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Mol Immunol. 2018 Nov;103:279-285. doi: 10.1016/j.molimm.2018.10.013. Epub 2018 Oct 17.

Screening and expressing HIV-1 specific antibody fragments in Saccharomyces cerevisiae.

Author information

1
Institute of Chinese Medical Sciences, University of Macau, Macau, China.
2
National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China.
3
Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
4
Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
5
Cancer Centre, Faculty of Health Sciences, University of Macau, Macau, China; Institute of Translational Medicine, Faculty of Heath Sciences, University of Macau, Macau, China.
6
Cancer Centre, Faculty of Health Sciences, University of Macau, Macau, China; Institute of Translational Medicine, Faculty of Heath Sciences, University of Macau, Macau, China. Electronic address: zhaoqi@alumni.cuhk.net.

Abstract

Yeast displaying techniques have been widely used for identifying novel single-chain variable fragments (scFvs) and engineering their binding properties. In this study, we establish a set of vectors for scFv screening and production in the yeast system of Saccharomyces cerevisiae. This suite includes a display vector pYS for screening of recombinant scFv libraries as well as an expression vector pYE for production of scFv candidates in Saccharomyces cerevisiae. The display vector, pYS, give the identification of the HIV-1-specific scFv clones from one scFv display library by fluorescence-activated cell sorting. Subsequently, the expression vector pYE can offer high quality scFvs of interest up to hundreds of microgram scale for bioactivity analysis. As the result, one identified scFv was confirmed to exhibit HIV-1 neutralization activity in a cell line-based pseudovirus assay. The advantage of this system enables the identical post-translation of mammalian scFvs in the same host cells. Therefore, this vector set can be useful for the rapid screening and expression of antibody genes.

KEYWORDS:

Antibody expression; HIV-1; Saccharomyces cerevisiae; Yeast display

PMID:
30342371
DOI:
10.1016/j.molimm.2018.10.013
[Indexed for MEDLINE]

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