Post by Creative Diagnostics

Recombinant Antibodies with Unique Specificities for the Detection of Uncarboxylated Osteocalcin - Creative Diagnostics We have developed recombinant antibodies (scFv and Fab) with carefully characterized epitopes on human uncarboxylated osteocalcin • scFvs were screened from a phage display library by positive selection for uncarboxylated osteocalcin (ucOC) and negative selection for carboxylated osteocalcin (cOC) • scFvs (and later Fabs) were cloned into expression vector as a fusion protein with bacterial alkaline phosphatase (AP), His(6)tag and Flag-tag • Fab-APs were produced in BL21 E. coli (IPTG induction for 10 hrs) and purified using Ni-NTA columns • 4 clones were produced and characterized in detail (clones 2, 13, 16, 19)