Abstract
Objectives
The availability of self-targeting and low immunogenic therapeutic agents is critical to efficient cancer therapy. Therefore, the development of humanized therapeutic antibodies is particularly appealing.
Results
A humanized single-chain variable fragment (scFv) antibody that can target human epidermal growth factor receptor-2 (HER2)-overexpressing cancer cells was designed and produced via expression in Pichia pastoris. The expression gave a high yield of 8 mg protein/l (with a purity of 92 %) using shake-flask cultures. Functional studies also revealed that the purified recombinant anti-HER2 scFv exhibited anti-proliferative activity and could bind efficiently to HER2-overexpressing human breast cancer cell line SKBR3.
Conclusion
The recombinant scFv offers promising therapeutic and binding efficiencies that are desirable for targeted cancer therapies.
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Acknowledgments
This work was supported by China Postdoctoral Science Foundation Grant (2013M540341), Key grant cultivating interdisciplinary studies of Chinese Education Ministry (WF1113014), National Natural Science Foundation of China (21303050 and 31471659), and Pujiang Talent Program of Shanghai Municipality (13PJD012).
Supporting information
Supplementary Table 1—Comparison of relative copy numbers of gene encoding anti-HER2 scFv in different strains using real-time PCR.
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Cao, X., Yu, H., Chen, C. et al. Expression and characterization of recombinant humanized anti-HER2 single-chain antibody in Pichia pastoris for targeted cancer therapy. Biotechnol Lett 37, 1347–1354 (2015). https://doi.org/10.1007/s10529-015-1804-6
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DOI: https://doi.org/10.1007/s10529-015-1804-6