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.
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
authors.forename: Lin-Xu
authors.initials: LX
authors.lastname: Wang
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Veterinary Medicine and Biomedical Sciences, Lincoln, NE, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Mellon
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
authors.forename: Dane
authors.initials: D
authors.lastname: Bowder
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Veterinary Medicine and Biomedical Sciences, Lincoln, NE, USA.
authors.forename: Meghan
authors.initials: M
authors.lastname: Quinn
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Shea
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Wood
authors.affiliation: Nebraska Center for Virology, Lincoln, NE, USA; School of Veterinary Medicine and Biomedical Sciences, Lincoln, NE, USA. Electronic address: sxiang2@unl.edu.
authors.forename: Shi-Hua
authors.initials: SH
authors.lastname: Xiang
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism*; D057134:Antibodies, Neutralizing / Q000502:physiology; D001665:Binding Sites; D000080908:Broadly Neutralizing Antibodies; D015704:CD4 Antigens; D004926:Escherichia coli / Q000378:metabolism*; D015483:HIV Antibodies / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000378:metabolism; D015658:HIV Infections / Q000276:immunology / Q000517:prevention & control; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D008958:Models, Molecular; D009344:Neisseria gonorrhoeae; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000080908:Broadly Neutralizing Antibodies; D015704:CD4 Antigens; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C578595:VRC01 monoclonal antibody; C514793:gp120 protein, Human immunodeficiency virus 1; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase
keywords: Autotransporter; E. coli K12 UT5600; HIV-1; Surface display; scFv-VRC01; β-Barrel domain