SANDWICH IMMUNOASSAY SPECIFIC FOR THE N-TERMINAL SEQUENCE OF OSTEOCALCIN

被引：13

作者：

EGUCHI, H

HOSODA, K

KURIHARA, N

NAKAMOTO, T

KOBAYASHI, S

ARITA, M

HASEGAWA, R

OHTA, T

KIYOKI, M

YAMAJI, T

SHIRAKI, M

机构：

[1] TEIJIN LTD,BIOMED RES INST,HINO,TOKYO 191,JAPAN

[2] MEIKAI UNIV,FAC DENT,DIV PERIODONT,SAKADO,SAITAMA 35002,JAPAN

[3] RES INST & PRACTICE INVOLUT DIS,MINAMIAZUMI,NAGANO 39981,JAPAN

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1995年 / 184卷 / 02期

关键词：

SANDWICH IMMUNOASSAY; N-TERMINAL OSTEOCALCIN; SYNTHETIC PEPTIDE;

D O I：

10.1016/0022-1759(95)00094-Q

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A monoclonal antibody (10B) against the N-terminal sequence of human osteocalcin was selected to characterize its epitope and species specificity. The cross-reactivity of 10B with human and rat osteocalcin demonstrated that the reactivities of 10B with both human and rat osteocalcins were very similar. The pin technology method was used to determine the epitope and clearly demonstrated that the epitope recognized by 10B was localized to residues 12-16, the sequence of which is identical in rat and human osteocalcin molecules. This monoclonal antibody was found to be useful for designing region-specific sandwich immunoassay systems for human and rat N-terminal osteocalcin (N-OC) molecules using rabbit anti-hN20 and anti-rN20 polyclonal antibodies, respectively. The osteocalcin levels in serum determined by this N-OC method were stable during prolonged storage at 25 degrees C and the method could be usefully applied in the development of immunoassay systems for many osteocalcin molecules from many other species.

引用

页码：231 / 240

页数：10

共 18 条

[1]

Borsalino, Borzoni, Caravaggio, Rocchi, Pedrazzoni, Palummeri, Pioli, Passerie, Changes in serum osteocalcin after orthopedic surgery procedures, G. Clin. Med. Bologna, 66, (1985)

[2]

Chou, Fasman, Empirical predictions of protein conformation, Ann. Rev. Biochem., 47, (1978)

[3]

Deftos, Wolfert, Hill, Burton, Two-site assays of bone gla protein (osteolcalcin) demonstrate immunochemical deterogeneity of the intact molecule, Clin. Chem., 38, (1992)

[4]

Fujiwara, Yasuno, Kitagawa, Immunoassay for pepleomycin, a new bleomycin analog, Cancer Res., 41, (1981)

[5]

Gregory, Poser, Chemotactic activity of the γ-carboxyglutamic acid containing protein in bone, Calcified Tissue International, 35, (1983)

[6]

Hauschka, Osteocalcin and its functional domains, The Chemistry and Biology of Mineralized Tissue, (1985)

[7]

Hosoda, Eguchi, Nakamoto, Kubota, Honda, Jindai, Hasegawa, Kiyoki, Yamaji, Shiraki, Sandwich immunoassay for intact human osteocalcin, Clin. Chem., 38, (1991)

[8]

Hosoda, Okada, Hirata, Determination of antigenic epitopes recognized by four monoclonal antibodies to glutathione S transferase, Hybridoma, 9, (1989)

[9]

Houghten, General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids, Proc. Natl. Acad. Sci. USA, 82, (1985)

[10]

Huq, Tseng, Chapman, The amino acid sequence of emu osteocalcin: Gas phase sequencing of Gla-containing proteins, Biochem. Int., 15, (1987)

← 1 2 →