for MBP, 2.4x104 for MOG and 1.6x106 for PLP independent clones library. MBP and MOG libraries were characterized by PCR and fingerprinting. Sequencing analysis shown that the entire MBP transcript variant 7 mRNA (664-1177 nt) and MOG isoform alpha 1 mRNA (262-918 nt) were represented in the respective library. To testing the capacity of selecting a single epitope from our libraries, we have performed a selection test with a commercial monoclonal antibody that recognize MBP 82-98 epitope; after three selection panning all selected clones contain the nucleotidic sequence 906- 956 nt (MBP transcript variant 7 mRNA) which encodes the immunogenic epitope recognized by the monoclonal antibody. Scope 4 The reactivity of sera from 31 MS patients and 14 healthy controls was tested by ELISA on NSE ; statistical analysis of the results shown that the two populations were significantly different.
XXI Ciclo
1981
Bosompem, Amma N. "Isolation of an anti-CD20 single chain variable fragment from a naïve human phage-scFv library." Connect to this title online, 2007. http://etd.lib.clemson.edu/documents/1202410076/.
Повний текст джерелаMarson, Lorena. "Phage-display epitope library development for biomarkers identification in autoimmune diseases of the Central Nervous System." Doctoral thesis, Università degli studi di Trieste, 2012. http://hdl.handle.net/10077/7405.
Повний текст джерелаThe principal aim of my PhD was the setting of a protocol for the creation of phage libraries to display cDNA fragments encoding real ORF sequences, that could correspond to potential epitopes. A similar phage display library contains all the potential ORF repertoire of a cell or tissue. This tool can be specially used in the study of autoimmune diseases to perform different kind of analysis, such as the identification of epitopes involved in pathological reaction, the comparison between healthy and pathological conditions, or between different pathological conditions. A complex protocol was developed. It provides for: cDNA normalization, cDNA fragmentation to obtain peptides with useful size, and ORF enrichment to obtain really coding fragments. With this system we have created a epitopes library from Human brain mRNA.
Il principale obiettivo del mio lavoro di ricerca è la messa a punto di un protocollo per la costruzione di librerie fagiche di frammenti di cDNA codificanti per frammenti ORF, e che quindi potrebbero corrispondere a potenziali epitopi. Questo tipo di librerie contengono, potenzialmente, tutto il repertorio ORF di una cellula o di un tessuto e possono quindi essere utilizzate nello studio di malattie autoimmuni al fine di identificare nuovi epitopi coinvolti nella risposta immunitaria, di fare un confronto tra lo stato patologico e quello sano o tra diverse condizioni patologiche. Abbiamo quindi messo a punto un complesso protocollo che prevede: la normalizzazione del cDNA, la sua frammentazione per ottenere peptidi di dimensioni opportune, e l'arricchimento in frammenti realmente codificanti. Con questo sistema abbiamo realizzato una libreria di epitopi a partire da mRNA di cervello umano.
XXIV Ciclo
1984
Abi, Ghanem Daad Ali. "Phage display selection of recombinant antibodies derived from a chicken immune library against cryopreserved Eimeria tenella sporozoites." [College Station, Tex. : Texas A&M University, 2006. http://hdl.handle.net/1969.1/ETD-TAMU-1847.
Повний текст джерелаSorsa-Leslie, Tarja. "Combining phage display antibody library and bioassay technologies to identify candidate gonadotropin surge attenuating factor (GnSAF) molecules." Thesis, University of Aberdeen, 2005. http://digitool.abdn.ac.uk/R?func=search-advanced-go&find_code1=WSN&request1=AAIU212837.
Повний текст джерелаNguyen, Hai Phu. "Combination of hu-PBL-SCID mice and scFv phage display library, an effective alternative for hu-mAb production." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2001. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/NQ58617.pdf.
Повний текст джерелаFehrsen, Jeanni. "Isolation of antigenic peptides of Cowdria ruminantium and their encoding genes using a genome-derived phage display library." Thesis, Rhodes University, 2003. http://hdl.handle.net/10962/d1003979.
Повний текст джерелаBessa, Juliana Mattos de Almeida. ""Determinação de alvos antigênicos na doença reumática cardíaca utilizando phage display"." Universidade de São Paulo, 2006. http://www.teses.usp.br/teses/disponiveis/5/5146/tde-12042006-164408/.
Повний текст джерелаRheumatic heart disease (RHD) patients develop valvar lesions with CD4+ T lymphocytes infiltrating the heart. Molecular mimicry between streptococcal and cardiac proteins recognized by these T cells may explain these auto-aggressive lesions. In the present work we used a Phage Display peptide library to identify cardiac antigens which could be recognized by two heart infiltrate T cell lines and by a T cell clone derived from one of the lines which were isolated from RHD patients. Using the protein data bank to analyse the phage peptides, we observed that many sequences showed homology with cardiac myosin, laminin, vimentin and other coiled-coil proteins, suggesting the involvement of these proteins in the autoimmune process of RHD. Other inflammatory molecules such as cytokines and integrins were also identified
Igloi, Zsofia. "Identifying novel interaction partners for the hepatitis C virus NS5A protein by screening a human SH3 domain phage display library." Thesis, University of Leeds, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.574514.
Повний текст джерелаCaucheteur, Déborah. "Nouveau format de banques d’anticorps recombinants humains pour un criblage fonctionnel à grande échelle." Thesis, Montpellier, 2018. http://www.theses.fr/2018MONTT012.
Повний текст джерелаSince 2000, monoclonal antibodies (mAb) have become essential and routine drugs in therapy and particularly in oncology. The field continues to grow very quickly and given the abundance of molecules available, it is increasingly important to bring innovative molecules with a high added value for therapy.Two main approaches are used to select these mAbs: hybridoma technology using normal or humanized mice; display systems such as phage-display. The major interests of phage display are the speed of mAb development, the facilities offered by E. coli and the easy access to protein engineering techniques. Typically, antibodies are first selected on their ability to bind to the antigen, and then tested for their functional efficiency in cellular models. However, only a part of the activity of antibodies is explained by their binding to the antigen, and the therapeutic activity also depends strongly on their ability to recruit the immune system (ADCC) and activate the complement cascade (CDC).This thesis project consists in the development of a new recombinant antibody library format combining the power of phage display selection with functional screening in a whole IgG format produced in eukaryotic cells. This new system is based on hybrid promoter and signal peptide regions allowing expression both in prokaryotic and eukaryotic cells, and a site-specific recombination event that exchanges the Fab between the display vector and the chromosome of an especially developed mammalian cell line resulting in the secretion of a monoclonal human antibody by the cell. The usual approach of recloning one by one from E.Coli vector to an IgG format is no more needed since it is done directly by transfection. This new system makes possible to couple selection by phage display with a direct functional screening of a large population of human monoclonal clones
Ndlovu, Siphumelele. "The isolation of single chain variable region fragments (scFvs) from a phage display library, and expression of the isolated scFvs in Nicotiana benthamiana." Master's thesis, Faculty of Science, 2020. http://hdl.handle.net/11427/32303.
Повний текст джерелаBandmann, Nina. "Rational and combinatorial genetic engineering approaches for improved recombinant protein production and purification." Doctoral thesis, Stockholm : Bioteknologi, Kungliga Tekniska högskolan, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4318.
Повний текст джерелаHubert, Ágnes [Verfasser], Wolfgang [Akademischer Betreuer] Baumeister, and Sevil [Akademischer Betreuer] Weinkauf. "Needle in the haystack: Protein complex purification from Thermoplasma acidophilum with a phage display library / Ágnes Hubert. Gutachter: Sevil Weinkauf ; Wolfgang Baumeister. Betreuer: Wolfgang Baumeister." München : Universitätsbibliothek der TU München, 2013. http://d-nb.info/103576671X/34.
Повний текст джерелаMundin, Georgia Sabio Porto. "Identificação de marcadores moleculares para células T reguladoras humanas com perfil CD4+CD25+ por phage display." Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/5/5146/tde-28012009-131608/.
Повний текст джерелаThere are consistent data in literature indicating that thymic CD4+CD25+ cells play an important role in immune regulation and are continuously developed as an independent lineage in the thymus. These cells are known as natural regulatory T cells. Several questions about these cells remain unanswered, such as how they are generated, what is determinant in their regulatory function and which specific molecular markers can be used to identify them. Taking this into consideration, our aim was to identify new potentially important molecules in the development and/or supressive function of natural regulatory T cells, both in the thymus and in CD4+CD25+ thymocytes. For this, the phage display technique was employed, with a peptide phage library and thymic specimens obtained from children who underwent corrective cardiac surgery at the Heart Institute (InCor), in São Paulo. The search for these molecules was separately performed in 3 types of biological material: thymic tissue, thymocytes and CD4+CD25+ thymic cells. In the first stage, the phage peptide-library was incubated with a pool of PBMC (peripheral blood mononuclear cells). After the incubation, phages bound to PBMC were discarded (pre-clearing). In the second stage, unbound phages were incubated with either total thymocytes or CD4+CD25+ thymic cells. The pre-clearing stage was not perfomed in the thymic tissue. The phages obtained with after incubation with the different biological materials were recovered in E. coli culture and used in additional cycles of selection. After three rounds of selection, the recovered phages from the total thymocytes, from thymic tissue and thymocytes CD4+CD25+ were sequenced and their ligands identified. Among the phages selected for validation one ligand of thymic tissue: M2C and one ligand of CD4+CD25+ thymocytes: R2A present similarity to two proteins associated to the metabolism of Vitamin D3, a molecule involved in imunoregulation and toelrance induction in several experimental models. However, there are no data in the literature concerning the possible role of this moelcule in natural regulatory T cells. In the molecular validation of theses phages, although some variability between the diffeterent assays we have verified by ELISA, that the phages present preferential binding to the 1,25 dhydroxyvitamin D3, the active form of Vitamin D3. However, in the functional validation assays, the influence of the Vitamin D3 in the differentiation of these cells could not be consistently confirmed since we could observe an increase in the number of CD4+CD25+ cells cultured with vitamin D in only a few experiments. The ligand-receptor molecules we have defined in this study may have relevant implications in the development of CD4+CD25+ regulatory T cells in the thymus
Sulaiman, Lanre Precieux Kabir. "Seleção de motivos semelhantes a Papilomavírus, a partir de bibliotecas de phage display, que apresentem potencial aplicação translacional." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/5/5155/tde-29012018-133017/.
Повний текст джерелаThe link between high-risk human papillomavirus and cervical cancer is well established. Despite the existence of prophylactic vaccines against infections by the most common types of HPV, therapeutic alternatives are limited for infections and tumors caused by these viruses. We found some homology motifs for high-risk HPV proteins during the immune-panning of a phage display library with sera from HPV-16- seropositive participants of the Ludwig-McGill cohort. After enrichment of the sequences, the recombinant bacteriophages were purified and amplified for use as immunogens. Using a prophylactic approach, we vaccinated experimentally immunocompetent mice with one of our recombinant bacteriophages using the insertless bacteriophage as a control. These mice were then challenged with TC-1 tumor cells (HPV-16 positive), and the immune responses triggered during tumor progression were evaluated. We also used a therapeutic approach where mice were first injected with tumor cells and immunized with the bacteriophage after tumor establishment. Tumor growth was monitored and tumors, spleen and lymph nodes were evaluated for the quantity and quality of the immune response. ELISA tests revealed that all vaccinated mice responded to immunization with the different bacteriophages. Tumor growth was significantly reduced in prophylactic and therapeutic immunizations, although tumor reduction was minimal when mice were treated 9 days after TC-1 cells grafting. The reduction in tumor growth also translated into a significantly greater survival for the immunized mice. Cell infiltration studies did not reveal changes in several immune subpopulations, but an upward trend in cytotoxic T lymphocytes was observed in mice immunized with PEP1 (insert-containing bacteriophage). The importance of this increase in CD8 in the observed reduction of tumor growth was confirmed using CD8-knockout mice, where the previously observed reduction of tumor growth was abolished. An increase in CD8:CD4 rate was observed in the immunized mice and this is an indication of a cytotoxic tumor environment. Cell proliferation assays to test the antigen specificity of lymphocytes from immunized mice were, however, inconclusive; likewise, we could not change the pattern observed with the use of CpG adjuvant. The usefulness of the phage display technique was also observed in this experimental work. Additional studies to understand the mechanism of action of these recombinant phages in the control of HPV tumor growth and its immunostimulatory potential are warranted
Zou, Jun. "Characterization of peptides and phage that bind galectin-3 selected from bacteriophage display libraries a study of the role of galectin-3 in metastasis-associated cancer cell adhesion /." Diss., Columbia, Mo. : University of Missouri-Columbia, 2005. http://hdl.handle.net/10355/4149.
Повний текст джерела"December 2005" The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. Includes bibliographical references.
Bernedo-Navarro, Robert Alvin 1975. "Obtenção de peptídeos com capacidade inibitória da ação citotoxigênica das toxinas Stx de Escherichia colia partir de bibliotecas de phage display." [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/314705.
Повний текст джерелаTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
Made available in DSpace on 2018-08-23T11:52:53Z (GMT). No. of bitstreams: 1 Bernedo-Navarro_RobertAlvin_D.pdf: 2646572 bytes, checksum: 02b50bc28d5be6cddb78abfe621a047c (MD5) Previous issue date: 2013
Resumo: Escherichia coli produtora de toxina de Shiga (STEC) é um grupo de importantes patógenos para humanos. Essas bactérias são relacionadas a várias doenças, como por exemplo, Síndrome Urêmica Hemolítica e produzem potentes toxinas denominadas toxinas de Shiga. Essas toxinas, tanto Stx1 quanto Stx2, compartilham um receptor celular comum, a globotriaosilceramida (Gb3) e exibem a mesma atividade biológica intracelular. O desenvolvimento de novos agentes neutralizantes dos danos induzidos por Stx pode representar uma estratégia promissora para o tratamento das doenças causadas por STEC em humanos. No presente estudo, nós desenvolvemos peptídeos sintéticos que exibem atividade neutralizante contra a citotoxicidade induzida por Stx tanto in vitro quanto in vivo e, além disso, que se ligam eficientemente ao receptor Gb3. O peptídeo P12-26 compete eficientemente com Stx2 para a ligação ao Gb3 in vitro. Além disso, os peptídeos PC7-12, P12-26 e PC7-30 inibiram a citotoxicidade de Stx1 e Stx2 em células Vero. Nós observamos que o peptídeo PC7-30 em forma de loop e o peptídeo P12-26 que é linear produziram as maiores porcentagens de inibição de Stx1 e Stx2 em células Vero, respectivamente. No entanto, o peptídeo P12-26 não inibiu a letalidade em camundongos, enquanto que o peptídeo PC7-30 inibiu a letalidade causada pela toxina Stx1. Nossos resultados indicam que os peptídeos P12-26 e PC7-30 são candidatos promissores para o desenvolvimento de agentes terapêuticos contra as doenças em seres humanos causadas por STEC
Abstract: Shiga toxin-producing Escherichia coli strains are important pathogens for humans. These bacteria are linked with severe diseases such as hemolytic uremic syndrome and produce potent known as Shiga toxins. These toxins, Stx1 and Stx2, share a common cellular receptor called globotriaosylceramide (Gb3) and exhibit the same intracellular biological activity. The development of new neutralizing agents for Stx-induced damage may represent a promising strategy for the treatment of diseases caused by STEC infections. In this study, we developed synthetic peptides that exhibit neutralizing activity against Stxinduced cytotoxicity both in vitro and in vivo and that bind efficiently to the Gb3 receptor. The peptide P12-26 competed efficiently with Stx2 for binding to Gb3 in vitro. Moreover, the peptides PC7-12, P12-26 and PC7-30 inhibited the cytotoxicity of Stx1 and Stx2 in Vero cells. We observed that the loop-constrained peptide PC7-30 and linear peptide P12-26 produced higher percentages of inhibition of Stx1 and Stx2 in Vero cells, respectively. However, the peptide P12-26 did not inhibit lethality in mice, whereas the loopconstrained peptide PC7-30 inhibited the lethality caused by Stx1. Our results indicate that the peptides P12-26 and PC7-30 are promising candidates for the development of therapeutic agents against diseases caused by STEC in humans
Doutorado
Bioquimica
Doutor em Biologia Funcional e Molecular
Lehtiö, Janne. "Functional studies and engineering of family 1 carbohydrate-binding modules." Doctoral thesis, KTH, Biotechnology, 2001. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-3211.
Повний текст джерелаThe family 1 cellulose-binding modules (CBM1) form a groupof small, stable carbohydrate-binding proteins. These modulesare essential for fungal cellulosedegradation. This thesisdescribes both functional studies of the CBM1s as well asprotein engineering of the modules for several objectives.
The characteristics and specificity of CBM1s from theTrichoderma reeseiCel7A and Cel6A, along with severalother wild type and mutated CBMs, were studied using bindingexperiments and transmission electron microscopy (TEM). Datafrom the binding studies confirmed that the presence of onetryptophan residue on the CBM1 binding face enhances itsbinding to crystalline cellulose. The twoT. reeseiCBM1s as well as the CBM3 from theClostridium thermocellumCipA were investigated by TEMexperiments. All three CBMs were found to bind in lineararrangements along the sides of the fibrils. Further analysesof the bound CBMs indicated that the CBMs bind to the exposedhydrophobic surfaces, the so called (200) crystalline face ofValoniacellulose crystals.
The function and specificity of CBM1s as a part of an intactenzyme were studied by replacing the CBM from the exo-actingCel7A by the CBM1 from the endoglucanase Cel7B. Apart fromslightly improved affinity of the hybrid enzyme, the moduleexchange did not significantly influence the function of theCel7A. This indicates that the two CBM1s are analogous in theirbinding properties and function during cellulosehydrolysis.
The CBM1 was also used for immobilization studies. Toimprove heterologous expression of a CBM1-lipase fusionprotein, a linker stability study was carried out inPichia pastoris. A proline/threonine rich linker peptidewas found to be stable for protein production in this host. Forwhole bacterial cell immobilization, theT. reeseiCel6A CBM1 was expressed on the surface of thegram-positive bacteria,Staphylococcus carnosus. The engineeredS. carnosuscells were shown to bind cellulosefibers.
To exploit the stable CBM1 fold as a starting point forgenerating novel binders, a phage display library wasconstructed. Binding proteins against an amylase as well asagainst a metal ion were selected from the library. Theamylase-binding proteins were found to bind and inhibit thetarget enzyme. The metal binding proteins selected from thelibrary were cloned on the surface of theS. carnosusand clearly enhanced the metal bindingability of the engineered bacteria.
Keywords: cellulose-binding, family 1carbohydrate-binding module, phage display, bacterial surfacedisplay, combinatorial protein library, metal binding, proteinengineering,Trichoderma reesei, Staphyloccus carnosus.
Grimm, Sebastian. "Ribosome display for selection and evolution of affibody molecules." Doctoral thesis, KTH, Molekylär Bioteknologi, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-33191.
Повний текст джерелаQC 20110506
Shahsavarian, Melody. "Genesis of immune diversity and selection of catalytic antibodies : a new investigation." Thesis, Compiègne, 2015. http://www.theses.fr/2015COMP2215/document.
Повний текст джерелаCatalytic antibodies (or abzymes) have been the focus numerous studies for some decades and have been produced with the ability to catalyze a wide range of reactions. They have also been discovered naturally in normal physiological and pathological conditions, notably on the background of autoimmune disease. Some have beneficial effects and others are detrimental to individual’s health. Hence, the origin of abzymes and their role in the immune response are ambiguous and must be enhanced. We have developed a novel strategy for the study of abzymes based on the phage display technology. We have constructed 4 libraries representing 4 murine immune repertoires with different genetic backgrounds and immunological states : healthy and naïve, healthy and immunized, autoimmune and naïve , and autoimmune and immunized. The strategies for the amplification and cloning of the immunoglobulin (lg) variable regions have been designed to optimize the size and diversity of the libraries. We have been able to pool the four libraries to create a large repertoire of size 2.7x109. After sequence analysis, we have found a number of statistically significant differences between the libraries. We have then used two strategically chosen targets to select for antibodies endowed with β lactamase activity : a cycle peptide and a penam sulfone, both inhibitors of the enzyme. We have selected for a total of 5 lgs endowed with β lactamase activity. The selected abzymes have different amino acid sequences. 3D modeling has provides insights on potential active sites demonstrating the ability of different structures to maintain the β lactamase activity and confirming the flexibility of the active site
Harris, Jasmine K. "Bioconjugation of aminated DNA as a method of rapid polymer library generation for Corona Phase Molecular Recognition." Thesis, Massachusetts Institute of Technology, 2018. http://hdl.handle.net/1721.1/119063.
Повний текст джерелаCataloged from PDF version of thesis.
Includes bibliographical references (page 39).
An experimental study was performed to determine the effects on Corona Phase Molecular Recognition (CoMoRe) of bioconjugating a host of small molecules to DNA wrapped single-walled carbon nanotubes. In addition, the study observed the effects of the DNA sequence length on the subsequent effectiveness of the small molecules to alter the corona phase. The conjugation of small molecules was shown to alter both the intensity and the position of the fluorescence and absorbance profile. The length of the DNA sequence was found to change the small molecule's ability to alter the fluorecence spectra of the wrapped nanotubes. The EDC/sulfo-NHS reaction was done to conjugate the small molecules to two identical DNA sequences with varying lengthes. Through the methods of ultraviolet-visibile-near infrared absorption spectroscopy, near infrared fluorescence spectroscopy, and high-performance liquid chromatography characterization and structural analysis were performed. The results showed the successful conjugation of the small molecules to the amino-modified DNA and an alteration in the corona phase. The small molecules were found to bind to the DNA at multiple locations and the length of the sequence was found to have an effect on the corona phase.
by Jasmine K. Harris.
S.B.
Hwang, Sung Hee. "I. Combinatorial solid-phase synthesis of isoxasolinopyrroles. II. OBOC small molecule combinatorial library encoded by halogenated mass-tags /." For electronic version search Digital dissertations database. Restricted to UC campuses. Access is free to UC campus dissertations, 2004. http://uclibs.org/PID/11984.
Повний текст джерелаYamakawa, Tatsuya. "Screening of human cDNA library reveals two differentiation-related genes, HHEX and HLX, as promoters of early phase reprogramming toward pluripotency." Kyoto University, 2016. http://hdl.handle.net/2433/217142.
Повний текст джерелаRuda, Marcus. "Design and synthesis of steroid mimetic libraries using solid phase techniques /." Stockholm, 2004. http://diss.kib.ki.se/2004/91-7140-049-4/.
Повний текст джерелаHuang, Adela Ya-Ting. "Advancing dendrimer synthesis : solid-phase and self-assembly approach." Thesis, Aix-Marseille, 2017. http://www.theses.fr/2017AIXM0146.
Повний текст джерелаDendrimers hold great promise for wide applications thanks to their unique structural architecture and multivalent cooperativity. However, dendrimer synthesis often suffers from structural defects caused by incomplete reactions and difficulties associated with purification. Consequently, alternative synthetic approaches to overcome the limitations of current dendrimer synthesis are in high demand.My first PhD project mainly focuses on establishing novel strategies and methodologies for solid-phase dendrimer synthesis with advantages of convenient complete synthesis and easy purification procedures. We first developed a new and concise solid-phase synthesis of PAMAM dendrimers based on the adoption of peptide synthesis chemistry. We then constructed a small library of triazine dendrimers varying in generations and surface groups with a view to rapidly synthesizing dendrimers with structural diversity. We also strived to synthesize poly(aminoester) dendrimers although we had difficult to get it thorough.My second PhD program aims to apply the self-assembly approach for constructing supramolecular dendrimer theranostics. A small DOTA-conjugated amphiphilic dendrimer with Gd(III)-chelation was synthesized and self-assembled into supramolecular nanomicelles to encapsulate the anticancer drug doxorubicin. The obtained system constitutes a multivalent nanotheranostic to combine imaging purpose with therapeutic utility.In summary, my PhD program mainly contributes to elaborating strategies for dendrimer synthesis using both solid-phase method and self-assembly approach in the view to realizing and broadening their applications in the arenas of biomedical and material sciences
Chen, Xingguo Ronald. "Pin1 Catalytic and WW Domain Ligands." Diss., Virginia Tech, 2011. http://hdl.handle.net/10919/37824.
Повний текст джерелаPh. D.
Nilsson, Jonas. "Design, Synthesis and Characterization of Small Molecule Inhibitors and Small Molecule : Peptide Conjugates as Protein Actors." Doctoral thesis, Linköpings universitet, Organisk Kemi, 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-3943.
Повний текст джерелаSubramaniam, Raja. "Simplified Routines for Sample Preparation and Analysis of Chemical Warfare Agent Degradation Products." Doctoral thesis, Umeå universitet, Kemiska institutionen, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-54639.
Повний текст джерелаAftab, Obaid. "Towards High-Throughput Phenotypic and Systemic Profiling of in vitro Growing Cell Populations using Label-Free Microscopy and Spectroscopy : Applications in Cancer Pharmacology." Doctoral thesis, Uppsala universitet, Cancerfarmakologi och beräkningsmedicin, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-234565.
Повний текст джерелаLeslie, Susan Elder. "An examination of the information behaviour of new entrepreneurs in the start-up phase of a business submitted to the School of Information Management, Victoria University of Wellington in partial fulfilment of the requirements for the degree of Master of Library and Information Studies /." ResearchArchive@Victoria e-Thesis, 2009. http://hdl.handle.net/10063/1271.
Повний текст джерелаJoo, Sang Hoon. "Synthesis and screening of support-bound combinatorial cyclic peptide and free C-terminal peptide libraries." Columbus, Ohio : Ohio State University, 2007. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1195561420.
Повний текст джерелаLIN, MING-CHUN, and 林明君. "Ractopamine Detection Through Phage Display Peptide Library." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/48211792124612562148.
Повний текст джерелаChou, Ying-Hsiu, and 周盈秀. "Construction of human phage-displayed scFv library." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/13301253686668832243.
Повний текст джерела國立成功大學
醫學檢驗生物技術學系碩博士班
97
Antibodies are applied not only for diagnosis, but also in research and clinical treatments. In order to generate high-affinity monoclonal antibodies, three techniques have been developed including hybridoma engineering, transgenic mice and phage-displayed antibody libraries. Although the hybridoma technology is the major tool for monoclonal antibody production, it still has some problems. For example, the methodology cannot be curable directly in human disease. Hence, the phage antibody display system has become more attractive to scientists. We attempted to develop a na��ve human single chain antibody (scFv) library by phage display system. We first extracted total RNA from the leukocytes of peripheral blood from 290 volunteers. The heavy chain and light chain cDNA fragments were synthesized by reverse transcription reaction. Thirty-one forward and reverse primers were used for the cloning of heavy chain variable regions (VH) and light chain variable regions (VL) fragments from the cDNA fragments. The cloned heavy and light chain variable regions were amplified and linked with linkers by overlapping PCR. The linkers encoded the 15 amino acids (Gly4Ser)3 that have overhangs of perfect complementarity with VH and VL genes. The purified scFv fragments were ligated to the phagemid pIGT5 for electroporation into Escherichia coli HB2151 competent cells. In order to increase the binding avidity of scFv during bio-paining, we had constructed pIGT5 phagemid which expressed trypsin and enterokinase cutting site between pIII and scFv. We also established Ex-phage in which the pIII protein could not be synthesized in non-amber suppressed strain owing to the amber codon before pIII. Hence, the binding phage can be released after enzymatic digestion and amplified for next paining. After the library constructed, we will apply this antibody library to discover new carbohydrate antibodies.
Lin, Shu-Jian, and 林淑娟. "Construction of scFv Phage Display Library Against Diphenylamine." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/92251249392714967412.
Повний текст джерела國立中興大學
生命科學院碩士在職專班
92
Diphenylamine is an antioxidant, having the skeleton structure of non-steroidal anti-inflammatory drugs. In the use of agriculture, it is as a dipping solution to prevent superficial scald in the surface of apple. Diphenylamine also is a fungicide and was used in the pesticides with high concentration. It is an important derivative of BXT, and often used in hair color dye. It is toxic, possible mutagen and teratogen, and harmful in contact with skin. Because diphenylamine is a hapten, in this experiment, we use conjugate reagent to synthesize the bovine serum albumin with diphenylamine together as an immunogen. After immunization, and confirmed with ELISA, we take out the spleen of mouse, and use a series of degenerate primers covering recently known murine immunoglobulin gene family and performing RT-PCR to amplify variable region of heavy chain (VH) and light chain (VL) gene. The amplified VH and VL DNA fragments were assembled to perform single chain fragment of variable region (scFv). The scFv DNA were cloned to the pHEN2 vector, and then transformed to E. coli. We use the technique of phage display system to construct the library of scFv for further selection. The constructed antibody phage display library was confirmed the diversity by BstNI restriction fingerprinting analysis.
Yi-WenChang and 張憶文. "Discovery of antibodies by phage display human scFv antibody library." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/15750785477965469165.
Повний текст джерелаHu, Ting-Yuan. "Titanium-binding Peptide Isolated from Phage-displayed Random Peptide Library." 2007. http://www.cetd.com.tw/ec/thesisdetail.aspx?etdun=U0001-3007200716354000.
Повний текст джерелаHu, Ting-Yuan, and 胡庭元. "Titanium-binding Peptide Isolated from Phage-displayed Random Peptide Library." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/93998814871702072948.
Повний текст джерела臺灣大學
臨床牙醫學研究所
95
Since the introduction of titanium root-form dental implant in 60’s, it has become a widely used treatment modality in rehabilitation of completely or partially edentulous patients. However, clinicians still face the failure of implant treatment or its complication, such as progressive marginal bone loss, recession of marginal mucosa, or absence of osseointegration, inspite of its reported high success and survival rates in the literatures. In late 90’s, the application of biomimetic modification has been introduced to overcome the problems because of the advances in the understanding of the biology of bone healing process at molecular level. However, there is only marginal success in this field of research after a decade of efforts. One of the obstacles to the success is lack of an efficient and controllable coating methodology to achieve an biologically active surface with diverse function required for the healing process of surrounding tissues. In this study, we identified small adaptor peptides with titanium-binding property with a powerful screening methodology, phage display system. By five rounds of biopanning on Ti-6Al-4V discs, we isolated a clone of phage PT3 with high affinity to the surface of titanium, as verified by ELISA. We also conjugated PT3 at the amino terminus of hBMP-2 and expressed the fusion protein in E.coli. The identity of expressed protein was confirmed with mass spectrophotometry. It was then refolded and purified for further application in evaluation of the effect of immobilized growth factors on the healing process. This novel strategy may offer us a clinically efficient and easy-to-apply coating method in implant treatment.
huang, chi hung, and 黃濟鴻. "Generation of scFv against Thalidomide stereoisomer from phage display library." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/65105760400880058834.
Повний текст джерела國立中興大學
生物化學研究所
91
Thalidomide was first introduced in the 1950s as a sedative drug. Because it was deemed to be safe, it was prescribed for nausea and insomnia in pregnant women. However, it was found to cause the severe birth defects in children whose mothers had taken the drug in their first trimester of pregnancy. This was a terrible tragedy. Deepgoing research realized that one of thalidomide enantiomer possesses the desired therapeutic effect of quelling nausea, but its mirror compound produces fetal deformities. In this work, we synthesis thalidomide conjugated with BSA as an immunogen in order to produce related antibodies. After immunization, RT-PCR was performed and a series of primers was used to amplify splenic VH/VL genes. Then, amplified VH/VL were assembled to construct the antibody phage display library against thalidomide. The constructed antibody phage display library was being selected against immunogen after comfirming the diversity by BstNI restriction enzyme fingerprinting analysis. After three rounds of selection, we have isolated two phage clones with weak affinity against (+) or (-) thalidomide individually. Colony PCR and western blotting analysis showed these two clones bear full-length of scFv DNA fragment and are able to express scFv. Competitive ELISA assay also confirmed the specificity of these two scFvs against (+)-thalidomide or (-)-thalidomide respectively.
Hsia, Ko-Chiang, and 夏可強. "Isolation and characterization of lipase abzyme from phage displayed antibody library." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/mcyy49.
Повний текст джерела國立東華大學
生物技術研究所
94
Interactions of antigen and antibody resemble to some extents the recognition and specific binding between enzyme and substrate. For this reason, antibodies may have certain enzymatic activity and were named abzymes or catalytic antibodies. In the present work we aimed to get lipolytic abzymes through the selection from phage displayed antibody library. A transition state analog for selection of lipolytic abzymes was synthesized. The structure composed of 4-nitrophenyl activated phosphonate, the transition state analog, (TSA) of lipases/esterases, connected to a biotin moiety through a 15 carbon chain spacer. After several rounds of selection, we obtained six monoclonal phage particles capable of binding significantly to the TSA. However, no apparent enzyme activity was observed using the solution of these phage particles for lipase enzymatic assay. Then, we used strain HB2151 (K12 ara Δ(lac-proAB) thi/F' proA+B lacIq lacZΔM15) to express the soluble antibody fragment. Through the His-tag purification, the purified antibody protein was examined further for its binding capability to TSA and the possible enzymatic activity. The present method presents a more efficient and convenient means to find new abzyme. Catalytic antibodies may have the potential to replace current enzymes and may have very diverse application.
"Applications of phage-displayed antibody library for antibody discovery and engineering." Thesis, 2008. http://library.cuhk.edu.hk/record=b6074647.
Повний текст джерелаAttempts are made in the present study to extend three applications of the phage displayed antibody library in antibody discovery and engineering. Firstly, a CDR3-randomized phage-displayed scFv library was constructed from genomic DNA of mouse. Following biopanning, anti-peptide of mas oncoprotein scFvs were isolated and identified. These results illustrate the potential use of the genomic phage-displayed library for anti-peptide antibodies selection. Secondly, we described the isolation of anti-idiotypic scFvs against a chimeric anti-CD22 mAb from an immunized phage-displayed scFv library. The isolated anti-Id scFvs were able to capture the immune response of chimeric anti-CD22 mAb with high specificity. This reagent will enhance our understanding of the therapeutic mechanism of anti-CD22 mAb in non-Hodgkin's lymphoma treatment, and may be applied to probe the pharmacokinetics, tissue distribution, and modulation of anti-CD22 mAb in vivo.
In conclusion, we have attempted various approaches to identify specific anti-peptide scFvs, anti-idiotypic scFvs and passive anti-tumor scFvs. These results extend the applications of phage display technology in antibody discovery and engineering.
Our approach enables us to isolate selective and sensitive anti-idiotypic antibodies and could be exploited for other antibodies with clinical and biological applications. Thirdly, we profile a strategy to select and identify markers on tumor cell surface using phage-displayed antibodies from mice bearing xenograft tumor. Our data imply that passive antibodies in cancer patients may be obtained from the immune repertoire of cancer patients. Besides, we found a cell surface antigen was up-regulated more than 3-fold in mas-expressing cells. We further use the targeting antibody to construct a tumor endoprotease-activated immunotoxin.
Zhao, Qi.
Adviser: Wing-Tai Cheung.
Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3499.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2008.
Includes bibliographical references (leaves 227-250).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.
Tseng, Sy-Woei, and 曾斯偉. "Construction of phage-display peptide library for the screening of antibody epitope." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/04418277385769813527.
Повний текст джерела國立中興大學
分子生物學研究所
88
Abstract Phage display technology, consisting of the expression of target sequence in the surface of phage particles and the affinity selection, is a biological system that facilitates the cloning and rapid selection of peptides or proteins from large combinatorial libraries. Libraries are generated by cloning of a batch of DNA encoding millions of variants of certain ligands into the phage genome or phagemid as a fusion to the gene encoding one of the phage coat proteins. Upon expression, the fusion coat protein is incorporated into new phage particles that are assembled in the bacterium and consequently presented on the phage surface, with its genetic material residing within the phage particles. The purpose of this study is to construct a random peptide library containing both linear and conformational epitopes to serve as an “all purpose” peptide source for searching of disease-related peptides. To achieve this goal, EcoRI-HindIII DNA fragments encoding 45 amino acid residues, of which 36 have random sequences, were cloned into the corresponding sites of the modified pCANTAB5E plasmid and the resulted plasmids were transformed into E. coli TG1. The percentage of insert-containing colonies in the transformants was evaluated by PCR using primers flanking the insert DNAs. Approximately, 83% of the clones contained inserts. The expression of the insert gene was determined by colony immunoblot analysis of a portion of the original clones using the antibody specific for E-Tag which is located downstream of the insert. The number of clone capable of expression coincided with the percentage of the insert-containing clones. Finally, the diversity of the library was evaluated by sequencing of 10 randomly picked clones. The results indicated that all exhibited difference sequences. The diversity of the library was estimated to be 4.4 x 106 according to the above information,. To establish a system for identification of disease-related peptides, affinity selection was performed using sera from system lupus erythematosus patients.
Ying-HsiuLiu and 劉盈秀. "Selection and characterization of EV71 antibody by phage display human scFv antibody library." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/42895278044035675388.
Повний текст джерела國立成功大學
醫學檢驗生物技術學系碩博士班
101
Enterovirus 71 (EV71) which causes hand-foot-mouth disease (HFMD) and herpangina belongs to the Picornaviridae family with ss(+) RNA virus. In 1998, EV71 outbreak in Taiwan and resulted in 78 fatal cases of 405 severe complications related neurological or cardiopulmonary symptoms. EV71 has been threatened the children under 3 years of age in Asia-Pacific area. Vaccination of EV71 seems the most efficient to eradicate the infectious disease. However, the development of vaccines is still in clinical trials. Antibodies also develop for early detection and diagnosis to control and prevent EV71 transmission. Different antibody format can be expressed by phage display technology via genetic engineering. Here, we selected the phage clones to against EV71 from the human scFv antibody library via biopanning. Procedure of biopanning included binding the target, washing out unbound phage and amplification of binders. Phage clones in total 385 were random picked and grouped by RFLP with SalI restriction enzyme. Five groups were identified and sequenced the nucleotides. Sequence data were aligned with the international ImMunoGeneTics information system (IMGT). We found that all of five phage clones contain a variable heavy sequence. Phage clone 3 and 9 contained full length scFv nucleotide sequence but formed no functional scFv antibody in protein level. However, the phage clones 6 and 13 showed binding ability to against EV71 that revealed in ELISA results. In addition, the phage clone 6 and 13 can recognize EV71virus particles and VP2 protein in western blot. Phage clone 6 and 13 detected EV71 in RD cell can be observed by immunofluorescence assay (IFA). In this study, the data suggested that phage clone 6 and 13 from the human scFv antibody library can be applied for EV71 research or diagnosis.
Huang, Shih-Tsung, and 黃仕聰. "Development of Fully Human Anti-HDGF Antibody from Phage-Displayed Human Naïve ScFv Library." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/92wr6a.
Повний текст джерела國立中山大學
海洋生物科技博士學位學程
107
Hepatocellular carcinoma (HCC), a deadly disease worldwide. For advanced HCC therapy, Sorafenib is the only conventional drug, however, low response rate, drug resistance, and strong side effects had been reported in Sorafenib therapy. Therefore, investigation of novel therapeutic strategy is a critical issue. HDGF is a cancerous factor in various cancer types including HCC. To develop anti-HDGF neutralizing antibody, phage display technology provides an effective in vitro selection strategy to identify fully human antibody. In the biopanning, thirteen HDGF affinity phage clones had been identified from phage display human naïve scFv library using unique Fc fusion protein/protein G dynabeads method. After fully human IgG construction and production, these antibodies were further confirmed binding affinity using ELISA, immunofluorescence, and HDGF conditional knockdown Huh7 cell line. Among them, hmAb-7 exhibited multiple anti-cancer activity, including cell growth, colony formation, invasion and sphere formation in Huh7, HepG2, and SK-Hep1 cells. The anti-cancer effects of hmAb-7 might participated with downregulation of PI3K/AKT/mTOR signaling pathway. The hmAb-7 inhibits of invasion through MMP-2 down-regulation; sphere formation was eliminated through downregulation of cancer stemness genes. In continuously cultured Huh7 and HepG2 cells, these cells present higher colony and spheroid forming features, however, the cancerous behavior able to abolish through hmAb-7 treatment. In sorafenib resistant Huh7 cells, hmAb-7 presents colony and sphere inhibition ability. Furthermore, the Huh7 xenograft model in NOD/SCID mice was performed and the data indicated that hmAb-7 slightly inhibited tumor growth (p = 0.1091) and prolong the overall survival for 14 days. Although anti-HDGF hmAb-7 single treatments not significantly inhibit tumor growth in vivo, but based on the cancerous inhibition effects in vitro, combination therapy might provide an approach to enhance therapeutic effect. Furthermore, a HDGF sandwich ELISA platform with well linear range between 10-0.15625 ng/ml has been established and able to detect the HDGF level in human and rat serum. Anti-HDGF antibody was also used for extended applications, included in analgesic application and allow to treat cancer stemness and Sorafenib drug resistant Huh7 cells (Huh7-SR). Our finding indicated anti-HDGF hmAb-7 exhibited analgesic effect in rat chronic constrictive injury (CCI) model plus with Adenoviral HDGF gene delivery. Besides, hmAb-7 not only shown sphere and colony inhibitory effects in continuous spheroid cell cultured Huh7 and HepG2 cells, but also inhibited cancerous behaviors in Huh7-SR cells. Finally, adenovirus mediated anti-HDGF-scFv gene delivery suppressed the colony and sphere formation in both Huh7 and Huh7-SR cells. The adenovirus mediated scFv gene delivery provided a novel approach for anti-HDGF therapy and allowed for further basic studies.