Establishment of hypersensitive radioimmunoassay for islet amyloid polypeptide using antiserum specific for its N-terminal region.
Using a synthetic N-terminal hexadecapeptide of islet amyloid polypeptide (IAPP), we prepared an antiserum specific for IAPP[1-16] and established an extremely sensitive radioimmunoassay (RIA) for the peptide with a minimum detection level of 0.26 fmol/tube. Since the N-terminal sequence of IAPP is 100% conserved in many mammalian species, the RIA is widely applicable in quantifying their IAPP. Analyses of pancreatic extracts of human and hamster using reverse-phase high performance liquid chromatography coupled with the RIA revealed that almost all pancreatic IAPP consisted of IAPP[1-37]. On the other hand, rat and mouse pancreata contained substantial amounts of IAPP[1-16] and IAPP[1-17] in addition to IAPP[1-37] as a major molecular form. In human plasma, IAPP[1-37] is the major molecular form secreted into the circulation in response to glucose administration. The RIA established in this study is promising in elucidating the physiological functions and the pathophysiological significance of IAPP in diabetes mellitus.
Shiomi
K
K
Third Department of Internal Medicine, Miyazaki Medical College, Japan.
Nakazato
M
M
Miyazato
M
M
Kangawa
K
K
Matsuo
H
H
Matsukura
S
S
D000328:Adult; D000595:Amino Acid Sequence; D000682:Amyloid / Q000032:analysis* / Q000097:blood / Q000235:genetics; D000818:Animals; D000918:Antibody Specificity; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007106:Immune Sera; D058228:Islet Amyloid Polypeptide; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010179:Pancreas / Q000737:chemistry* / Q000166:cytology; D010446:Peptide Fragments / Q000032:analysis* / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods; D051381:Rats; D012016:Reference Values; D012689:Sequence Homology, Nucleic Acid
D000682:Amyloid; D007106:Immune Sera; D058228:Islet Amyloid Polypeptide; D010446:Peptide Fragments; D015740:Calcitonin Gene-Related Peptide