Production of antisera to des Asn28 Thr29 Homoser27 glucagon; The development of radioimmunoassay for total glucagon-like immunoreactivity in human plasma.

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Abstract
Antisera having a strong and strictly constant cross-reactivity for gut-GLI [glucagon-like immunoreactivity] were raised in 7 rabbits immunized with immunogen, a conjugate of bovine serum albumin and des-Asn28, Thr29, Homoser27-glucagon (CNBr-glucagon). All of the anti-CNBr-glucagon sera exhibited titer and affinity for glucagon sufficiently high enough to develop a sensitive radioimmunoassay [RIA]. The relative crossreactivity of the antisera to gut-GLI was comparable to that of antiserum K-4023 which strongly crossreacted with gut-GLI. One of the anti-CNBr-glucagon sera, OAL-196, did not react with the glucagon 19-29 fragment at all. The intra- and interassay coefficients of variation were 3.8-5.0 and 6.0-7.3%, respectively, in the RIA system for total GLI in human plasma using OAL-196. The fasting plasma total GLI was 374 .+-. 18 pg/ml. The plasma total GLI during 50 g oral glucose load in normal subjects increased significantly, whereas the plasma IRG level measured with the anti-glucagon 19-29 serum, OAL-123, assay system was lowered. In the gastrectomized subjects, plasma total GLI measured with the present assay system elicited a marked increase following an oral glucose load. The RIA using anti-CNBr-glucagon sera will be useful in measuring plasma total GLI.

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