Escherichia coli surface display of single-chain antibody VRC01 against HIV-1 infection

Lin Xu Wang; Michael Mellon; Dane Bowder; Meghan Quinn; Danielle Shea; Charles Wood; Shi Hua Xiang

doi:10.1016/j.virol.2014.11.018

Escherichia coli surface display of single-chain antibody VRC01 against HIV-1 infection

Lin Xu Wang, Michael Mellon, Dane Bowder, Meghan Quinn, Danielle Shea, Charles Wood, Shi Hua Xiang

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

6 Scopus citations

Abstract

Human immunodeficiency virus type 1 (HIV-1) transmission and infection occur mainly via the mucosal surfaces. The commensal bacteria residing in these surfaces can potentially be employed as a vehicle for delivering inhibitors to prevent HIV-1 infection. In this study, we have employed a bacteria-based strategy to display a broadly neutralizing antibody VRC01, which could potentially be used to prevent HIV-1 infection. The VRC01 antibody mimics CD4-binding to gp120 and has broadly neutralization activities against HIV-1. We have designed a construct that can express the fusion peptide of the scFv-VRC01 antibody together with the autotransporter β-barrel domain of IgAP gene from Neisseria gonorrhoeae, which enabled surface display of the antibody molecule. Our results indicate that the scFv-VRC01 antibody molecule was displayed on the surface of the bacteria as demonstrated by flow cytometry and immunofluorescence microscopy. The engineered bacteria can capture HIV-1 particles via surface-binding and inhibit HIV-1 infection in cell culture.

Original language	English (US)
Pages (from-to)	179-186
Number of pages	8
Journal	Virology
Volume	475
DOIs	https://doi.org/10.1016/j.virol.2014.11.018
State	Published - Jan 5 2015

Keywords

Autotransporter
E. coli K12 UT5600
HIV-1
ScFv-VRC01
Surface display
β-Barrel domain

ASJC Scopus subject areas

Virology

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title = "Escherichia coli surface display of single-chain antibody VRC01 against HIV-1 infection",

abstract = "Human immunodeficiency virus type 1 (HIV-1) transmission and infection occur mainly via the mucosal surfaces. The commensal bacteria residing in these surfaces can potentially be employed as a vehicle for delivering inhibitors to prevent HIV-1 infection. In this study, we have employed a bacteria-based strategy to display a broadly neutralizing antibody VRC01, which could potentially be used to prevent HIV-1 infection. The VRC01 antibody mimics CD4-binding to gp120 and has broadly neutralization activities against HIV-1. We have designed a construct that can express the fusion peptide of the scFv-VRC01 antibody together with the autotransporter β-barrel domain of IgAP gene from Neisseria gonorrhoeae, which enabled surface display of the antibody molecule. Our results indicate that the scFv-VRC01 antibody molecule was displayed on the surface of the bacteria as demonstrated by flow cytometry and immunofluorescence microscopy. The engineered bacteria can capture HIV-1 particles via surface-binding and inhibit HIV-1 infection in cell culture.",

keywords = "Autotransporter, E. coli K12 UT5600, HIV-1, ScFv-VRC01, Surface display, β-Barrel domain",

author = "Wang, {Lin Xu} and Michael Mellon and Dane Bowder and Meghan Quinn and Danielle Shea and Charles Wood and Xiang, {Shi Hua}",

note = "Funding Information: This work was supported in part by the Bill and Melinda Gates Foundation to SHX. GM103509 , the Fogarty AIDS International Training and Research Program D43TW01492 and T32 AI060547 from the NIH to C.W. L.X.W. was a Fogarty Fellow. DB was an NIH Ruth Kirschstein Fellow. We would like to thank Dr. You Zhou of the Microscopy core facility for his help with the study. We would also like to thank the Nebraska Center for Virology Core facility for its support. ",

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doi = "10.1016/j.virol.2014.11.018",

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AU - Wang, Lin Xu

AU - Mellon, Michael

AU - Bowder, Dane

AU - Quinn, Meghan

AU - Shea, Danielle

AU - Wood, Charles

AU - Xiang, Shi Hua

N1 - Funding Information: This work was supported in part by the Bill and Melinda Gates Foundation to SHX. GM103509 , the Fogarty AIDS International Training and Research Program D43TW01492 and T32 AI060547 from the NIH to C.W. L.X.W. was a Fogarty Fellow. DB was an NIH Ruth Kirschstein Fellow. We would like to thank Dr. You Zhou of the Microscopy core facility for his help with the study. We would also like to thank the Nebraska Center for Virology Core facility for its support.

PY - 2015/1/5

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N2 - Human immunodeficiency virus type 1 (HIV-1) transmission and infection occur mainly via the mucosal surfaces. The commensal bacteria residing in these surfaces can potentially be employed as a vehicle for delivering inhibitors to prevent HIV-1 infection. In this study, we have employed a bacteria-based strategy to display a broadly neutralizing antibody VRC01, which could potentially be used to prevent HIV-1 infection. The VRC01 antibody mimics CD4-binding to gp120 and has broadly neutralization activities against HIV-1. We have designed a construct that can express the fusion peptide of the scFv-VRC01 antibody together with the autotransporter β-barrel domain of IgAP gene from Neisseria gonorrhoeae, which enabled surface display of the antibody molecule. Our results indicate that the scFv-VRC01 antibody molecule was displayed on the surface of the bacteria as demonstrated by flow cytometry and immunofluorescence microscopy. The engineered bacteria can capture HIV-1 particles via surface-binding and inhibit HIV-1 infection in cell culture.

AB - Human immunodeficiency virus type 1 (HIV-1) transmission and infection occur mainly via the mucosal surfaces. The commensal bacteria residing in these surfaces can potentially be employed as a vehicle for delivering inhibitors to prevent HIV-1 infection. In this study, we have employed a bacteria-based strategy to display a broadly neutralizing antibody VRC01, which could potentially be used to prevent HIV-1 infection. The VRC01 antibody mimics CD4-binding to gp120 and has broadly neutralization activities against HIV-1. We have designed a construct that can express the fusion peptide of the scFv-VRC01 antibody together with the autotransporter β-barrel domain of IgAP gene from Neisseria gonorrhoeae, which enabled surface display of the antibody molecule. Our results indicate that the scFv-VRC01 antibody molecule was displayed on the surface of the bacteria as demonstrated by flow cytometry and immunofluorescence microscopy. The engineered bacteria can capture HIV-1 particles via surface-binding and inhibit HIV-1 infection in cell culture.

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KW - β-Barrel domain

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