{"id":{"repo_id":"westminster","oai_identifier":"oai:westminsterresearch.westminster.ac.uk:90088"},"canonical_url":"https://search.dev.ndltd.org/etd/westminster/oai:westminsterresearch.westminster.ac.uk:90088","repository":{"repo_id":"westminster","name":"University of Westminster","base_url":"https://westminsterresearch.westminster.ac.uk/oai2"},"display":{"title":"In vitro selection and characterisation of human anti-HIV-1 antibody fragments","abstract":"Generation of neutralising antibodies with broad specificity would be one of the effective approaches to control HIV-1 spread. It is clear that a method that allows rapid generation of neutralising antibodies is needed. This project aims at developing a novel approach to rapidly access human anti-HIV-1 antibodies in vitro by using ribosome display and selection from DNA libraries of HIV-1 patients. Two single-chain antibody libraries (M325 and K530) were constructed from two HIV-1 long-term non-progressors, whose sera showed cross-neutralising activities against various HIV-1 strains across a range of clades. In each library, total RNA was extracted from blood of each donor and used to synthesise cDNA. Families of 4 κ light chains, 9 λ light chains and 8 heavy chains were generated by using RT-PCR amplification. These fragments were then assembled with all possible combinatorial pairs to form diversified repertories in the form of VL-link-VH-partial CH. Both libraries were subjected to ribosome display for in vitro selection of functional antibodies. Ribosome display is a cell-free technique used to generate proteins that can bind to an immobilised antigen. During this process, the translated proteins are associated with their mRNAs, enabling a simultaneous selection of functional proteins and their gene. The employment of ribosome display facilitated rapid screening of two large libraries against recombinant gp120 (generated from patient K530).","abstract_html":"Generation of neutralising antibodies with broad specificity would be one of the effective approaches to control HIV-1 spread. It is clear that a method that allows rapid generation of neutralising antibodies is needed. This project aims at developing a novel approach to rapidly access human anti-HIV-1 antibodies in vitro by using ribosome display and selection from DNA libraries of HIV-1 patients. Two single-chain antibody libraries (M325 and K530) were constructed from two HIV-1 long-term non-progressors, whose sera showed cross-neutralising activities against various HIV-1 strains across a range of clades. In each library, total RNA was extracted from blood of each donor and used to synthesise cDNA. Families of 4 κ light chains, 9 λ light chains and 8 heavy chains were generated by using RT-PCR amplification. These fragments were then assembled with all possible combinatorial pairs to form diversified repertories in the form of VL-link-VH-partial CH. Both libraries were subjected to ribosome display for in vitro selection of functional antibodies. Ribosome display is a cell-free technique used to generate proteins that can bind to an immobilised antigen. During this process, the translated proteins are associated with their mRNAs, enabling a simultaneous selection of functional proteins and their gene. The employment of ribosome display facilitated rapid screening of two large libraries against recombinant gp120 (generated from patient K530).","abstract_has_math":false,"creators":["Tang, J."],"institution":"University of Westminster","degree_name":null,"degree_level":"PhD thesis","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011","date_published":"2011","updated_at":"2026-07-24T06:02:11Z","subjects":[],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["oai:westminsterresearch.westminster.ac.uk:90088"],"render_values":[{"text":"oai:westminsterresearch.westminster.ac.uk:90088","href":null,"code":true}]}]},"links":{"outbound_url":"https://doi.org/10.34737/90088","outbound_label":"DOI","outbound_source":"dc:identifier.doi"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Tang, J."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011"]},{"key":"dc:date.issued","label":"Date","values":["2011"]},{"key":"dc:publisher","label":"Institution","values":["University of Westminster"]},{"key":"dc:publisher.department","label":"Dc Publisher Department","values":["School of Life Sciences"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Westminster"]},{"key":"dc:relation","label":"Dc Relation","values":["https://westminsterresearch.westminster.ac.uk/item/90088/in-vitro-selection-and-characterisation-of-human-anti-hiv-1-antibody-fragments"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://westminsterresearch.westminster.ac.uk/item/90088/in-vitro-selection-and-characterisation-of-human-anti-hiv-1-antibody-fragments"]},{"key":"dc:type","label":"Dc Type","values":["Thesis or dissertation"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["PhD thesis"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["oai:westminsterresearch.westminster.ac.uk:90088"]},{"key":"dc:identifier.doi","label":"DOI","values":["https://doi.org/10.34737/90088"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://westminsterresearch.westminster.ac.uk/download/47aefbe9e5c65c9a6edc088376be2078875a21c336c4859d1a3146e816795aa8/15283519/Jie_TANG.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Generation of neutralising antibodies with broad specificity would be one of the effective approaches to control HIV-1 spread. It is clear that a method that allows rapid generation of neutralising antibodies is needed. This project aims at developing a novel approach to rapidly access human anti-HIV-1 antibodies in vitro by using ribosome display and selection from DNA libraries of HIV-1 patients. Two single-chain antibody libraries (M325 and K530) were constructed from two HIV-1 long-term non-progressors, whose sera showed cross-neutralising activities against various HIV-1 strains across a range of clades. In each library, total RNA was extracted from blood of each donor and used to synthesise cDNA. Families of 4 κ light chains, 9 λ light chains and 8 heavy chains were generated by using RT-PCR amplification. These fragments were then assembled with all possible combinatorial pairs to form diversified repertories in the form of VL-link-VH-partial CH. Both libraries were subjected to ribosome display for in vitro selection of functional antibodies. Ribosome display is a cell-free technique used to generate proteins that can bind to an immobilised antigen. During this process, the translated proteins are associated with their mRNAs, enabling a simultaneous selection of functional proteins and their gene. The employment of ribosome display facilitated rapid screening of two large libraries against recombinant gp120 (generated from patient K530)."]},{"key":"dc:description.abstract","label":"Abstract","values":["Generation of neutralising antibodies with broad specificity would be one of the effective approaches to control HIV-1 spread. It is clear that a method that allows rapid generation of neutralising antibodies is needed. This project aims at developing a novel approach to rapidly access human anti-HIV-1 antibodies in vitro by using ribosome display and selection from DNA libraries of HIV-1 patients. Two single-chain antibody libraries (M325 and K530) were constructed from two HIV-1 long-term non-progressors, whose sera showed cross-neutralising activities against various HIV-1 strains across a range of clades. In each library, total RNA was extracted from blood of each donor and used to synthesise cDNA. Families of 4 κ light chains, 9 λ light chains and 8 heavy chains were generated by using RT-PCR amplification. These fragments were then assembled with all possible combinatorial pairs to form diversified repertories in the form of VL-link-VH-partial CH. Both libraries were subjected to ribosome display for in vitro selection of functional antibodies. Ribosome display is a cell-free technique used to generate proteins that can bind to an immobilised antigen. During this process, the translated proteins are associated with their mRNAs, enabling a simultaneous selection of functional proteins and their gene. The employment of ribosome display facilitated rapid screening of two large libraries against recombinant gp120 (generated from patient K530)."]},{"key":"dc:title","label":"Title","values":["In vitro selection and characterisation of human anti-HIV-1 antibody fragments"]}]}],"canonical_facts":{"dc:creator":["Tang, J."],"dc:date":["2011"],"dc:date.issued":["2011"],"dc:description":["Generation of neutralising antibodies with broad specificity would be one of the effective approaches to control HIV-1 spread. It is clear that a method that allows rapid generation of neutralising antibodies is needed. This project aims at developing a novel approach to rapidly access human anti-HIV-1 antibodies in vitro by using ribosome display and selection from DNA libraries of HIV-1 patients. Two single-chain antibody libraries (M325 and K530) were constructed from two HIV-1 long-term non-progressors, whose sera showed cross-neutralising activities against various HIV-1 strains across a range of clades. In each library, total RNA was extracted from blood of each donor and used to synthesise cDNA. Families of 4 κ light chains, 9 λ light chains and 8 heavy chains were generated by using RT-PCR amplification. These fragments were then assembled with all possible combinatorial pairs to form diversified repertories in the form of VL-link-VH-partial CH. Both libraries were subjected to ribosome display for in vitro selection of functional antibodies. Ribosome display is a cell-free technique used to generate proteins that can bind to an immobilised antigen. During this process, the translated proteins are associated with their mRNAs, enabling a simultaneous selection of functional proteins and their gene. 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These fragments were then assembled with all possible combinatorial pairs to form diversified repertories in the form of VL-link-VH-partial CH. Both libraries were subjected to ribosome display for in vitro selection of functional antibodies. Ribosome display is a cell-free technique used to generate proteins that can bind to an immobilised antigen. During this process, the translated proteins are associated with their mRNAs, enabling a simultaneous selection of functional proteins and their gene. The employment of ribosome display facilitated rapid screening of two large libraries against recombinant gp120 (generated from patient K530)."],"dc:identifier":["oai:westminsterresearch.westminster.ac.uk:90088"],"dc:identifier.doi":["https://doi.org/10.34737/90088"],"dc:identifier.uri":["https://westminsterresearch.westminster.ac.uk/download/47aefbe9e5c65c9a6edc088376be2078875a21c336c4859d1a3146e816795aa8/15283519/Jie_TANG.pdf"],"dc:publisher":["University of Westminster"],"dc:publisher.department":["School of Life Sciences"],"dc:publisher.institution":["University of Westminster"],"dc:relation":["https://westminsterresearch.westminster.ac.uk/item/90088/in-vitro-selection-and-characterisation-of-human-anti-hiv-1-antibody-fragments"],"dc:relation.isreferencedby":["https://westminsterresearch.westminster.ac.uk/item/90088/in-vitro-selection-and-characterisation-of-human-anti-hiv-1-antibody-fragments"],"dc:title":["In vitro selection and characterisation of human anti-HIV-1 antibody fragments"],"dc:type":["Thesis or dissertation"],"dc:type.qualificationlevel":["PhD thesis"]},"updated_at":"2026-07-24T06:02:11Z"}