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

DOI: 10.1371/journal.pone.0232311 DOI Help

Authors: Ricky Lieu (Eli Lilly and Company) , Stephen Antonysamy (Eli Lilly and Company) , Zhanna Druzina (Eli Lilly and Company) , Carolyn Ho (Eli Lilly and Company) , N. Rebecca Kang (Eli Lilly and Company) , Anna Pustilnik (Eli Lilly and Company) , Jing Wang (Eli Lilly and Company) , Shane Atwell (Eli Lilly and Company)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Plos One , VOL 15

State: Published (Approved)
Published: September 2020
Diamond Proposal Number(s): 22808

Open Access Open Access

Abstract: 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.

Journal Keywords: Crystal structure; Crystals; Crystallization; Rabbits; Crystal structure refinement; Crystallization seeding; Diffraction; Membrane protein crystallization

Subject Areas: Biology and Bio-materials, Medicine

Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

Other Facilities: Beamline 31-ID at Advanced Photon Source; Beamline ALS-502 at Advanced Light Source

Added On: 23/09/2020 10:28


Discipline Tags:

Life Sciences & Biotech Health & Wellbeing Drug Discovery Structural biology

Technical Tags:

Diffraction Macromolecular Crystallography (MX)