title: Fusion Peptide Improves Stability and Bioactivity of Single Chain Antibody against Rabies Virus.
abstract: The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and halflife. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.
authors.forename: Hualong
authors.initials: H
authors.lastname: Xi
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.
authors.forename: Kaixin
authors.initials: K
authors.lastname: Zhang
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Yin
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.|Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, P.R. China.
authors.forename: Tiejun
authors.initials: T
authors.lastname: Gu
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Sun
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.
authors.forename: Linqing
authors.initials: L
authors.lastname: Shi
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.
authors.forename: Renxia
authors.initials: R
authors.lastname: Zhang
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.|Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, P.R. China.
authors.forename: Chunlai
authors.initials: C
authors.lastname: Jiang
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.|Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
authors.affiliation: National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, P.R. China.|Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, P.R. China.
authors.forename: Yongge
authors.initials: Y
authors.lastname: Wu
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000276:immunology* / Q000627:therapeutic use; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015870:Gene Expression; D006023:Glycoproteins / Q000276:immunology; D006207:Half-Life; D006801:Humans; D051379:Mice; D023421:Models, Animal; D009500:Neutralization Tests; D015202:Protein Engineering; D058849:Protein Refolding; D011818:Rabies / Q000276:immunology / Q000517:prevention & control*; D011819:Rabies Vaccines / Q000276:immunology*; D011820:Rabies virus / Q000276:immunology* / Q000472:pathogenicity; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D064166:Vaccine Potency; D014759:Viral Envelope Proteins / Q000276:immunology; D051844:alpha-Synuclein / Q000737:chemistry
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D004338:Drug Combinations; D006023:Glycoproteins; D011819:Rabies Vaccines; D011993:Recombinant Fusion Proteins; C497604:SNCA protein, human; D057127:Single-Chain Antibodies; D014759:Viral Envelope Proteins; D051844:alpha-Synuclein
keywords: Rabies; acidic tail of synuclein (ATS); mouse challenge model; rapid fluorescent focus inhibition test (RFFIT); scFv