PROPERTIES OF A SINGLE-CHAIN ANTIBODY CONTAINING DIFFERENT LINKER PEPTIDES

被引:69
作者
ALFTHAN, K
TAKKINEN, K
SIZMANN, D
SODERLUND, H
TERRI, TT
机构
[1] VTT Biotechnology and Food Research, FIN-02044 VTT, Espoo
[2] Hoffmann-La Roche AG, Betriebe Kaiseraugst, Bau 205/211
来源
PROTEIN ENGINEERING | 1995年 / 8卷 / 07期
关键词
ANTIBODY; LINKER DESIGN; MULTIMERIZATION; PROTEOLYTIC STABILITY; SINGLE-CHAIN SCFV;
D O I
10.1093/protein/8.7.725
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Single-chain antibodies were constructed using six different linker peptides to join the V-H and V-L domains of an anti-2-phenyloxazolone (Ox) antibody, Four of the linker peptides originated from the interdomain linker region of the fungal cellulase CBHI and consisted of 28, 11, six and two amino acid residues, The two other linker peptides used were the (GGGGS)(3) linker with 15 amino acid residues and a modified IgG2b hinge peptide with 22 residues, Proteolytic stability and Ox binding properties of the six different scFv derivatives produced in Escherichia coli were investigated and compared with those of the corresponding Fv fragment containing no joining peptide between the V domains, The hapten binding properties of different antibody fragments were studied by ELISA and BIAcore(TM). The interdomain linker peptide improved the hapten binding properties of the antibody fragment when compared with Fv fragment, but slightly increased its susceptibility to proteases, Single-chain antibodies with short CBHI linkers of 11, six and two residues had a tendency to form multimers which led to a higher apparent affinity, The fragments with linkers longer than 11 residues remained monomeric.
引用
收藏
页码:725 / 731
页数:7
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