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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Bibliographic Details
Published in:Frontiers in immunology 2021-12, Vol.12, p.747267-747267
Main Authors: Zhang, Hai-Nan, Xue, Jun-Biao, Wang, Aru Ze-Ling, Jiang, He-Wei, Merugu, Siva Bhararth, Li, Da-Wei, Tao, Sheng-Ce
Format: Article
Language:English
Subjects:
18A4Hu
Animals
Antibodies - immunology
Antibody Affinity - immunology
antibody affinity improvement
Antigens - immunology
Bacterial Proteins - genetics
Bacterial Proteins - immunology
directed evolution
DNA Polymerase I - immunology
DNA Polymerase I - metabolism
error-prone DNA polymerase I
Escherichia coli - immunology
Immunology
Lung Neoplasms - immunology
Melanoma - immunology
Mice
Mutation
protein-fragment complementation assay
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Summary: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 Assisted 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 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.
ISSN:1664-3224
1664-3224
DOI:10.3389/fimmu.2021.747267