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Secretory expression of a bispecific antibody targeting tumor necrosis factor and ED-B fibronectin in Pichia pastoris and its functional analysis

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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.

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Acknowledgments

This work was supported by grant of National Nature Science Foundation of China (30973669/H3004).

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Authors and Affiliations

  1. Center for Infection and Immunity Research, School of Life Sciences, Hubei University, Youyi Road 368, Wuhan, 430062, China

    Meng-yuan Liu, Xue-ping Hu, Mian Xie, Si-jing Jiang, Lu-jun Li, Dong-xu Liu & Xiao-song Yang

  2. Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Youyi Road 368, Wuhan, 430062, China

    Meng-yuan Liu, Xue-ping Hu, Mian Xie, Si-jing Jiang, Lu-jun Li, Dong-xu Liu & Xiao-song Yang

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  1. Meng-yuan Liu
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  2. Xue-ping Hu
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  3. Mian Xie
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  4. Si-jing Jiang
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  5. Lu-jun Li
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  6. Dong-xu Liu
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  7. Xiao-song Yang
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Correspondence to Meng-yuan Liu.

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Liu, My., Hu, Xp., Xie, M. et al. Secretory expression of a bispecific antibody targeting tumor necrosis factor and ED-B fibronectin in Pichia pastoris and its functional analysis. Biotechnol Lett 36, 2425–2431 (2014). https://doi.org/10.1007/s10529-014-1630-2

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  • DOI: https://doi.org/10.1007/s10529-014-1630-2

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