Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2)
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Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). / Orskov, C; Holst, J J.
In: Scandinavian Journal of Clinical and Laboratory Investigation. Supplement, Vol. 47, No. 2, 04.1987, p. 165-74.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2)
AU - Orskov, C
AU - Holst, J J
PY - 1987/4
Y1 - 1987/4
N2 - Gene-sequencing studies have shown that the glucagon precursor contains two additional glucagon-like sequences, the so-called glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). We developed radio-immunoassays against synthetic peptides corresponding to these sequences. Antisera were raised in rabbits after carbodiimide conjugation of peptides to BSA. The selected antisera showed neither mutual cross-reactivity nor cross-reacted with any other peptide of the glucagon family. Trypsin digestion experiments showed that both antisera were directed against the C-terminus of the antigen peptides. Labelling was performed with chloramine T, and separation with plasma-coated charcoal. The detection limit was 7 and 25 pmol/l for GLP-1 and GLP-2. Accurate measurement of both peptides in plasma required extraction. Optimum recovery was obtained with ethanol at 75% (final concentration). The concentrations measured in fasting plasma from 10 normal subjects were 107 +/- 7 pmol/l and 151 +/- 14 pmol/l for GLP-1 and GLP-2, respectively. After a mixed meal the concentrations rose slowly for 2 h reaching 145 +/- 13 and 225 +/- 15 pmol/l.
AB - Gene-sequencing studies have shown that the glucagon precursor contains two additional glucagon-like sequences, the so-called glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). We developed radio-immunoassays against synthetic peptides corresponding to these sequences. Antisera were raised in rabbits after carbodiimide conjugation of peptides to BSA. The selected antisera showed neither mutual cross-reactivity nor cross-reacted with any other peptide of the glucagon family. Trypsin digestion experiments showed that both antisera were directed against the C-terminus of the antigen peptides. Labelling was performed with chloramine T, and separation with plasma-coated charcoal. The detection limit was 7 and 25 pmol/l for GLP-1 and GLP-2. Accurate measurement of both peptides in plasma required extraction. Optimum recovery was obtained with ethanol at 75% (final concentration). The concentrations measured in fasting plasma from 10 normal subjects were 107 +/- 7 pmol/l and 151 +/- 14 pmol/l for GLP-1 and GLP-2, respectively. After a mixed meal the concentrations rose slowly for 2 h reaching 145 +/- 13 and 225 +/- 15 pmol/l.
KW - Animals
KW - Chromatography, Gel
KW - Glucagon-Like Peptide 1
KW - Glucagon-Like Peptide 2
KW - Humans
KW - Hydrogen-Ion Concentration
KW - Immune Sera
KW - Peptides/blood
KW - Rabbits
KW - Radioimmunoassay/methods
M3 - Journal article
C2 - 3576119
VL - 47
SP - 165
EP - 174
JO - Scandinavian Journal of Clinical and Laboratory Investigation. Supplement
JF - Scandinavian Journal of Clinical and Laboratory Investigation. Supplement
SN - 0085-591X
IS - 2
ER -
ID: 194816936