Optimization of the crystallizability of a single-chain antibody fragment

0438427 - ÚOCHB 2015 RIV GB eng J - Journal Article
Škerlová, Jana - Král, Vlastimil - Fábry, Milan - Sedláček, Juraj - Veverka, Václav - Řezáčová, Pavlína
Optimization of the crystallizability of a single-chain antibody fragment.
Acta Crystallographica Section F-Structural Biology and Crystallization Communications. Roč. 70, č. 12 (2014), s. 1701-1706. ISSN 1744-3091. E-ISSN 2053-230X
R&D Projects: GA MŠMT(CZ) LK11205
Institutional support: RVO:61388963 ; RVO:68378050
Keywords : single-chain antibody fragment * Thermofluor assay * differential scanning fluorimetry * crystallizability optimization * oligomerization * crystallization
Subject RIV: CE - Biochemistry
Impact factor: 0.524, year: 2014

Single-chain variable antibody fragments (scFvs) are molecules with immense therapeutic and diagnostic potential. Knowledge of their three-dimensional structure is important for understanding their antigen-binding mode as well as for protein-engineering approaches such as antibody humanization. A major obstacle to the crystallization of single-chain variable antibody fragments is their relatively poor homogeneity caused by spontaneous oligomerization. A new approach to optimization of the crystallizability of single-chain variable antibody fragments is demonstrated using a representative single-chain variable fragment derived from the anti-CD3 antibody MEM-57. A Thermofluor-based assay was utilized to screen for optimal conditions for antibody-fragment stability and homogeneity. Such an optimization of the protein storage buffer led to a significantly improved ability of the scFv MEM-57 to yield crystals.
Permanent Link: http://hdl.handle.net/11104/0242439