title: Production and characterization of a recombinant single-chain antibody against Hantaan virus envelop glycoprotein.
abstract: Hantaan virus (HTNV) is the type of Hantavirus causing hemorrhagic fever with renal syndrome, for which no specific therapeutics are available so far. Cell type-specific internalizing antibodies can be used to deliver therapeutics intracellularly to target cell and thus, have potential application in anti-HTNV infection. To achieve intracellular delivery of therapeutics, it is necessary to obtain antibodies that demonstrate sufficient cell type-specific binding, internalizing, and desired cellular trafficking. Here, we describe the prokaryotic expression, affinity purification, and functional testing of a single-chain Fv antibody fragment (scFv) against HTNV envelop glycoprotein (GP), an HTNV-specific antigen normally located on the membranes of HTNV-infected cells. This HTNV GP-targeting antibody, scFv3G1, was produced in the cytoplasm of Escherichia coli cells as a soluble protein and was purified by immobilized metal affinity chromatography. The purified scFv possessed a high specific antigen-binding activity to HTNV GP and HTNV-infected Vero E6 cells and could be internalized into HTNV-infected cells probably through the clathrin-dependent endocytosis pathways similar to that observed with transferrin. Our results showed that the E. coli-produced scFv had potential applications in targeted and intracellular delivery of therapeutics against HTNV infections.
authors.affiliation: Department of Nephrology, Tangdu Hospital, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.forename: Junxia
authors.initials: J
authors.lastname: Wei
authors.forename: Fanglin
authors.initials: F
authors.lastname: Zhang
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Lintao
authors.initials: L
authors.lastname: Jia
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.forename: Wen
authors.initials: W
authors.lastname: Luo
authors.forename: Yunxin
authors.initials: Y
authors.lastname: Cao
authors.forename: Libo
authors.initials: L
authors.lastname: Yao
authors.forename: Jifeng
authors.initials: J
authors.lastname: Sun
authors.forename: Zhikai
authors.initials: Z
authors.lastname: Xu
authors.forename: Angang
authors.initials: A
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D004705:Endocytosis; D004926:Escherichia coli / Q000235:genetics; D018063:Hantaan virus / Q000276:immunology*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014759:Viral Envelope Proteins