title: Library of sequence-specific radioimmunoassays for human chromogranin A.
abstract: BACKGROUND
  Human chromogranin A (CgA) is an acidic protein widely expressed in neuroendocrine tissue and tumors. The extensive tissue- and tumor-specific cleavages of CgA at basic cleavage sites produce multiple peptides.
  
  
  METHODS
  We have developed a library of RIAs specific for different epitopes, including the NH2 and COOH termini and three sequences adjacent to dibasic sites in the remaining part of CgA.
  
  
  RESULTS
  The antisera raised against CgA(210-222) and CgA(340-348) required a free NH2 terminus for binding. All antisera displayed high titers, high indexes of heterogeneity ( approximately 1.0), and high binding affinities (Keff0 approximately 0.1 x 10(12) to 1.0 x 10(12) L/mol), implying that the RIAs were monospecific and sensitive. The concentration of CgA in different tissues varied with the assay used. Hence, in a carcinoid tumor the concentration varied from 0.5 to 34.0 nmol/g tissue depending on the specificity of the CgA assay. The lowest concentration in all tumors was measured with the assay specific for the NH2 terminus of CgA. This is consistent with the relatively low concentrations measured in plasma from carcinoid tumor patients by the N-terminal assay, whereas the assays using antisera raised against CgA(210-222) and CgA(340-348) measured increased concentrations.
  
  
  CONCLUSIONS
  Only some CgA assays appear useful for diagnosis of neuroendocrine tumors, but the entire library is valuable for studies of the expression and processing of human CgA.
authors: University Department of Clinical Biochemistry (KB3013), Rigshospitalet, Blegdamsvej 9, DK-2100 Copenhagen O, Denmark.
authors: T B
authors: TB
authors: Jensen
authors: L
authors: L
authors: Hilsted
authors: J F
authors: JF
authors: Rehfeld
mesh_terms: D000236:Adenoma / Q000097:blood / Q000737:chemistry; D000328:Adult; D000918:Antibody Specificity; D002276:Carcinoid Tumor / Q000097:blood / Q000737:chemistry; D002850:Chromatography, Gel; D053379:Chromogranin A; D002864:Chromogranins / Q000097:blood / Q000737:chemistry* / Q000276:immunology; D004064:Digestive System / Q000737:chemistry; D005260:Female; D006801:Humans; D007106:Immune Sera; D007414:Intestinal Neoplasms / Q000097:blood / Q000737:chemistry; D008297:Male; D008875:Middle Aged; D019151:Peptide Library; D010673:Pheochromocytoma / Q000097:blood / Q000737:chemistry; D010911:Pituitary Neoplasms / Q000097:blood / Q000737:chemistry; D011863:Radioimmunoassay; D012016:Reference Values; D012680:Sensitivity and Specificity
chemical_list: C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D007106:Immune Sera; D019151:Peptide Library