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- Title
Protein crystallization with microseed matrix screening: application to human germline antibody Fabs.
- Authors
Obmolova, Galina; Malia, Thomas J.; Teplyakov, Alexey; Sweet, Raymond W.; Gilliland, Gary L.
- Abstract
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.
- Subjects
CRYSTALLIZATION; GERM cells; IMMUNOGLOBULINS; MACROMOLECULES; POLYMERS; X-ray diffraction
- Publication
Acta Crystallographica: Section F, Structural Biology Communications, 2014, Vol 70, Issue 8, p1107
- ISSN
2053-230X
- Publication type
Article
- DOI
10.1107/S2053230X14012552