Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice

Abstract

Butyrylcholinesterase (BCHE) is a natural bioscavenger that protects humans against organophosphate toxicity. Due to the limited yield of human BCHE (hBCHE) when purifying from human plasma, it is necessary to find an alternative method to produce this protein. One potential method is to produce transgenic livestock that make modified milk containing high concentration of hBCHE. In this study, we cloned the hBCHEgene into a human lactoferrin (hLF) bacterial artificial chromosome (BAC) construct to make a hLF-hBCHE BAC construct. Subsequently, we injected the BAC construct into pronuclei of mouse fertilized embryos and generated transgenic mice. Expression analysis showed that recombinant hBCHE (rhBCHE) was expressed efficiently in the mammary gland of the transgenic mice and the concentration of rhBCHE in the milk of individual mice ranged from 76±12 to 159±28 mg·L^-1. Protein function tests showed that rhBCHE has the same enzymatic activity as the native hBCHE. Our results pave the way for making transgenic livestock to produce large quantities of rhBCHE.

Keywords

recombinant human butyrylcholinesterase (rhBCHE) / human lactoferrin bacterial artificial chromosome (hLF BAC) / transgenic mice / milk

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Acknowledgements

We wish to thank Min Zheng and Meili Wang for their excellent technical assistance and all of our colleagues for their valuable comments and suggestions.

Supplementary material

The online version of this article at http://dx.doi.org/(doi:10.15302/J-FASE-2014020) contains supplementary material (Appendix A).

Compliance with ethics guidelines

Dan Lu, Shengzhe Shang, Shen Liu, Ying Wu, Fangfang Wu, Tan Tan, Qiuyan Li, Yunping Dai, Xiaoxiang Hu, Yaofeng Zhao and Ning Li declare that they have no conflict of interest or financial conflicts to disclose.

All applicable institutional and national guidelines for the care and use of animals were followed.

RIGHTS & PERMISSIONS

Higher Education Press and Springer-Verlag Berlin Heidelberg