Cargando…
Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life
We generated an anti-albumin antibody, CA645, to link its Fv domain to an antigen-binding fragment (Fab), thereby extending the serum half-life of the Fab. CA645 was demonstrated to bind human, cynomolgus, and mouse serum albumin with similar affinity (1–7 nM), and to bind human serum albumin (HSA)...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5058626/ https://www.ncbi.nlm.nih.gov/pubmed/27315033 http://dx.doi.org/10.1080/19420862.2016.1185581 |
_version_ | 1782459268490854400 |
---|---|
author | Adams, Ralph Griffin, Laura Compson, Joanne E. Jairaj, Mark Baker, Terry Ceska, Tom West, Shauna Zaccheo, Oliver Davé, Emma Lawson, Alastair DG. Humphreys, David P. Heywood, Sam |
author_facet | Adams, Ralph Griffin, Laura Compson, Joanne E. Jairaj, Mark Baker, Terry Ceska, Tom West, Shauna Zaccheo, Oliver Davé, Emma Lawson, Alastair DG. Humphreys, David P. Heywood, Sam |
author_sort | Adams, Ralph |
collection | PubMed |
description | We generated an anti-albumin antibody, CA645, to link its Fv domain to an antigen-binding fragment (Fab), thereby extending the serum half-life of the Fab. CA645 was demonstrated to bind human, cynomolgus, and mouse serum albumin with similar affinity (1–7 nM), and to bind human serum albumin (HSA) when it is in complex with common known ligands. Importantly for half-life extension, CA645 binds HSA with similar affinity within the physiologically relevant range of pH 5.0 – pH 7.4, and does not have a deleterious effect on the binding of HSA to neonatal Fc receptor (FcRn). A crystal structure of humanized CA645 Fab in complex with HSA was solved and showed that CA645 Fab binds to domain II of HSA. Superimposition with the crystal structure of FcRn bound to HSA confirmed that CA645 does not block HSA binding to FcRn. In mice, the serum half-life of humanized CA645 Fab is 84.2 h. This is a significant extension in comparison with < 1 h for a non-HSA binding CA645 Fab variant. The Fab-HSA structure was used to design a series of mutants with reduced affinity to investigate the correlation between the affinity for albumin and serum half-life. Reduction in the affinity for MSA by 144-fold from 2.2 nM to 316 nM had no effect on serum half-life. Strikingly, despite a reduction in affinity to 62 µM, an extension in serum half-life of 26.4 h was still obtained. CA645 Fab and the CA645 Fab-HSA complex have been deposited in the Protein Data Bank (PDB) with accession codes, 5FUZ and 5FUO, respectively. |
format | Online Article Text |
id | pubmed-5058626 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-50586262016-10-24 Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life Adams, Ralph Griffin, Laura Compson, Joanne E. Jairaj, Mark Baker, Terry Ceska, Tom West, Shauna Zaccheo, Oliver Davé, Emma Lawson, Alastair DG. Humphreys, David P. Heywood, Sam MAbs Report We generated an anti-albumin antibody, CA645, to link its Fv domain to an antigen-binding fragment (Fab), thereby extending the serum half-life of the Fab. CA645 was demonstrated to bind human, cynomolgus, and mouse serum albumin with similar affinity (1–7 nM), and to bind human serum albumin (HSA) when it is in complex with common known ligands. Importantly for half-life extension, CA645 binds HSA with similar affinity within the physiologically relevant range of pH 5.0 – pH 7.4, and does not have a deleterious effect on the binding of HSA to neonatal Fc receptor (FcRn). A crystal structure of humanized CA645 Fab in complex with HSA was solved and showed that CA645 Fab binds to domain II of HSA. Superimposition with the crystal structure of FcRn bound to HSA confirmed that CA645 does not block HSA binding to FcRn. In mice, the serum half-life of humanized CA645 Fab is 84.2 h. This is a significant extension in comparison with < 1 h for a non-HSA binding CA645 Fab variant. The Fab-HSA structure was used to design a series of mutants with reduced affinity to investigate the correlation between the affinity for albumin and serum half-life. Reduction in the affinity for MSA by 144-fold from 2.2 nM to 316 nM had no effect on serum half-life. Strikingly, despite a reduction in affinity to 62 µM, an extension in serum half-life of 26.4 h was still obtained. CA645 Fab and the CA645 Fab-HSA complex have been deposited in the Protein Data Bank (PDB) with accession codes, 5FUZ and 5FUO, respectively. Taylor & Francis 2016-06-17 /pmc/articles/PMC5058626/ /pubmed/27315033 http://dx.doi.org/10.1080/19420862.2016.1185581 Text en Published with license by Taylor & Francis Group, LLC © 2016 UCB http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. |
spellingShingle | Report Adams, Ralph Griffin, Laura Compson, Joanne E. Jairaj, Mark Baker, Terry Ceska, Tom West, Shauna Zaccheo, Oliver Davé, Emma Lawson, Alastair DG. Humphreys, David P. Heywood, Sam Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life |
title | Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life |
title_full | Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life |
title_fullStr | Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life |
title_full_unstemmed | Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life |
title_short | Extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin Fv domain: An investigation into the correlation between affinity and serum half-life |
title_sort | extending the half-life of a fab fragment through generation of a humanized anti-human serum albumin fv domain: an investigation into the correlation between affinity and serum half-life |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5058626/ https://www.ncbi.nlm.nih.gov/pubmed/27315033 http://dx.doi.org/10.1080/19420862.2016.1185581 |
work_keys_str_mv | AT adamsralph extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT griffinlaura extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT compsonjoannee extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT jairajmark extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT bakerterry extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT ceskatom extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT westshauna extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT zaccheooliver extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT daveemma extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT lawsonalastairdg extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT humphreysdavidp extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife AT heywoodsam extendingthehalflifeofafabfragmentthroughgenerationofahumanizedantihumanserumalbuminfvdomainaninvestigationintothecorrelationbetweenaffinityandserumhalflife |