Rapid and robust antibody Fab fragment crystallization utilizing edge-to-edge beta-sheet packing.

Tytuł:: Rapid and robust antibody Fab fragment crystallization utilizing edge-to-edge beta-sheet packing.
Autorzy:: Lieu R; Biotechnology Discovery Research, Applied Molecular Evolution, Eli Lilly and Company, San Diego, CA, United States of America.
Antonysamy S; Discovery Chemistry Research and Technologies, Eli Lilly and Company Corporate Center, Indianapolis, IN, United States of America.
Druzina Z; Discovery Chemistry Research and Technologies, Eli Lilly and Company Corporate Center, Indianapolis, IN, United States of America.
Ho C; Biotechnology Discovery Research, Applied Molecular Evolution, Eli Lilly and Company, San Diego, CA, United States of America.
Kang NR; Biotechnology Discovery Research, Applied Molecular Evolution, Eli Lilly and Company, San Diego, CA, United States of America.
Pustilnik A; Discovery Chemistry Research and Technologies, Eli Lilly and Company Corporate Center, Indianapolis, IN, United States of America.
Wang J; Discovery Chemistry Research and Technologies, Eli Lilly and Company Corporate Center, Indianapolis, IN, United States of America.
Atwell S; Biotechnology Discovery Research, Applied Molecular Evolution, Eli Lilly and Company, San Diego, CA, United States of America.
Źródło:: PloS one [PLoS One] 2020 Sep 11; Vol. 15 (9), pp. e0232311. Date of Electronic Publication: 2020 Sep 11 (Print Publication: 2020).
Typ publikacji:: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.
Język:: English
Imprint Name(s):: Original Publication: San Francisco, CA : Public Library of Science
MeSH Terms:: Immunoglobulin Fab Fragments/*chemistry
Immunoglobulin kappa-Chains/*chemistry
Amino Acid Sequence ; Animals ; CHO Cells ; Cricetulus ; Crystallization ; Crystallography, X-Ray ; Humans ; Protein Conformation, beta-Strand ; Rabbits
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Grant Information:: P30 GM124169 United States GM NIGMS NIH HHS
Substance Nomenclature:: 0 (Immunoglobulin Fab Fragments)
0 (Immunoglobulin kappa-Chains)
Entry Date(s):: Date Created: 20200911 Date Completed: 20201027 Latest Revision: 20211006
Update Code:: 20240105
PubMed Central ID:: PMC7485759
DOI:: 10.1371/journal.pone.0232311
PMID:: 32915778
: Czasopismo naukowe

Pełny tekst

Antibody therapeutics are one of the most important classes of drugs. Antibody structures have become an integral part of predicting the behavior of potential therapeutics, either directly or as the basis of modeling. Structures of Fab:antigen complexes have even greater value. While the crystallization and structure determination of Fabs is easy relative to many other protein classes, especially membrane proteins, broad screening and optimization of crystalline hits is still necessary. Through a comprehensive review of rabbit Fab crystal contacts and their incompatibility with human Fabs, we identified a small secondary structural element from the rabbit light chain constant domain potentially responsible for hindering the crystallization of human Fabs. Upon replacing the human kappa constant domain FG loop (HQGLSSP) with the two residue shorter rabbit loop (QGTTS), we dramatically improved the crystallization of human Fabs and Fab:antigen complexes. Our design, which we call "Crystal Kappa", enables rapid crystallization of human fabs and fab complexes in a broad range of conditions, with less material in smaller screens or from dilute solutions.
Competing Interests: RL, SA, ZD, CH, NRK, AP, JW and SA are, or were at the time of the work described here, employees of Eli Lilly & Co. Eli Lilly & Co. provided support in the form of salaries for the authors and research funding. Eli Lilly & Co. also approved the publication of this work. Eli Lilly & Co. has filed a patent application related to the work described here. The employment of the authors by Eli Lilly & Co does not alter our adherence to PLOS ONE policies on data sharing and materials.

Erratum in: PLoS One. 2021 Oct 5;16(10):e0258412. (PMID: 34610050)

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