Single-chain Fv antibody fragments retain binding properties of the monoclonal antibody raised against peptide P1 of the human prion protein.
Prion diseases are incurable neurodegenerative diseases that affect both humans and animals. The infectious agent is a pathogenic form of the prion protein that accumulates in brain as amyloids. Currently, there is neither cure nor reliable preclinical diagnostics on the market available. The growing number of reports shows that passive immunisation is one of the most promising strategies for prion disease therapy, where antibodies against prions may prevent and even cure the infection. Since antibodies are large molecules and, thus, might not be suitable for the therapy, different antibody fragments are a good alternative. Therefore, we have designed and prepared single-chain antibody fragments (scFvs) derived from the PrP(Sc)-specific murine monoclonal antibody V5B2. Using a new expression vector pMD204, we produced scFvs in two opposing chain orientations in the periplasm of Escherichia coli. Both recombinant antibody fragments retained the specificity of the parent antibody and one of these exhibited binding properties comparable to the corresponding murine Fab fragments with the affinity in nM range. Our monovalent antibody fragments are of special interest in view of possible therapeutic reagents for prion diseases as well as for development of a new generation of diagnostics.
Skrlj
Nives
N
Chair of Biochemistry, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Jamova 39, 1000, Ljubljana, Slovenia.
Serbec
Vladka Curin
VC
Dolinar
Marko
M
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D011328:Prions / Q000276:immunology*; D055550:Protein Stability; D044967:Serum; D057127:Single-Chain Antibodies / Q000276:immunology* / Q000302:isolation & purification; D013696:Temperature
D000911:Antibodies, Monoclonal; D000941:Antigens; D007135:Immunoglobulin Variable Region; D010455:Peptides; D011328:Prions; D057127:Single-Chain Antibodies