A new human chromogranin A (CgA) immunoradiometric assay involving monoclonal antibodies raised against the unprocessed central domain (145-245).
Chromogranin A (CgA), a major protein of chromaffin granules, has been described as a potential marker for neuroendocrine tumours. Because of an extensive proteolysis which leads to a large heterogeneity of circulating fragments, its presence in blood has been assessed in most cases either by competitive immunoassays or with polyclonal antibodies. In the present study, 24 monoclonal antibodies were raised against native or recombinant human CgA. Their mapping with proteolytic peptides showed that they defined eight distinct epitopic groups which spanned two-thirds of the C-terminal part of human CgA. All monoclonal antibodies were tested by pair and compared with a reference radioimmunoassay (RIA) involving CGS06, one of the monoclonal antibodies against the 198-245 sequence. It appears that CgA C-terminal end seems to be highly affected by proteolysis and the association of C-terminal and median-part monoclonal antibodies is inadequate for total CgA assessment. Our new immunoradiometric assay involves two monoclonal antibodies, whose contiguous epitopes lie within the median 145-245 sequence. This assay allows a sensitive detection of total human CgA and correlates well with RIA because dibasic cleavage sites present in the central domain do not seem to be affected by degradation. It has been proved to be efficient in measuring CgA levels in patients with neuroendocrine tumours.
CIS Biointernational, Division In Vitro Technologies, Bagnols-sur-Cèze, France.
F
F
Degorce
Y
Y
Goumon
L
L
Jacquemart
C
C
Vidaud
L
L
Bellanger
D
D
Pons-Anicet
P
P
Seguin
M H
MH
Metz-Boutigue
D
D
Aunis
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D053379:Chromogranin A; D002864:Chromogranins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D015592:Immunoradiometric Assay / Q000379:methods*; D008666:Metalloendopeptidases / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018358:Neuroendocrine Tumors / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D000911:Antibodies, Monoclonal; C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C506155:chromogranin A, mouse; D008666:Metalloendopeptidases; C039830:peptidyl-Lys metalloendopeptidase