title: Engineering of a novel zipFv using leucine zipper motif against rabies virus glycoprotein G with improved protection potency in vivo.
abstract: Rabies is an acute zoonotic infectious disease with a high fatality rate but is preventable with vaccination and rabies immunoglobulin (RIG). The single-chain Fv fragment (scFv), a small engineered antigen-binding protein derived from antibody variable heavy (VH) and light (VL) chains connected by a peptide linker, can potentially be used to replace RIG. Here, we produced two peptides VH-JUN-HIS and VL-FOS-HA separately in Escherichia coli and assembled them to form zipFv successfully in vitro. The new zipFv utilizes FOS and JUN leucine zippers to form an antibody structure similar to the IgG counterpart with two free N-terminal ends of VHand VL. The zipFv protein showed notable improvement in binding ability and affinity over its corresponding scFv. The zipFv also demonstrated greater stability in serum and the same protective rate as RIG against challenge with a standard rabies virus (CVS-24) in mice. Our results indicated zipFv as a novel and efficient antibody form with enhanced neutralizing potency.
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China.
authors.forename: Hualong
authors.initials: H
authors.lastname: Xi
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China.
authors.forename: Kaixin
authors.initials: K
authors.lastname: Zhang
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China.
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Yin
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China.
authors.forename: Tiejun
authors.initials: T
authors.lastname: Gu
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Sun
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China.
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Li
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China.
authors.forename: Chunlai
authors.initials: C
authors.lastname: Jiang
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China. Electronic address: ygwu@jlu.edu.cn.
authors.forename: Yongge
authors.initials: Y
authors.lastname: Wu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics; D000915:Antibody Affinity; D000956:Antigens, Viral / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D016350:Leucine Zippers / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D015202:Protein Engineering; D055550:Protein Stability; D011818:Rabies / Q000276:immunology*; D011819:Rabies Vaccines / Q000235:genetics / Q000276:immunology*; D011820:Rabies virus / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000235:genetics; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D011819:Rabies Vaccines; D057127:Single-Chain Antibodies; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus
keywords: Leucine zipper; RFFIT; Rabies; scFv; zipFv