EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem

Antibodies are one of the most important groups of biomolecules for both clinical and basic research and have been developed as potential therapeutics. Affinity is the key feature for biological activity and clinical efficacy of an antibody, especially of therapeutic antibodies, and thus antibody af...

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Autores principales:	Hai-nan Zhang, Jun-biao Xue, Aru Ze-ling Wang, He-wei Jiang, Siva Bhararth Merugu, Da-wei Li, Sheng-ce Tao
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	antibody affinity improvement directed evolution 18A4Hu error-prone DNA polymerase I protein-fragment complementation assay Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/9733a6ddde754c66b8518f3ccfab7729
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spelling	oai:doaj.org-article:9733a6ddde754c66b8518f3ccfab77292021-12-03T05:07:49ZEASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem1664-322410.3389/fimmu.2021.747267https://doaj.org/article/9733a6ddde754c66b8518f3ccfab77292021-12-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fimmu.2021.747267/fullhttps://doaj.org/toc/1664-3224Antibodies are one of the most important groups of biomolecules for both clinical and basic research and have been developed as potential therapeutics. Affinity is the key feature for biological activity and clinical efficacy of an antibody, especially of therapeutic antibodies, and thus antibody affinity improvement is indispensable and still remains challenging. To address this issue, we developed the E. coliAssisted Speed affINity-maturation Evolution SyStem (EASINESS) for continuous directed evolution of Ag–Ab interactions. Two key components of EASINESS include a mutation system modified from error-prone DNA polymerase I (Pol I) that selectively mutates ColE1 plasmids in E. coli and a protein–protein interaction selection system from mDHFR split fragments. We designed a GCN4 variant which barely forms a homodimer, and during a single round of evolution, we reversed the homodimer formation activity from the GCN4 variant to verify the feasibility of EASINESS. We then selected a potential therapeutic antibody 18A4Hu and improved the affinity of the antibody (18A4Hu) to its target (ARG2) 12-fold in 7 days while requiring very limited hands-on time. Remarkably, these variants of 18A4Hu revealed a significant improved ability to inhibit melanoma pulmonary metastasis in a mouse model. These results indicate EASINESS could be as an attractive choice for antibody affinity maturation.Hai-nan ZhangJun-biao XueAru Ze-ling WangHe-wei JiangSiva Bhararth MeruguDa-wei LiSheng-ce TaoFrontiers Media S.A.articleantibody affinity improvementdirected evolution18A4Huerror-prone DNA polymerase Iprotein-fragment complementation assayImmunologic diseases. AllergyRC581-607ENFrontiers in Immunology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	antibody affinity improvement directed evolution 18A4Hu error-prone DNA polymerase I protein-fragment complementation assay Immunologic diseases. Allergy RC581-607
spellingShingle	antibody affinity improvement directed evolution 18A4Hu error-prone DNA polymerase I protein-fragment complementation assay Immunologic diseases. Allergy RC581-607 Hai-nan Zhang Jun-biao Xue Aru Ze-ling Wang He-wei Jiang Siva Bhararth Merugu Da-wei Li Sheng-ce Tao EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem
description	Antibodies are one of the most important groups of biomolecules for both clinical and basic research and have been developed as potential therapeutics. Affinity is the key feature for biological activity and clinical efficacy of an antibody, especially of therapeutic antibodies, and thus antibody affinity improvement is indispensable and still remains challenging. To address this issue, we developed the E. coliAssisted Speed affINity-maturation Evolution SyStem (EASINESS) for continuous directed evolution of Ag–Ab interactions. Two key components of EASINESS include a mutation system modified from error-prone DNA polymerase I (Pol I) that selectively mutates ColE1 plasmids in E. coli and a protein–protein interaction selection system from mDHFR split fragments. We designed a GCN4 variant which barely forms a homodimer, and during a single round of evolution, we reversed the homodimer formation activity from the GCN4 variant to verify the feasibility of EASINESS. We then selected a potential therapeutic antibody 18A4Hu and improved the affinity of the antibody (18A4Hu) to its target (ARG2) 12-fold in 7 days while requiring very limited hands-on time. Remarkably, these variants of 18A4Hu revealed a significant improved ability to inhibit melanoma pulmonary metastasis in a mouse model. These results indicate EASINESS could be as an attractive choice for antibody affinity maturation.
format	article
author	Hai-nan Zhang Jun-biao Xue Aru Ze-ling Wang He-wei Jiang Siva Bhararth Merugu Da-wei Li Sheng-ce Tao
author_facet	Hai-nan Zhang Jun-biao Xue Aru Ze-ling Wang He-wei Jiang Siva Bhararth Merugu Da-wei Li Sheng-ce Tao
author_sort	Hai-nan Zhang
title	EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem
title_short	EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem
title_full	EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem
title_fullStr	EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem
title_full_unstemmed	EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem
title_sort	easiness: e. coli assisted speedy affinity-maturation evolution system
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/9733a6ddde754c66b8518f3ccfab7729
work_keys_str_mv	AT hainanzhang easinessecoliassistedspeedyaffinitymaturationevolutionsystem AT junbiaoxue easinessecoliassistedspeedyaffinitymaturationevolutionsystem AT aruzelingwang easinessecoliassistedspeedyaffinitymaturationevolutionsystem AT heweijiang easinessecoliassistedspeedyaffinitymaturationevolutionsystem AT sivabhararthmerugu easinessecoliassistedspeedyaffinitymaturationevolutionsystem AT daweili easinessecoliassistedspeedyaffinitymaturationevolutionsystem AT shengcetao easinessecoliassistedspeedyaffinitymaturationevolutionsystem
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EASINESS: E. coli Assisted Speedy affINity-maturation Evolution SyStem

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