title: Production of monoclonal and polyclonal antibodies against human osteocalcin sequences and development of a two-site ELISA for intact human osteocalcin.
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.
authors.affiliation: Department of Biochemistry and Biotechnology, University of Kuopio, Finland.
authors.forename: I
authors.initials: I
authors.lastname: Kuronen
authors.forename: H
authors.initials: H
authors.lastname: Kokko
authors.forename: M
authors.initials: M
authors.lastname: Parviainen
mesh_terms: D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D002645:Chickens / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D006825:Hybridomas; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D007136:Immunoglobulins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D008297:Male; D051379:Mice / Q000276:immunology; D008875:Middle Aged; D015675:Osteocalcin / Q000097:blood* / Q000276:immunology*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine
chemical_list: D000911:Antibodies, Monoclonal; C016512:IgY; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007136:Immunoglobulins; D015675:Osteocalcin; D012710:Serum Albumin, Bovine