Isolation of glucagon-like immunoreactivity of gut by affinity chromatography on anti-glucagon antibodies coupled to sepharose 4B

Secretin and insulin: Response to intraduodenal acid

“…Two antibodies were used in the studies BB33 raised against synthetic secretin (final titre 1:20,000 to 36,000); and BB101 raised against natural secretin (final titre 1 : 36,000). The antibodies appeared specific for secretin in that no cross-reaction was noted with pancreatic glucagon, large gut glucagon-like immunoreactivity [11] (GLI), human insulin (Medical Research Council), gastric inhibitory polypeptide and motilin (both kindly donated by Dr. J. C. Brown), 90 per cent pure cholecystokinin-pancreozymin (kindly donated by Dr. V. Mutt), human synthetic gastrin (Imperial Chemical Industries) and 9 ~asoactive intestinal polypeptide (kindly donated by Dr. V. Mutt).…”

Insulin releasing activity of gastrointestinal glucagon-like immunoreactive materials in perfused rat pancreas.

“…Although the biological activity of GLI in the gastrointestinal tract was still a matter of speculation, controversy continued as to whether the intestinal GLI possesses the insulinogenic activity (Buchanan et al, 1969;Moody et al, 1970;Gutman et al, 1973;Murphy et al, 1973;Tanaka et al, 1975b;Ohneda et al, 1976). Conflicting results regarding this matter were partly due to a difference in the purity of the material employed and the system to study.…”

Glucagon levels in normal and diabetic subjects: Use of a specific immunoabsorbent for glucagon radioimmunoassay

“…The antisera produced reacted both with pancreatic glucagon and enteroglucagon (synonymous with gut glucagon and glucagon-like immunoreactivity of intestinal origin, or GLI) as indicated by full displacement of labelled pancreatic glucagon from the antibody with both pancreatic glucagon standards and enteroglucagon containing intestinal extracts [13]. The antisera did not cross-react with other gut hormones.…”

“…The antisera did not cross-react with other gut hormones. A globulin concentrate was prepared by ammonium sulphate precipitation, redissolved in 1 ml of 0.1 M bicarbonate -0.14 M NaC1, pH 8.0 buffer and, after extensive dialysis, coupled to cyanogen bromide activated sepharose 4B beads (Pharmacia, Uppsala, Sweden) [13]. The globulin (80 rag) was coupled to 20 ml (wet volume) activated sepharose beads by mixing for 15 h at 4 ~ C. The antibody complexed sepharose was then washed alternatively with the bicarbonate-saline buffer (2L) and distilled water (2L) and unreacted cyanogen bromide groups were blocked by 2 h incubation with 200 ml 1M ethanolamine (pH 8.0).…”