Application of region-specific immunoassay for human chromogranin A: substantial clue for detection and measurement of chromogranin A in human plasma.
Chromogranin A (CgA), a secretory protein, is co-released with catecholamines from storage vesicles. It is known to be elevated in the circulation of patients with neuroendocrine and endocrine tumors. For further investigation of the protein, especially in humans, it is essential to facilitate quantitative analysis of the protein in human biological materials. In order to introduce novel immunological methodology for this purpose, we purposely selected human CgA(344-374) for the synthetic immunogen to produce region-specific CgA antibodies. The anti-synthetic peptide antibody thus obtained made it possible to develop an immunological method for measurement and characterization of CgA in human plasma. The plasma CgA-immunoreactivity (LI) level measured by the method was 0.31+/-0.01 pmol/ml (mean+/-SEM) in normal subjects and 1.55+/-0.29 pmol/ml in pheochromocytoma. On gel chromatography and HPLC analysis of the plasma of patients with pheochromocytoma, the region-specific assay system enabled us to show the presence of N-terminal truncated CgA, besides CgA itself. By following up changes of plasma CgA-LI in a pheochromocytoma patient using samples that were collected consecutively over a two-year period, the present assay system using the region-specific antibody, anti-human CgA (344-374) serum, was confirmed to be extremely valuable for the measurement of CgA-LI in human plasma. The characteristic features and high sensitivity of the present assay system will give us a substantial clue to the detection and measurement of CgA to develop further investigation of the protein in humans.
Yanaihara
H
H
Department of Urology, Keio University School of Medicine, Tokyo, Japan.
Hata
M
M
Nishikawa
Y
Y
Hoshino
M
M
Yanaihara
N
N
Murai
M
M
D000310:Adrenal Gland Neoplasms / Q000378:metabolism / Q000628:therapy; D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000917:Antibody Formation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D053379:Chromogranin A; D002864:Chromogranins / Q000097:blood* / Q000276:immunology*; D004298:Dopamine / Q000097:blood; D004837:Epinephrine / Q000097:blood; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009638:Norepinephrine / Q000097:blood; D010673:Pheochromocytoma / Q000378:metabolism / Q000628:therapy; D011237:Predictive Value of Tests; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D004298:Dopamine; D009638:Norepinephrine; D004837:Epinephrine