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

Ying Wang; Yaming Shan; Xinyu Gao; Rui Gong; Jun Zheng; Xiaohua Douglas Zhang; Qi Zhao

doi:10.1016/j.molimm.2018.10.013

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

Ying Wang, Yaming Shan, Xinyu Gao, Rui Gong, Jun Zheng, Xiaohua Douglas Zhang, Qi Zhao

Biostatistics

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

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.

Original language	English
Pages (from-to)	279-285
Number of pages	7
Journal	Molecular Immunology
Volume	103
DOIs	https://doi.org/10.1016/j.molimm.2018.10.013
State	Published - Nov 2018

Bibliographical note

Publisher Copyright:
© 2018 Elsevier Ltd

Funding

This work was supported by the Science and Technology Development Fund of Macau ( FDCT/131/2016/A3 , FDCT/0015/2018/A1 ), the Guangzhou Science and Technology Program ( 201807010004 ), and Start-up Research Grand ( SRG2016-00082-FHS ) and the intramural research program of Faculty of Health Sciences, University of Macau, and National Natural Science Foundation of China ( 31440041 and 31770996 ).

Funders	Funder number
Science and Technology Development Fund of Macau	FDCT/131/2016/A3, FDCT/0015/2018/A1
National Natural Science Foundation of China (NSFC)	31770996, 31440041
Guangzhou Science and Technology Program key projects	SRG2016-00082-FHS, 201807010004
Universidade de Macau

Keywords

Antibody expression
HIV-1
Saccharomyces cerevisiae
Yeast display

ASJC Scopus subject areas

Immunology
Molecular Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.molimm.2018.10.013

Cite this

@article{3746773863a7419eb035bb32113e1811,

title = "Screening and expressing HIV-1 specific antibody fragments in Saccharomyces cerevisiae",

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",

author = "Ying Wang and Yaming Shan and Xinyu Gao and Rui Gong and Jun Zheng and Zhang, \{Xiaohua Douglas\} and Qi Zhao",

note = "Publisher Copyright: {\textcopyright} 2018 Elsevier Ltd",

year = "2018",

month = nov,

doi = "10.1016/j.molimm.2018.10.013",

language = "English",

volume = "103",

pages = "279--285",

}

TY - JOUR

T1 - Screening and expressing HIV-1 specific antibody fragments in Saccharomyces cerevisiae

AU - Wang, Ying

AU - Shan, Yaming

AU - Gao, Xinyu

AU - Gong, Rui

AU - Zheng, Jun

AU - Zhang, Xiaohua Douglas

AU - Zhao, Qi

PY - 2018/11

Y1 - 2018/11

N2 - 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.

AB - 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.

KW - Antibody expression

KW - HIV-1

KW - Saccharomyces cerevisiae

KW - Yeast display

UR - https://www.scopus.com/pages/publications/85054902469

UR - https://www.scopus.com/inward/citedby.url?scp=85054902469&partnerID=8YFLogxK

U2 - 10.1016/j.molimm.2018.10.013

DO - 10.1016/j.molimm.2018.10.013

M3 - Article

C2 - 30342371

AN - SCOPUS:85054902469

SN - 0161-5890

VL - 103

SP - 279

EP - 285

JO - Molecular Immunology

JF - Molecular Immunology

ER -

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

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this