[Screening of single-chain variable fragment (scFv) to bone sialoprotein].

AIM

To select single-chain variable fragment(scFv) antibody specific for bone sialoprotein(BSP) from Human Single Fold scFv Libraries.

METHODS

Human Single Fold scFv Libraries were panned against immobilized BSP in a microtiter plate, after three rounds of panning, 96 clones were determined specific to BSP. The specificity of each scFv clone was determined by ELISA. The coding gene for BSP protein scFv has been sequenced.

RESULTS

Phage antibody for BSP protein had a specific combination character. There were 368 bp, 527 bp, 935 bp which werer light chain, heavy chain and joint gene fragment with the resuLt of PCR. The DNA sequence data showed that there were 11 differences of the amino acids in the light chain, while there were only 3 differences in the heavy chain of scFv.

CONCLUSION

scFv specific to BSP has been identified by means of phage display technology.