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Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher
Biparatopic fragment antibodies can overcome deficiencies in avidity of conventional antibody fragments. Here, we describe a technology for generating biparatopic antibodies through two-step targeting using a pair of polypeptides, SpyTag and SpyCatcher, that spontaneously react to form a covalent bo...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6976922/ https://www.ncbi.nlm.nih.gov/pubmed/31993343 http://dx.doi.org/10.1016/j.btre.2020.e00418 |
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author | Akiba, Hiroki Takayanagi, Kensuke Kusano-Arai, Osamu Iwanari, Hiroko Hamakubo, Takao Tsumoto, Kouhei |
author_facet | Akiba, Hiroki Takayanagi, Kensuke Kusano-Arai, Osamu Iwanari, Hiroko Hamakubo, Takao Tsumoto, Kouhei |
author_sort | Akiba, Hiroki |
collection | PubMed |
description | Biparatopic fragment antibodies can overcome deficiencies in avidity of conventional antibody fragments. Here, we describe a technology for generating biparatopic antibodies through two-step targeting using a pair of polypeptides, SpyTag and SpyCatcher, that spontaneously react to form a covalent bond between antibody fragments. In this method, two antibody fragments, each targeting different epitopes of the antigen, are fused to SpyTag and to SpyCatcher. When the two polypeptides are serially added to the antigen, their proximity on the antigen results in covalent bond formation and generation of a biparatopic antibody. We validated the system with purified recombinant antigen. Results in antigen-overexpressing cells were promising although further optimization will be required. Because this strategy results in high-affinity targeting with a bipartite molecule that has considerably lower molecular weight than an antibody, this technology is potentially useful for diverse applications. |
format | Online Article Text |
id | pubmed-6976922 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-69769222020-01-28 Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher Akiba, Hiroki Takayanagi, Kensuke Kusano-Arai, Osamu Iwanari, Hiroko Hamakubo, Takao Tsumoto, Kouhei Biotechnol Rep (Amst) Research Article Biparatopic fragment antibodies can overcome deficiencies in avidity of conventional antibody fragments. Here, we describe a technology for generating biparatopic antibodies through two-step targeting using a pair of polypeptides, SpyTag and SpyCatcher, that spontaneously react to form a covalent bond between antibody fragments. In this method, two antibody fragments, each targeting different epitopes of the antigen, are fused to SpyTag and to SpyCatcher. When the two polypeptides are serially added to the antigen, their proximity on the antigen results in covalent bond formation and generation of a biparatopic antibody. We validated the system with purified recombinant antigen. Results in antigen-overexpressing cells were promising although further optimization will be required. Because this strategy results in high-affinity targeting with a bipartite molecule that has considerably lower molecular weight than an antibody, this technology is potentially useful for diverse applications. Elsevier 2020-01-11 /pmc/articles/PMC6976922/ /pubmed/31993343 http://dx.doi.org/10.1016/j.btre.2020.e00418 Text en © 2020 Published by Elsevier B.V. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Akiba, Hiroki Takayanagi, Kensuke Kusano-Arai, Osamu Iwanari, Hiroko Hamakubo, Takao Tsumoto, Kouhei Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher |
title | Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher |
title_full | Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher |
title_fullStr | Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher |
title_full_unstemmed | Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher |
title_short | Generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using SpyTag and SpyCatcher |
title_sort | generation of biparatopic antibody through two-step targeting of fragment antibodies on antigen using spytag and spycatcher |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6976922/ https://www.ncbi.nlm.nih.gov/pubmed/31993343 http://dx.doi.org/10.1016/j.btre.2020.e00418 |
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