Recombinant Expression and Functional Analysis of Nano-antibody in Lactococcus lactis Against Clostridium difficile Toxin

doi:10.13560/j.cnki.biotech.bull.1985.2018-0287

Abstract

Abstract: This study is to construct recombinant Lactococcus lactis expressing nanobody against Clostridium difficile toxin. The nano-antibody is a kind of natural antibody（named E3 and AA6 respectively）which is deficient in light chain and isolated from peripheral blood after immunizing alpaca. The recombinant plasmid pNZ8148-E3-AA6 was constructed and transformed into L. lactis NZ9000 to induce expression. About 32 kD target protein was detected by SDS-PAGE and Western blot. The recombinant protein E3-AA6 was purified by nickel-column affinity chromatography and its concentration was measured by BCA protein assay kit. The cell rounding experiment and CCK8 experiment were used to identify its function,E3-AA6 effectively inhibited the C. difficile toxin TcdB. In conclusion,the pNZ8148-E3-AA6 is successfully constructed and bivalent antibodies E3-AA6 is efficiently expressed in the NICE system,this provides a new therapeutic approach for the treatment of infections by C. difficile.

Key words: Lactococcus lactis, nano antibody, Clostridium difficile, NICE expression

WU Guo-song, LI Shan. Recombinant Expression and Functional Analysis of Nano-antibody in Lactococcus lactis Against Clostridium difficile Toxin[J]. Biotechnology Bulletin, 2018, 34(10): 87-91.

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