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1
Austin, Eric B. "Human monoclonal antibodies." Thesis, University of Nottingham, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.276187.
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2
Plumpton, Christopher. "Monoclonal antibodies against phytochrome." Thesis, University of Cambridge, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.358677.
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3
Benjamin, Richard John. "Tolerance induction with monoclonal antibodies." Thesis, University of Cambridge, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.253988.
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4
Qin, Shi-Xin. "Transplantation tolerance with monoclonal antibodies." Thesis, University of Cambridge, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.305697.
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5
Heron, Andrew David. "The stability of monoclonal antibodies." Thesis, University of Glasgow, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.252169.
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6
Isaacs, John Dudley. "Improving serotherapy with monoclonal antibodies." Thesis, University of Cambridge, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.386115.
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7
Paudel, Subhash. "Shear thinning in monoclonal antibodies." Thesis, Kansas State University, 2016. http://hdl.handle.net/2097/32833.
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Master of Science<br>Department of Physics<br>Jeremy D. Schmit<br>Antibodies are large Y-shaped proteins which are used by immune system to identify and neutralize pathogens. Monoclonal antibody therapy is used to treat different patient conditions. There are problems associated with the manufacturability and deliverability of mAb solutions due to the viscous nature of the protein. The viscosity of antibody solutions increases with the increase in concentration and decreases with applied shear. We want to know why these behaviours are seen and to address this problem we have developed a theory describing the rapid viscosity increase with increasing concentration. We use the polymer theory to explain this behaviour. Here antibodies are treated as polymers. The length of the polymer depend on the aggregation. The reptation time increases approximately as the cubic power of size of aggregate (N³ ). We see the shear thinning behaviour is dependent on the Ab-Ab binding energy and find the relationship between the size of the aggregate
and the binding energy. We find aggregate size and morphology using several models for Ab-Ab interaction sites. We use the head to head binding (fAb-fAb binding) model to describe aggregation state in our viscosity theory. The size of the aggregate and hence the reptation time is captured by the binding energy. When the binding energy increases the zero shear viscosity increases and the reptation time decreases. Likewise when the binding energy decreases the zero shear viscosity decreases and the reptation time increases. We have yet to find the correct exponents for the shear thinning behaviour of different mAbs which would be our future work.
8
Ueda, Yasuji. "MONOCLONAL ANTIBODIES TO CHICK CRYSTALLINS." 京都大学 (Kyoto University), 1989. http://hdl.handle.net/2433/86412.
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9
Pathan, N. "Catalytic monoclonal antibodies: a review." Thesis(M.Phil.), CSIR-National Chemical Laboratory, Pune, 1990. http://dspace.ncl.res.in:8080/xmlui/handle/20.500.12252/2017.
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10
Alexandrovich, Susan K. "Characterization of monoclonal antibodies against digoxin /." Online version of thesis, 1987. http://hdl.handle.net/1850/10681.
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11
Mirza, Myriam. "Characterization of new CFTR monoclonal antibodies." Thesis, McGill University, 2009. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=66882.
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The available antibodies against CFTR are not sensitive enough to detect CFTR at endogenous or near endogenous levels making detection at native levels difficult. We raised two monoclonal antibodies, 22E8 and 23C5, against the R domain of human CFTR with the goal of identifying an antibody sensitive enough to detect CFTR in native airway cells. These antibodies were characterized for their ability to detect over-expressed as well as endogenous levels of CFTR in immunoblotting, immunoprecipitation and immunofluorescence. Their ability to detect CFTR was also compared with commercial antibodies M3A7 and 24-1. We show that 23C5 and 22E8 are more sensitive than the commercial antibodies and are able to detect CFTR in over-expressed and endogenous cells by immunoblotting. However, only 23C5 is able to immunoprecipitate CFTR and neither is able to detect CFTR in native airway cells by immunoblotting or are suitable for immunofluorescence. These antibodies will enable studies of CFTR biogenesis in endogenous cells.<br>A l'heure actuelle les anticorps dirigés contre la protéine CFTR ne sont pas suffisamment sensibles pour détecter cette protéine de facon endogène rendant ainsi l'étude de cette protéine difficile dans les tissus. Notre laboratoire a fabriqué deux anticorps monoclonaux , nommés 22E8 et 23C5, dirigés contre le domaine R de la protéine CFTR. L'abilité de ces anticorps à détecter l'expression de CFTR que ce soit de façon endogène ou lorsque la protéine est surexprimée a été testée à l'aide des techniques d'immunoblotting, d'immunoprécipitation et d'immunofluorescence. Afin de verifier leur sensibilité et leur capacité à détecter la protéine CFTR, ces anticorps ont été comparés aux anticorps M3A7 et 24-1 qui sont disponibles dans le commerce et connus pour détecter de facon optimale la protéine CFTR. Les resultats obtenus dans les lignées cellulaires à l'aide de la technique d'immunoblotting ont permis de montrer que les anticorps 23C5 et 22E8 sont plus sensibles que les anticorps commerciaux, de plus ils sont capables de détecter à la fois les protéines endogènes et sur-exprimées. Bien que l'anticorps 23C5 soit capable d'immunoprécipiter la protéine CFTR, aucun des deux anticorps n'a permis la detection de la protéine CFTR par immunoblotting dans les cellules de culture primaire. De plus, ces anticorps n'ont pas permis la detection de la protéine CFTR par immunofluorescence. Ainsi l'utilisation de ces anticorps nous donnera l'opportunité d'étudier la protéine CFTR dans les cellules l'exprimant de facon endogène afin de mieux comprendre sa regulation et son traffic.
12
Noble, Philip W. "Characterisation of anti-glycan monoclonal antibodies." Thesis, University of Nottingham, 2011. http://eprints.nottingham.ac.uk/12071/.
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The aims of this thesis are to establish the therapeutic value of two anti-glycan mAbs produced in-house, to develop an immunisation protocol with the aim of improving the immunogenicity tumour-associated glycolipids with the intention of producing therapeutically valuable mAbs and to determine the implication of a mAb with the ability to induce apoptosis in colorectal cancer. The anti-glycan mAbs 692/29 and 505/4 have previously been produced in-house and this study aimed to determine their fine specificity using a glycan array. 692/29 displayed binding predominantly to Lewis b as well as Lewis y-containing glycans. 505/4 was discovered to bind to sialyl Lewis a as well as sialyl di-Lewis a, with no cross-reactivity with other blood group antigens. This was compared to other anti-Lewis mAbs, with differences in specificity being observed. Characterisation of 505/4 mAb distribution showed binding to 80% of colorectal tumours and low levels of binding to normal tissues by IHC, suggesting it may be therapeutically useful. This thesis aimed to assess the ability of 505/4 and 692/29 to meditate immune mediated and non-immune mediated cell death as well as to determine whether non-immune-mediated cell death would be a desirable therapeutic property. Resistance to apoptosis is one of the hallmarks of cancer cells and mAbs stimulating apoptosis may not be very effective. Alternatively, cancer cells are driven to initiate apoptosis by genomic and other aberrations thus if pro-apoptotic pathways are stimulated these cells may be more susceptible to death than normal cells. To investigate the significance of apoptosis in cancer a large tissue microarray of colorectal tumours was assessed for apoptosis and its relationship to patient prognosis. Cleaved caspase-3 is a good marker of apoptosis as it is the executioner caspase for both the extrinsic and intrinsic pathways. Immunohistochemical analysis of colorectal tumour samples revealed that a high expression of cleaved caspase-3 in tumour was associated with good prognosis in colorectal cancer. This suggested that some tumours were still susceptible to apoptotic death but some are resistant and an alternative mechanism of cell death may be an advantage in these tumours. High expression of cleaved caspase-3 in the tumour-associated stroma was also an independent marker of good prognosis in colorectal cancer. This may be because apoptosis of the tumour-associated stroma reduces the level of pro-tumour signals originating from tumour-associated immune cells and stromal cells. As the tumour microenvironment can act in an immunosuppressive and pro-tumour manner, the ability of a mAb to induce direct cell death without the need for effector cells or complement would be an advantage. Lewis y and Lewis b are blood group antigens commonly overexpressed on the surface of a range of cancers. Characterisation of effector functions of 505/4 and 692/29 demonstrated that both mAbs have the ability to mediate apoptosis by antibody dependent cellular cytotoxicity, complement dependent cytotoxicity and cause direct cell death in an oncosis-like manner. Comparison with other anti-Lewis mAbs demonstrated that a number of anti-Lewis mAbs can induce direct cell death independently of apoptosis. Thus, they could effectively target apoptotic sensitive and resistant colorectal cancers. Tumours aberrantly express glycolipids and these molecules may be involved in a number of cellular pathways. In addition a large proportion of anti-glycan mAbs, including 505/4 and 692/29 in this thesis, have displayed the ability to induce direct cell death. Therefore this thesis aimed to develop an immunisation protocol capable of increasing the immunogenicity of tumour-associated glycolipid for the production of anti-tumour glycolipid mAbs directed against ovarian cancer. This study suggests that the incorporation of tumour glycolipid into liposomes and their immunisation along with the iNKT cell adjuvant α-galactosylceramide, elicits an anti-tumour glycolipid immune response, which can yield IgG mAbs capable of binding a high proportion of ovarian cancers. In summary, this thesis confirmed specificity of 692/29 to Lewis y and Lewis b and 505/4 to sialyl Lewis a and sialyl di-Lewis a. Furthermore, this thesis demonstrated a promising tissue distribution of 505/4 in vitro. Characterisation of mAb effector functions suggest that both Lewis y and sialyl Lewis a directed mAbs have the ability to cause direct cell death, independently of apoptosis in antigen positive cells, as well as the ability to cause immune-mediated cell death. This may be an important factor in the immune-suppressive tumour microenvironment. Furthermore, this thesis provides the basis for the production of new anti-glycolipid antibodies that may also be able to induce direct cell death.
13
Thanh, Le Thiet. "Exon-specific monoclonal antibodies against dystrophin." Thesis, University of Salford, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.261661.
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14
Watson, Nigel. "Monoclonal antibodies to human immunoglobulin allotypes." Thesis, University of Strathclyde, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.304897.
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15
Ortlepp, Susan. "Leucocyte integrin activation by monoclonal antibodies." Thesis, Open University, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.359976.
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16
Holdsworth, Mary Louise. "Characterisation of phytochrome using monoclonal antibodies." Thesis, University of Leicester, 1987. http://hdl.handle.net/2381/35466.
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Characterisation of phytochrome using monoclonal antibodies Mary L. Holdsworth Native oat phytochrome has been purified to homogeneity and used to produce a panel of monoclonal antibodies (mAbs). Selection of mAbs followed early screening against native phytochrome by ELISA, and SDS-denatured phytochrome by "mini" western blotting. Six mAbs which recognised SDS-denatured phytochrome were mapped using proteolytically derived fragments of phytochrome and subsequent immunoblotting. LAS 31 and 33 map to the 6 kDa NH2-terminus and LAS 35 and 41 map to the adjacent 4 kDa sub-NH2- terminal domain. LAS 11 maps to the 64 kDa- chromophore-bearing domain and LAS 32 maps to between 74 and 88 kDa from the NH2-terminus on the COOH- terminal-half of the molecule. A novel protocol for the mapping of conformation-specific mAbs has been developed and used to assign LAS 21, 34 and 42 to the 64 kDa-chromophore-bearing domain. Determination of differential affinities towards Pr and Pfr demonstrated that LAS 42 exhibited a higher affinity for Pfr, LAS 31, 33, 34 and 35 exhibited a higher affinity for Pr. LAS 41 discriminates absolutely against Pfr. LAS 41 has therefore facilitated:- (i) the purification of PfrP, i.e. Pfr which is free of contaminating Pr, (ii) the development of an ELISA for phytochrome photoequilibrium, (iii) the first direct experimental evidence that phytochrome can exist as a stable heterodimer in vitro and (iv) an appraisal of ELISA protocols for determining differential affinities of mAbs towards Pr and Pfr. Spectral analyses of phytochrome in the presence of mAbs have underlined the importance of the 6 kDa NH2-terminus in the maintenance of the spectral integrity of the molecule but have also indicated that the 4 kDa sub-NH2-terminal domain also interacts with the chromophore. Cross reactivity studies amongst phytochrome from monocots and dicots demonstrate that the epitopes recognised by LAS 11, 31, 33, 35 and 41 are not highly conserved. However, LAS 32 recognises phytochrome from every plant species tested, and is therefore recognising a highly conserved region on the molecule.
17
Giorno, Caterina [Verfasser]. "Glycoengineering of Monoclonal Antibodies / Caterina Giorno." Konstanz : Bibliothek der Universität Konstanz, 2010. http://d-nb.info/1020366117/34.
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18
Yeung, Douglas Edward. "Characterization of six monoclonal antibodies against the Minute Virus of Mice NS-1 protein, and the use of one in the immunoaffinity purification of NS-1 expressed in insect cells." Thesis, University of British Columbia, 1990. http://hdl.handle.net/2429/29405.
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Six mouse monoclonal antibodies have been isolated which react against a bacterial fusion protein containing amino acids 364 to 623 of the NS-1 protein of the prototype strain of the Minute Virus of Mice (MVMp). All six were found to be of the IgG class of antibodies; five being IgG₁ and the sixth being IgG₂[formula omitted]. By immunoblot analyses, these antibodies all recognize an 83 kDa protein found only in MVM-infected mouse fibroblast cells, leading to the assumption that they are all NS-1 specific. Further evidence for this assumption is obtained from indirect immunofluorescence studies showing all but one of the mAbs react against a nuclear protein found in MVM-infected cells.
The epitopes of the antibodies were mapped using carboxy-terminal deleted bacterial fusion proteins derived from the plasmid encoding the original antigen. For the six monoclonal antibodies, four distinct epitopes were found (A - D). Three were clustered in a 16 amino acid region near the carboxy-terminal of the bacterial fusion protein, while the fourth was slightly more toward the amino-terminal side. Competition ELISAs against a 25 amino acid NS-1 specific peptide confirmed the mapping of the A epitope recognized by the CE10 and AC6 monoclonal antibodies.
Also in this thesis, the characterization of a NS-1 fusion protein and a non-fused NS-1 protein expressed in insect cells by recombinant baculoviruses is also described. The latter, a full-length NS-1 protein designated NS-1[formula omitted]ⅽ, was found to be an 84 kDa cytoplasmic protein. This protein was immunoprecipitated by all six monoclonal antibodies. A CE10 monoclonal antibody immunoaffinity column was employed in the single-step purification of NS-1 [formula omitted]c from insect cells. Four elution methods (alkaline, peptide, 6M guanidinium, and acid) were examined and the best purification was obtained using the acid elution.<br>Medicine, Faculty of<br>Biochemistry and Molecular Biology, Department of<br>Graduate
19
Ohlin, Mats. "Human monoclonal antibody technology a tool to investigate human antibody repertoires /." Lund : Dept. of Immunotechnology, Lund University, 1992. http://catalog.hathitrust.org/api/volumes/oclc/39693827.html.
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20
Evans, Rachael Yvonne. "The production of anti-idiotopic antibodies to monoclonal anti-RhD antibodies." Thesis, Lancaster University, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.274194.
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21
Chen, Desheng, and chen desheng@deakin edu au. "Development of monoclonal antibodies against Vibrio pathogens." Deakin University. Department of Biological Science, 1991. http://tux.lib.deakin.edu.au./adt-VDU/public/adt-VDU20080626.140825.
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Monoclonal antibodies were developed against pathogenic vibrios for use in rapid identification in disease situations of humans, fish and shellfish. Of the 12 fusions performed using V. alginolyticus, V. anguillarum, V. carchariae, V. cholerae, V. damsela, V. furnissii, V. harveyi, V. ordalii, V. parahaemolyticus and V. vulnificus, a total of 102 hybridomas were obtained. Based on cross-reactivity of a wide range of Vibrio strains and other gram-negative bacteria, three broad types of monoclonal antibodies were found. The three categories were: (1) ones that were species-specific or specific to a particular surface antigen, (2) a large number that reacted with several Vibrio species, and (3) three that reacted with most Vibrio strains but no other gram-negative bacteria.
Each species-specific monoclonal antibody only recognized its corresponding Vibrio species and was used for identifying unknown species, confirming diagnosis of clinical isolates. In addition, several monoclonal antibodies only cross-reacted with similar Vibrio species, e.g. V. parahaemolyticus and V. alginolyticus which share a common H-antigen. Monoclonal antibodies reacting with several Vibrio species were not of particular use in diagnostic situations. Three monoclonal antibodies of the last group did not react with other genera of the family Vibrionaceae, namely Aeromonas, Photobacterium and Plesiomonas nor a wide range of gram-negative enteric bacteria. These data indicated the existence of an antigenic surface determinant common to Vibrio species. One monoclonal reacted with the heat-stable antigenic determinants on the cell surface as v as lipopolysaccharide extracted from all the vibrios studied, thus making it useful for large- scale screening of acute infections of vibrios.
In a blind test, seven Vibrio species, isolated from 6 marine and a freshwater source were identified by two laboratories using phenetic tests. Results of immunotyping using monoclonals, three of seven were diagnosed as the same species, another three were designated as Vibrio species but could not be classified further due to the library not having the corresponding monoclonal, and one was diagnostically questionable. Two further tests were carried out. An unknown Vibrio formalin-fixed isolated from diseased marine animal was identified as
V. parahaemolyticus by ELISA and FITC. Clinical human isolates of V. alginolyticus, V. parahaemolyticus and V. vulnificus were confirmed by monoclonals.
Australian isolates of V. anguillarum appeared to be mostly of serotype O1. monoclonals raised to V. anguillarum AFHRL 1 reacted with only serotype O1 from Denmark but also most Australian isolates.
All vibrios pathogenic to fish and shellfish, i.e. V. anguillarum, V. ordalii, V. alginolyticus, V. carchariae, V. cholerae, V. damsela, V. harveyi, V. parahaemolyticus and V. vulnificus, were used for attachment studies to fish cells using phase contrast and FITC-immunofluorescence microscopy. Of these vibrios, V. anguillarum, V. ordalii and V. perahaemolyticus, were found to adhere to different cells and tissues of rainbow trout while others did not appear to attach. However, attachment was inhibited by monoclonal antibodies specific to only these three vibrios.
Lipopolysaccharide is well known as being a contributing factor in pathogenicity of gram-negative bacteria. PAGE electrophoresis of extracted LPS from 9 strains covering 6 Vibrio species showed the presence of a common 15,000 D fragment. This fragment was verified by immunoblotting with a genus-specific monoclonal antibody (i.e. F11P411F) recognizing nearly all vibrios. The common LPS fragment was separated and used to raise polyclonal antisera in mouse which reacted strongly with LPS itself, live as well as sodium azide-killed vibrios, but not with other gram-negative bacteria. This raised the possibility of developing vaccine from Vibrio LPS.
Monoclonal antibodies developed in the present study enabled rapid identification of a number of pathogenic Vibrio species. There is still further work to produce monoclonal antibodies against additional vibrios that are probably pathogenic. These included V. fluvialis, V. hollisae, V. metschnikovii, V. minicus, V. salmonella and V. tubiashii. Together the application will be of significance in clinical diagnostic work, in the monitoring of vibriosis in fish farms and in quarantine.
22
Bell, Ian Martin. "Monoclonal antibodies as catalysts for cationic cyclisations." Thesis, University of Cambridge, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.387041.
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23
Storey, E. "Monoclonal antibodies to merozoites of Plasmodium falciparum." Thesis, University of Oxford, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.371577.
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24
Banbury, David N. "New monoclonal antibodies to visualise vesicular compartments." Thesis, University of Oxford, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.259782.
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25
Klutz, Stephan [Verfasser]. "Continuous processing of monoclonal antibodies / Stephan Klutz." München : Verlag Dr. Hut, 2016. http://d-nb.info/1115549731/34.
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26
Honey, C. R. "Immunosuppression with monoclonal antibodies in neural transplantation." Thesis, University of Oxford, 1990. http://ora.ox.ac.uk/objects/uuid:ea39dc7a-4ada-4c21-8cef-4649cb322646.
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27
Huang, Ling. "Investigation of soya globulins using monoclonal antibodies." Thesis, University of East Anglia, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.302022.
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28
Anderson, J. "Investigations on bluetongue virus using monoclonal antibodies." Thesis, Open University, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.374892.
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The main aim of the thesis was the production and characterisation of monoclonal antibodies (Mabs) against bluetongue virus (BTV) and the application of such antibodies to the improvement of BTV diagnosis. The suitability of the indirect ELISA for the detection of antibodies to BTV was examined along with the published protocols for the purification of BTV ELISA antigen. When sera were examined from animals which had experienced either BTV vaccine or any other tissue-culture derived vaccine, host-cell protein contaminants in the BTV ELISA antigen reacted with anti-BHK cell antibodies to give false positive results. None of the published protocols completely removed host-cell protein contaminants. As a solution to these problems monoclonal antibodies were produced to BTV and characterised as to their reactivity against a panel of BTV antigen preparations. They were further characterised by competitive binding assays, and SDS-PAGE analysis of the various BTV preparations with which they reacted optimally. One Mab (3-17-A3), directed against BTV p7, proved suitable for use in a blocking ELISA for the detection of antibodies to all 22 serotypes of BTV. The blocking ELISA proved both more sensitive and more specific than the currently used agar gel precipitin test, and has been adopted as the screening test before importation of semen and embryos into the United Kingdom. A further group of antibodies (characterised by Mab F58) directed against BTV polypeptide NS1, used in a blocking assay detected anti-BTV tubule antibodies. This allows detection of BTV replication in infected animals, and may be used to discriminate between BTV infected animals and animals vaccinated with inactivated BTV vaccine. This work demonstrated the previously unreported BTV-specific nature of BTV NS1. Serum-free suspension culture of the hybridomas and purification of the Mab from tissue-culture supernatant was also developed.
29
Jones, Christine Ann. "Monoclonal antibodies in the study of neuropeptides." Thesis, Imperial College London, 1991. http://hdl.handle.net/10044/1/46848.
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30
Yeates, Peter Gregory. "Tumour imaging using hCG-specific monoclonal antibodies." Thesis, The University of Sydney, 1989. https://hdl.handle.net/2123/26188.
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The aim of this thesis was to radiolabel two different anti-hCG antibodies with 111-indium, and to determine their specific tumour uptake and tissue biodistribution into choriocarcinoma xenografts in nude mice against 111-indium and 67-gallium citrate over 72 hours. Comparative radioimmunoscintigraphy was performed to determine the optimal imaging post injection for visualisation of tumour sites. Data manipulation using dual isotope computerised subtraction in mouse phantom models was evaluated as means of improving tumour delineation in low tumour contrast studies.
31
Chen, Jianqing. "Radiolabeling and biotinylation of internalizing monoclonal antibody chimeric BR96 potential use for extracorporeal immunoadsorption with enhanced tumor radioactivity retention of iodine, indium and rhenium /." Lund : Lund University, the Jubileum Institute, Dept. of Radiation Physics, 1997. http://catalog.hathitrust.org/api/volumes/oclc/39725797.html.
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32
Xu, Wenbin. "Studies of antigenic relationships among spotted fever group rickettsiae by monoclonal antibodies." Aix-Marseille 2, 1997. http://www.theses.fr/1997AIX20665.
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33
Hills, Anna E. "Control of monoclonal antibody N-glycosylation." Thesis, University of Kent, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.344101.
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34
Sun, Ping. "Studies of sperm antigens using specific monoclonal antibodies." Thesis, University of British Columbia, 1987. http://hdl.handle.net/2429/28405.
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This thesis work is focused on studies of sperm antigen: its purification and antifertility effect, its "internal image" by use of anti-idiotypic antibodies, and its capacity to induce antibodies of IgA class.
Mouse sperm acrosomal antigens that react with a monoclonal antibody generated against mouse spermatozoa, MS 207, were purified by immunoaffinity chromatography from soluble fraction of mouse testis homogenates. Analyzed by HPLC and SDS gel electrophoresis in the presence of reducing agent, the purified sperm antigens were shown to have a molecular weight of 600 to 700 KDa. It was shown to be glycoproteins with carbohydrate content of 23%. The purified mouse sperm antigen was used to isoimmunize female mice. After successive immunizations, antisera of high titers were raised and were shown to react specifically with mouse and human sperm, but not with any mouse somatic tissues as judged from results of immunofluorescent assay and Ouchterlony's double immunodiffusion. The antisera were shown to crossreact with human sperm at the acrosomal region. By using mouse in vitro fertilization experiments and human sperm penetration assay with zona-free hamster ova, the antisera were shown to exert strong inhibitory effects on fertilization. Another part of this thesis work was directed to generating anti-idiotypic antibody against a monoclonal sperm antibody MS 204. MS 204 was previously shown to react with a conformational epitope of sperm antigens. Anti-MS 204 antiserum was raised in a rabbit. A series of affinity chromatographic columns were applied to purify anti-Fab fragment of MS 204 (AId 204). The obtained Aid 204 possessed weak antigenicity in the mouse. After successive immunizations with Aid 204, the isoimmune sera against AId 204 were shown to crossreact with acrosomal region of mouse sperm, a pattern similar to that of MS 204. A partial inhibitory effect on in vitro fertilization was observed. The results indicate that the "internal image" of the specific sperm antigen reactive with MS 204 exists in AId 204. This could be a new strategy to obtain conformational epitope of a given antigen and may serve as the alternative of contraceptive vaccines.<br>Medicine, Faculty of<br>Obstetrics and Gynaecology, Department of<br>Graduate
35
Tsim, Karl Wah-Keung. "Chick acetylcholinesterase : purification, molecular properties and monoclonal antibodies." Thesis, University of Cambridge, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.236054.
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Acetylcholinesterase (AChE, E.C. 3.1.1.7) from 1-day chick brain was enriched over 2,000-fold by N-methylacridinium affinity column. Using this preparation as immunogen, two monoclonal antibodies (mAbs), 1E2 and 3D10, were isolated. Both mAbs react with all molecular forms of AChE from chick but not with butyrylcholinesterases (BuChE, E.C. 3.1.1.8). The mAb, 1E2, was used to immunopurify chick brain globular and muscle asymmetric AChE to homogeneity. The purified brain AChE showed a specific activity of 2,200 U/mg of protein, and it appeared to be a hydrophobic tetramer with a subunit mass of 105 kDa. The purified muscle asymmetric AChE has a specific activity of 1,100 U/mg of protein, it exhibits catalytic and inhibition properties characteristic of both AChE and BuChE and contains three distinct subunits with an apparent size of 110 kDa, 72 kDa and 58 kDa in the ratio 2:2:1. The discovery of an AChE/BuChE hybrid asymmetric form has been further supported by: (1) the identification of active site properties of AChE in the 110kDa subunit and of BuChE in the 72-kDa subunit, (2) the purification and precipitation of both activities by a BuChE-specific mAb (7D11), and (3) the evidence that all subunits are bound in the asymmetric form by disulphide bonds. The 58-kDa subunit is the only one that is sensitive to digestion with purified collagenase; it carries the collagenous 'tail'.
36
Gilliland, Lisa Kim. "The development of bispecific monoclonal antibodies for therapy." Thesis, University of Cambridge, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.278214.
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37
Watkins, I. D. "Monoclonal antibodies to Legionella pneumophila and related organisms." Thesis, University of Oxford, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.354879.
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38
Tellides, George. "Immunosupression monoclonal antibodies to rat lymphocyte activation antigens." Thesis, University of Oxford, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.236271.
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Chia, T. W. H. "The use of monoclonal antibodies for glycoside hydrolysis." Thesis, University of Cambridge, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.597597.
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Since 1986, a wide range of reactions have been catalysed by monoclonal antibodies, many with good rate enhancements, high regio- and stereo-selectivities and high substrate specificities. As antibodies can be elicited against practically any molecule of interest, this new class of antibody molecules, commonly known as abzymes, offers a unique way of generating tailor-made, enzyme-like catalysts. We propose to investigate the possibility of using monoclonal antibodies to catalyse the hydrolysis of glycosides. Glycosides are important components of many biological processes especially in glycoproteins which make up the cell walls of bacteria and the outer coats of viruses. Catalytic antibodies raised may serve as therapeutic agents to selectively hydrolyse such carbohydrate coats. In addition, they can be used to probe the mechanisms of glycoside hydrolysis. In our initial attempts, we synthesised nojirimycin-like transition state analogues (these compounds have an extra methylene group at the C2 position) which when protonated, copies a similar charge in the transition state. Although we were successful in synthesising the benzyl-protected compound and incorporating the uv-detectable nitrophenyl molecule at C2 (to mimic the transition state of analogous nitrophenyl glucosides), the deprotection step, unfortunately, removed the nitrophenyl group too. We next attempted to synthesise an amidine analogue which would mimic both the charge and conformation of the transition state. However, the synthesis of the key intermediate, D-gluconolactam, was unsuccessful. Another chemical was then synthesised. Hydroximo-lactones are good inhibitors of β-glucosidases and could also serve as transition state analogues. Incorporation of the dinitrophenyl group followed by deprotection of the acetyl-protecting groups provided the desired product without affecting the uv-group. Attempts to attach a linker molecule (this is to minimise the response directed to the protein when raising monoclonal antibodies) directly to this hapten were unsuccessful and the linker was ultimately introduced in a protected form although this esterification step was not regioselective. Deprotection of the <i>tert</i>-butyl group of one isomer provided the hapten.
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Mason, Caroline Margaret. "Characterisation of intestinal M cells using monoclonal antibodies." Thesis, University of Newcastle Upon Tyne, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.260952.
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Perera, W. Shermal. "Binding and molecular characterisation of monoclonal RhD antibodies." Thesis, University of Aberdeen, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.342706.
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Monoclonal RhD antibodies (anti-RhD) may potentially be used in preventing Rh alloimmunisation of women delivering RhD-positive babies, and thereby preventing haemolytic disease of the newborn (HDN). Binding and molecular analyses were performed on a range of monoclonal anti-RhDs to understand the interaction between the antibody and antigen. A flow cytometry (FCM) assay system was developed to analyse the binding of the anti-RhD to human group O R<sub>1</sub>R<sub>2</sub> red blood cells (RBC). A panel of 30 anti-RhD preparations were studied using this assay and Vmax (maximum binding) and KD (equilibrium constant) were determined for each antibody. The antibodies were categorised into 3 groups (low, medium and high binders) according to their Vmax. Furthermore, the Vmax was converted to the number of antibody molecules bound per RBC (NMBR) by using a correlation curve generated from running parallel FCM and radioiodination assays (RIA). Scatchard analysis of the RIA data indicated that the total NMBR for O R<sub>1</sub>R<sub>2</sub> RBC was 27,300 antigen sites/cell. Molecular analysis involved cloning and sequencing of 11 anti-RhD. Heavy chains (HC) preferentially used gene segments from the VH3 and VH4 families, and kappa chain (κ LC) usage was restricted to DPK9. Four sets of antibodies, showed restricted D gene segments (encode part of the HCDR3) which indicated possible importance for epitope specificity. Analysis of V gene sequence indicated that common VH and VL pairings were found used by the medium binders. The high and low binders had unique VH and VL pairings, although the high binders also showed greater somatic mutations from their respective germline genes. It was concluded that the fit of the antibody to the RhD antigen is dependent on both the VH and VL usage and pairing, and that the precise epitope specificity of these antibodies may require HCDR3 interaction.
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Duguid, I. G. M. "Prevention of corneal graft rejection with monoclonal antibodies." Thesis, University of Aberdeen, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.387460.
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This thesis aims to place corneal allograft rejection in the context of general transplantation immunology, examine the role of lymphocyte subsets in the rejection process and consider the potential application of monoclonal antibody therapy in clinical corneal graft rejection. The literature relating to the current clinical practice of corneal grafting, with particular reference to corneal allograft rejection, is reviewed in chapter 1 to present the extent of the problem. Chapter 2 then reviews the mechanisms of allograft rejection from the literature of transplantation immunology, much of which has arisen from studies of kidney, heart, pancreatic islets and liver in animal models. The materials and methods are described in detail in chapter 3, and only the relevant experimental design is detailed in the Materials and Methods sections of the succeeding chapters. The experimental mouse model of transplanting corneal tissue into the renal subcapsular is evaluated in chapter 4, demonstrating that isografts survive indefinitely whereas allografts are rejected typically by 30 days. Pretransplant sensitisation decreased allograft survival time to 10 days. Immunohistochemistry demonstrated the presence of CD4<sup>+</sup> and CD8<sup>+</sup> lymphocytes and macrophages at the rejection site. Heterotopic corneal graft recipients were then treated with various monoclonal antibody regimes. Chapter 5 demonstrates that allograft survival can be increased by either anti-CD4 or anti-CD8 therapy, providing near total depletion of the respective lymphocyte subset is achieved. Xenograft rejection is shown to depend on mainly CD4<sup>+</sup> lymphocytes in chapter 6, with no benefit being found of depleting the CD8<sup>+</sup> subset in addition. A mild immunosuppressive effect of anti-Vβ8 monoclonal antibody is demonstrated and discussed in chapter 7. The final chapter discusses these results in the light of recent, related work in other transplant systems, and presents a case for a trial of intracameral pan-T-cell monoclonal antibody treatment.
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De, Silva M. G. "Human monoclonal antibodies in the study of diabetes." Thesis, University of Surrey, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.233146.
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Bell, Graham Thomas. "Studies on the production of human monoclonal antibodies." Thesis, University of Edinburgh, 1988. http://hdl.handle.net/1842/19241.
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Buffetto, Fanny. "Generation of new monoclonal antibodies to rhamnogalacturonan fragments." Nantes, 2014. https://archive.bu.univ-nantes.fr/pollux/show/show?id=85b86558-be8b-4c38-ae82-702e14160fa0.
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Les pectines sont largement présentes dans la lamelle moyenne et la paroi primaire des dicotylédones. Elles sont composées de différents domaines structuraux liés de façon covalente. Les rhamnogalacturonanes II et les rhamnogalacturonanes I sont les domaines pectiques les plus complexes. Le rhamnogalacturonane II est formé d'une chaîne principale d'homogalacturonane sur laquelle sont branchées 5 chaînes latérales contenant de nombreux oses rares. Le rhamnoglaladuronane·I possède un squelette·hétérogène de rhamnoses et d'acides galacturoniques. Ce domaine contient des ramifications constituées d' arabinoses et de galactoses. Pour localiser ces structures in planta, la microscopie en immunofluorescence est une technique·sensible. . . Les immu nomarquages sont réalisés par le biais d' anticorps reconnaissant des structures très spécifiques. Certains anticorps spécifiques des pectines sont déjà disponibles, cependant la productiorn de nouvelles sondes est nécessaire. Pour générer de nouveaux anticorps, des oligosaccharides ,issus de rhamnogalacturonane II et rhamnogalacturonane I ont été isolés. La préparation de ces oligosaccharides a révélé une grande diversité de motifs structuraux composant ces domaines. Les immunisations menées avec les différents oligosaccharides ont permis la génération d'un nouvel amticorps contre·les chaînes latérales des rhamnogalacturonane I. Cet anticorps reconnaît un motif structural, jusqu'alors très peu décrit dans la littérature:, mais présent dans les parois cellulaires riches en galactanes. Cette structure in planta a été localisée dans le tubercule de pomme de terre ainsi que dans les jeunes racines d’Arabidopsis. La fonction de cette structure reste encore à détermi ner<br>Pectins are widely present in middle lamellae and primary cell walls of dicots. They are composed of different covalently linked structural domains. Rhamnogalacturonan II and rhamnogalacturonan I are the most complex pectic domains. Rhamnogalacturonan II is composed of a homogalacturonan backbone, which carries five different side chains containing rare sugars. Rhamnogalacturonan I has a heterobackbone of rhamnoses and galacturonic acids. This domain contains linear or branched arabinose ‐ and galactose ‐ rich chains. To localize these structures in planta, immunofluorescence microscopy is a sensitive technique. Immunolabelling is performed with antibodies, recognizing specific structures. Some antibodies, which label pectins are already available. However, the production of new probes is required. To generate new antibodies, oligosaccharides from rhamnogalacturonan II and rhamnogalacturonan I were purified. Structural analyses revealed the high diversity of motifs present in rhamnogalacturonan I. Immunizations carried out with the different oligosaccharides have enabled the generation of a new monoclonal antibody binding to rhamnogalacturonan I side chains. This antibody recognizes a molecular motif present in galactan rich sources, which has been poorly described in literature. This structure was localized in planta, in potato tuber and in roots of seedling Arabidopsis. However, function of this structure remains to be determined
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Eastwood, David Geoffrey Douglas. "Immunotoxicology of the therapeutic monoclonal antibody TGN1412." Thesis, St George's, University of London, 2015. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.676898.
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Having passed all pre-clinical safety testing, the superagonistic anti -CD28 therapeutic monoclonal antibody (mAb ) TGN 1412, intended for the treatment of rheumatoid arthritis and B-cell chronic lymphocytic leukaemia, was approved by German and UK regulatory authorities for first-in-man Phase One clinical trial. Shortly after infusion, all six healthy trial volunteers suffered unexpected and profound systemic pro-inflammatory cytokine release, later termed a 'cytokine storm,' causing multi-organ failure. This unexpected and near fatal cytokine release syndrome (CRS) publically highlighted the failure of current pre-clinical safety testing procedures, emphasising an urgent need for novel cytokine release assays (CRAs) capable of predicting adverse properties of therapeutic mAbs. A wet coat mAb immobilisation approach, developed here, has proven predictive of clinical outcome and would have anticipated TGN1412 immunotoxicity in man. This approach IS now being widely applied by the pharmaceutical industry and contract research organisations (CROs). Comparative studies, testing TGN 1412 against a panel , of therapeutic mAbs, identified a unique mechanism of TGNl412-driven cytokine release. Substantial concentrations of the cytokine lL-2 were subsequently found to be a hallmark for the cytokine storm observed in-vivo, signifYing IL-2 as a TGN1412-like response biomarker. Multiple pro-inflammatory cytokine release was also shown to be principally effector memory T cell (T EM) derived in man. Human and macaque comparative immunophenotyping crucially identified macaque T EM cells as lacking CD28 expression, explaining pre-clinical animal model testing failures. A more physiologically relevant aqueous phase co-culture assay using monocyte-derived dendritic cells is also shown capable of eliciting a TGN1412-like cytokine release profile equivalent to that detected in-vivo and in-vitro using wet coat immobilisation; implying presentation in-vivo likely involved dendritic cells. This thesis describes the most likely mechanisms of action responsible for TGN1412 immunotoxicology in man and provides a plausible explanation for the pre-clinical safety testing failures, findings vital to the fields of immunomodulatory therapeutic mAb development and immunotoxicology.
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Kalofonos, Haralabos. "Radiolabelled monoclonal antibodies for tumour immunoscintigraphy and biodistribution studies." Thesis, Imperial College London, 1990. http://hdl.handle.net/10044/1/46376.
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Joyce, Christopher Francis. "The production and functional assessment of IGA monoclonal antibodies." Thesis, Imperial College London, 1990. http://hdl.handle.net/10044/1/46374.
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Brush, Bradley Alan. "Characterization of monoclonal antibodies to rubella virus structural proteins." Thesis, University of British Columbia, 1989. http://hdl.handle.net/2429/27400.
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Monoclonal antibodies (MAbs), due to their intrinsic specificity to single
determinants, are capable of defining the antigenic and structural properties
of viruses. A panel of twenty-five monoclonal antibodies, the result of
fusions between splenocytes from Balb/c mice immunized with RA 27/3 vaccine
strain rubella virus and myeloma cell line NS1X63Ag8.653, were characterized by
radioimmunoprecipitation of rubella virus structural proteins, IgG
sub-classification, haemagglutination inhibition (HI) and virus neutralization
(Nt) of rubella virus (RV).
The antigen specificities of thirteen of twenty-five MAbs were identified
by immunoprecipitation of [³⁵S]-methionine labelled virus.
All showed reactivity to the E1 envelope glycoprotein of either M33 wild type, or RA 27/3 vaccine strain virus, or both. The remainder precipitated multiple viral proteins or none at all. These trials required numerous alterations in the composition of buffers to achieve unambiguous results.
IgG subclassification was performed on hybridoma cell culture supernatants or purified ascites fluid by the Ouchterlony double immunodiffusion technique, with all immunoglobulins typed as IgG₁, IgG[sub 2a] or IgG[sub 2b], but with no
cases of IgG₃ or IgM secretors.
Inhibition of rubella virus agglutination of chick erythrocytes by nine of the monoclonal antibodies gave haemagglutination inhibition titres of greater than 8 with the highest titre at 16384.
The panel of monoclonal antibodies was tested for its capacity to neutralize both M33 and RA 27/3 strains of rubella virus, with and without the presence of complement. To detect rubella plaque formation exclusively, an immunocytochemical method of detecting rubella virus proteins on cell surfaces was developed. Plaque reduction experiments showed that ten of the MAbs inhibited the cytopathic effect of rubella virus to some extent. Five MAbs neutralized M33 RV without complement present, one showing a four-fold increase in titre upon its addition, and one MAb, negative in the absence of complement displayed a positive titre in its presence. Three MAbs showed neutralizing activity toward RA 27/3 with no observed effect upon the addition of complement; two MAbs showed a shift from negativity to neutralizing titres. Of the ten MAbs that were active, four reacted with both rubella strains. The capacity of the monoclonal antibodies to neutralize rubella virus was found to be independent of their ability to inhibit the haemagglutinin of the virus. Therefore the epitopes for virus neutralization and haemagglutination appear to be different.<br>Medicine, Faculty of<br>Pathology and Laboratory Medicine, Department of<br>Graduate
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Hunter, Nicole Marie. "A system for the production of human monoclonal antibodies." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape8/PQDD_0006/MQ46024.pdf.
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