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Title: Protein crystallization with microseed matrix screening: application to human germline antibody Fabs

The power of microseed matrix screening is demonstrated in the crystallization of a panel of antibody Fab fragments. The crystallization of 16 human antibody Fab fragments constructed from all pairs of four different heavy chains and four different light chains was enabled by employing microseed matrix screening (MMS). In initial screening, diffraction-quality crystals were obtained for only three Fabs, while many Fabs produced hits that required optimization. Application of MMS, using the initial screens and/or refinement screens, resulted in diffraction-quality crystals of these Fabs. Five Fabs that failed to give hits in the initial screen were crystallized by cross-seeding MMS followed by MMS optimization. The crystallization protocols and strategies that resulted in structure determination of all 16 Fabs are presented. These results illustrate the power of MMS and provide a basis for developing future strategies for macromolecular crystallization.
Authors:
; ; ; ;  [1]
  1. Janssen Research and Development LLC, 1400 McKean Road, Spring House, PA 19477 (United States)
Publication Date:
OSTI Identifier:
22375697
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta crystallographica. Section F, Structural biology communications; Journal Volume: 70; Journal Issue: Pt 8; Other Information: PMCID: PMC4118815; PMID: 25084393; PUBLISHER-ID: nj5193; OAI: oai:pubmedcentral.nih.gov:4118815; Copyright (c) Obmolova et al. 2014; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CHAINS; CRYSTALLIZATION; CRYSTALS; DIFFRACTION; OPTIMIZATION; PROTEINS; SCREENING; VISIBLE RADIATION