-
Something wrong with this record ?
Optimization of the crystallizability of a single-chain antibody fragment
J. Škerlová, V. Král, M. Fábry, J. Sedláček, V. Veverka, P. Rezáčová,
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
Free Medical Journals
from 2014 to 2 years ago
PubMed Central
from 2014 to 1 year ago
Europe PubMed Central
from 2014 to 2 years ago
- MeSH
- Chromatography, Gel MeSH
- Single-Chain Antibodies chemistry MeSH
- Crystallization * MeSH
- Humans MeSH
- Molecular Sequence Data MeSH
- Amino Acid Sequence MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
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.
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc15013966
- 003
- CZ-PrNML
- 005
- 20190130092230.0
- 007
- ta
- 008
- 150420s2014 xxu f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1107/S2053230X1402247X $2 doi
- 035 __
- $a (PubMed)25484230
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a xxu
- 100 1_
- $a Škerlová, Jana $u Institute of Molecular Genetics, ASCR, v.v.i., Vídeňská 1083, 14220 Prague 4, Czech Republic.
- 245 10
- $a Optimization of the crystallizability of a single-chain antibody fragment / $c J. Škerlová, V. Král, M. Fábry, J. Sedláček, V. Veverka, P. Rezáčová,
- 520 9_
- $a 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.
- 650 _2
- $a sekvence aminokyselin $7 D000595
- 650 _2
- $a gelová chromatografie $7 D002850
- 650 12
- $a krystalizace $7 D003460
- 650 _2
- $a lidé $7 D006801
- 650 _2
- $a molekulární sekvence - údaje $7 D008969
- 650 _2
- $a jednořetězcové protilátky $x chemie $7 D057127
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Král, Vlastimil $u Institute of Molecular Genetics, ASCR, v.v.i., Vídeňská 1083, 14220 Prague 4, Czech Republic.
- 700 1_
- $a Fábry, Milan $u Institute of Molecular Genetics, ASCR, v.v.i., Vídeňská 1083, 14220 Prague 4, Czech Republic.
- 700 1_
- $a Sedláček, Juraj $u Institute of Molecular Genetics, ASCR, v.v.i., Vídeňská 1083, 14220 Prague 4, Czech Republic.
- 700 1_
- $a Veverka, Václav $u Institute of Organic Chemistry and Biochemistry, ASCR, v.v.i., Flemingovo nám. 2, 16610 Prague 6, Czech Republic.
- 700 1_
- $a Rezáčová, Pavlína $u Institute of Molecular Genetics, ASCR, v.v.i., Vídeňská 1083, 14220 Prague 4, Czech Republic.
- 773 0_
- $w MED00189488 $t Acta crystallographica. Section F, Structural biology communications $x 2053-230X $g Roč. 70, č. Pt 12 (2014), s. 1701-6
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/25484230 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y a $z 0
- 990 __
- $a 20150420 $b ABA008
- 991 __
- $a 20190130092500 $b ABA008
- 999 __
- $a ok $b bmc $g 1071547 $s 896844
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2014 $b 70 $c Pt 12 $d 1701-6 $i 2053-230X $m Acta crystallographica. Section F, Structural biology communications $n Acta Crystallogr F Struct Biol Commun $x MED00189488
- LZP __
- $a Pubmed-20150420
