Wang Y.-X.,Cetral Laboratory |
Zhou L.-J.,Cetral Laboratory |
Zhang H.-R.,Cetral Laboratory |
Qu J.,Cetral Laboratory |
And 3 more authors.
Chinese Journal of Biologicals | Year: 2010
Objective: To screen human anti-bungarotoxin ScFv from large natural phage antibody library then identify. Methods: Specific antibody was screened from large natural phage antibody library by a course of "adsorption-elution- amplification" using bungarotoxin as target antigen, then determined for binding activity by ELISA, for neutralizing activity by competitive inhibition ELISA with horse antiserum against bungarotoxin, and typed by DNA fingerprint analysis and sequencing. Results: Five positive clones with binding activities to bungarotoxin were obtained after four rounds of panning, of which the highest inhibiting rate to neutralizing activity reached 8.5%. Three different DNA fingerprints were observed. Sequencing result proved that all the heavy chain variable regions of the three ScFv genes belonged to VH I, while the light chain variable regions were Vκ II, Vκ I and Vλ II. Conclusion: The anti-bungarotoxin ScFv with neutralizing activity was obtained from large natural phage antibody library.