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A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies

The classical `knob-into-holes' (KIH) strategy (knob(T366Y)/hole (Y407T)) has successfully enhanced the heterodimerization of a bispecific antibody (BsAb) resulting in heterodimer formation up to 92% of protein A (ProA)-purified protein pool. However, it does not show high efficiency for every...

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Autores principales: Wang, Bo, Lin, Jun, Hoag, Matthew R, Wright, Meredith, Ma, Mingjun, Cai, Wenyan, Gallolu Kankanamalage, Sachith, Liu, Yue
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9413979/
https://www.ncbi.nlm.nih.gov/pubmed/36042698
http://dx.doi.org/10.1093/abt/tbac019
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author Wang, Bo
Lin, Jun
Hoag, Matthew R
Wright, Meredith
Ma, Mingjun
Cai, Wenyan
Gallolu Kankanamalage, Sachith
Liu, Yue
author_facet Wang, Bo
Lin, Jun
Hoag, Matthew R
Wright, Meredith
Ma, Mingjun
Cai, Wenyan
Gallolu Kankanamalage, Sachith
Liu, Yue
author_sort Wang, Bo
collection PubMed
description The classical `knob-into-holes' (KIH) strategy (knob(T366Y)/hole (Y407T)) has successfully enhanced the heterodimerization of a bispecific antibody (BsAb) resulting in heterodimer formation up to 92% of protein A (ProA)-purified protein pool. However, it does not show high efficiency for every BsAb. KIH was initially applied to a CD20/CD3 BsAb. After in silico modeling, two additional new mutations, S354Y in knob-heavy chain (HC) and Q347E in hole-HC, together with KIH named `ETYY', were introduced in the Fc. The CD20/CD3 BsAb hybrid only represented ~ 50% of the ProA-purified protein pool when KIH was applied. With ETYY, the percentage of CD20/CD3 hybrid increased to 93.8%. CD20/CD3-v4b (containing ETYY) retains the original activity of the BsAb at both Fab and Fc regions, and also shows good developability. These results indicate that the computer-aided novel ETYY design has the potential to improve the development of next-generation BsAbs with higher yields and simpler purification.
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spelling pubmed-94139792022-08-29 A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies Wang, Bo Lin, Jun Hoag, Matthew R Wright, Meredith Ma, Mingjun Cai, Wenyan Gallolu Kankanamalage, Sachith Liu, Yue Antib Ther Original research article The classical `knob-into-holes' (KIH) strategy (knob(T366Y)/hole (Y407T)) has successfully enhanced the heterodimerization of a bispecific antibody (BsAb) resulting in heterodimer formation up to 92% of protein A (ProA)-purified protein pool. However, it does not show high efficiency for every BsAb. KIH was initially applied to a CD20/CD3 BsAb. After in silico modeling, two additional new mutations, S354Y in knob-heavy chain (HC) and Q347E in hole-HC, together with KIH named `ETYY', were introduced in the Fc. The CD20/CD3 BsAb hybrid only represented ~ 50% of the ProA-purified protein pool when KIH was applied. With ETYY, the percentage of CD20/CD3 hybrid increased to 93.8%. CD20/CD3-v4b (containing ETYY) retains the original activity of the BsAb at both Fab and Fc regions, and also shows good developability. These results indicate that the computer-aided novel ETYY design has the potential to improve the development of next-generation BsAbs with higher yields and simpler purification. Oxford University Press 2022-08-01 /pmc/articles/PMC9413979/ /pubmed/36042698 http://dx.doi.org/10.1093/abt/tbac019 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Antibody Therapeutics. All rights reserved. For Permissions, please email: journals.permissions@oup.com https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Original research article
Wang, Bo
Lin, Jun
Hoag, Matthew R
Wright, Meredith
Ma, Mingjun
Cai, Wenyan
Gallolu Kankanamalage, Sachith
Liu, Yue
A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
title A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
title_full A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
title_fullStr A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
title_full_unstemmed A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
title_short A novel IgG Fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
title_sort novel igg fc by computer-aided design enhances heavy-chain heterodimerization in bi- or trispecific antibodies
topic Original research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9413979/
https://www.ncbi.nlm.nih.gov/pubmed/36042698
http://dx.doi.org/10.1093/abt/tbac019
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