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- Title
Secretory expression of a bispecific antibody targeting tumor necrosis factor and ED-B fibronectin in Pichia pastoris and its functional analysis.
- Authors
Liu, Meng-yuan; Hu, Xue-ping; Xie, Mian; Jiang, Si-jing; Li, Lu-jun; Liu, Dong-xu; Yang, Xiao-song
- Abstract
Specific targeting of tumor necrosis factor (TNF)-α antagonist to the inflamed site could increase its efficacy and reduce side-effects. Here, we constructed a bispecific diabody (BsDb) that targets TNF-α and ED-B-containing fibronectin, a fibronectin isoform specifically expressed in the pannus of the inflamed synovium in rheumatoid arthritis. BsDb was secreted from Pichia pastoris as functional protein and was purified to homogeneity. BsDb could simultaneously bind to human TNF-α and B-FN and neutralize TNF-α action. Additionally, BsDb showed a significant gain both in the antigen-binding affinity and in TNF-α-neutralizing ability as compared to its original antibodies, L19 and anti-TNF-α scFv, which were produced in E. coli. BsDb was constructed and was endowed with enhanced bioactivities and improved production processing. Therefore, it holds great potential for in vivo applications.
- Subjects
TUMOR necrosis factors; SACCHAROMYCETACEAE; ANTIGENS; PICHIA pastoris; ENDOMYCETALES
- Publication
Biotechnology Letters, 2014, Vol 36, Issue 12, p2425
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-014-1630-2