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Optimization of the crystallizability of a single-chain antibody fragment

SYSNO ASEP	0438427
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Optimization of the crystallizability of a single-chain antibody fragment
Author(s)	Škerlová, Jana (UMG-J) Král, Vlastimil (UMG-J)_RID Fábry, Milan (UMG-J)_RID Sedláček, Juraj (UMG-J)_RID Veverka, Václav (UOCHB-X)_{RID, ORCID} Řezáčová, Pavlína (UOCHB-X)_{RID, ORCID}
Number of authors	6
Source Title	Acta Crystallographica Section F-Structural Biology and Crystallization Communications. - : Wiley - ISSN 1744-3091 Roč. 70, č. 12 (2014), s. 1701-1706
Number of pages	6 s.
Language	eng - English
Country	GB - United Kingdom
Keywords	single-chain antibody fragment ; Thermofluor assay ; differential scanning fluorimetry ; crystallizability optimization ; oligomerization ; crystallization
Subject RIV	CE - Biochemistry
R&D Projects	LK11205 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Institutional support	UOCHB-X - RVO:61388963 ; UMG-J - RVO:68378050
UT WOS	000345843300029
DOI	10.1107/S2053230X1402247X
Annotation	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.
Workplace	Institute of Organic Chemistry and Biochemistry
Contact	asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434
Year of Publishing	2015

Number of the records: 1