Article

A novel single-chain Fv antibody for connective tissue growth factor against the differentiation of fibroblast into myofibroblast.

Center of Clinical Laboratory Medicine of Zhongda Hospital, Southeast University, Nanjing, China.
Applied Microbiology and Biotechnology (impact factor: 3.42). 12/2011; 93(6):2475-82. DOI:10.1007/s00253-011-3755-8 pp.2475-82
Source: PubMed

ABSTRACT This study was aimed to investigate the effect of a single-chain fragment variable antibody of connective tissue growth factor (anti-CTGF scFv) against the differentiation of fibroblast into myofibroblast. The scFv antibody was firstly expressed in Escherichia coli cells and was then purified by affinity chromatography. The yield scFv protein reached a purity over 95% after purification. Immunoreactivity assay demonstrated that scFv possessed a special affinity toward CTGF. RT-PCR, western blot, and immunofluorescence experiments showed that increased expression of α-smooth muscle actin induced by TGF-β1 could be suppressed by this scFv antibody through inhibiting the phosphorylation of Akt.

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Keywords

affinity chromatography
 
Akt
 
anti-CTGF scFv
 
connective tissue growth factor
 
CTGF
 
immunofluorescence experiments
 
Immunoreactivity assay
 
purity
 
scFv antibody
 
single-chain fragment variable antibody
 
special affinity
 
western blot
 
α-smooth muscle actin induced
 

Guoqiu Wu