Rapid simultaneous determination of glucagon and insulin by capillary electrophoresis immunoassays.

A rapid capillary electrophoresis (CE) with laser-induced fluorescence (LIF) competitive immunoassay has been developed for the determination of glucagon in biological mixtures. In the assay, fluorescein-conjugated glucagon is mixed with the sample followed by addition of anti-glucagon. Free and antibody-bound, tagged glucagon could be separated in 3 s using CE to obtain quantitative determination of glucagon with a concentration detection limit of 760 pM. The assay was combined with a previously developed competitive immunoassay for insulin to produce a simultaneous immunoassay for both peptides. The method was used to determine glucagon content of islets of Langerhans.

[1]  B. Karger,et al.  Affinity probe capillary electrophoresis: analysis of recombinant human growth hormone with a fluorescent labeled antibody fragment. , 1994, Analytical chemistry.

[2]  R T Kennedy,et al.  On-line competitive immunoassay for insulin based on capillary electrophoresis with laser-induced fluorescence detection. , 1996, Analytical chemistry.

[3]  R. Kennedy,et al.  Aptamers as ligands in affinity probe capillary electrophoresis. , 1998, Analytical chemistry.

[4]  R. Kennedy,et al.  Capillary electrophoresis-based immunoassay to determine insulin content and insulin secretion from single islets of Langerhans. , 1995, Analytical chemistry.

[5]  James P. Landers,et al.  Handbook of Capillary Electrophoresis , 1993 .

[6]  J. Calvin Giddings,et al.  Unified Separation Science , 1991 .

[7]  V. Hruby,et al.  Structure-function relations in glucagon. Properties of highly purified Des-his1-, monoiodo-, and [Des-Asn28,Thr29](homoserine lactone27)-glucagon , 1975 .

[8]  D. Schmalzing,et al.  Capillary electrophoresis based immunoassays: A critical review , 1997, Electrophoresis.

[9]  R. Kennedy,et al.  On‐line competitive immunoassay based on capillary electrophoresis applied to monitoring insulin secretion from single islets of Langerhans , 1998, Electrophoresis.

[10]  D. Schmalzing,et al.  Solution-phase immunoassay for determination of cortisol in serum by capillary electrophoresis. , 1995, Clinical chemistry.

[11]  R. Kennedy,et al.  Measurement of antibody‐antigen dissociation constants using fast capillary electrophoresis with laser‐induced fluorescence detection , 1997, Electrophoresis.

[12]  R. Evangelista,et al.  Feasibility studies for simultaneous immunochemical multianalyte drug assay by capillary electrophoresis with laser-induced fluorescence. , 1994, Clinical chemistry.

[13]  B. Frank,et al.  Glucagon structure and function. I. Purification and properties of bovine glucagon and monodesmidoglucagon. , 1972, The Journal of biological chemistry.