Kuronen I, Kokko H, Parviainen M
J Immunol Methods 1993 Aug 163:233-40
Abstract
In this study we have raised polyclonal and monoclonal antibodies against human osteocalcin sequences coupled to bovine serum albumin (BSA). The antibodies were used for the development of a novel two-site ELISA on microtiter plates for intact human osteocalcin. The epitope, recognized by the monoclonal hybridoma product Os 31/2 that was used as the capture antibody, was located on the amino terminal sequence 1-29. The antibodies used for detection recognized the sequence 1-49. This method failed to detect any tryptic peptides derived from human osteocalcin sequence 1-49 nor did it detect peptide 1-29, thus demonstrating high specificity for the intact osteocalcin 1-49 only. This is a novel technique for the determination of intact osteocalcin. We have also validated the assay parameters for routine use. This direct and relatively simple procedure promises to be a powerful diagnostic and investigative tool. | 
