Grønholt R, Holst J J
J Immunol Methods. 1983 Mar 11;58(1-2):83-91. doi: 10.1016/0022-1759(83)90265-x.
In some antisera raised against glucagon in rabbits a large fraction of the antibody binding sites are occupied by glucagon-like material which appears to be of endogenous origin. Six rabbits were immunized with a glucagon-albumin conjugate, boosted twice and bled 5 times during the course of 72 days. The antisera were found to contain between 110-750 nmol/l antibody-bound glucagon. The concentration of bound glucagon as well as the concentration of free antibody binding sites (titre) increased with time in 5 rabbits. By gel chromatography the bound material co-eluted with intact pancreatic glucagon (as opposed to the immunogen). Five different methods were examined for their ability to remove the ligand. Charcoal treatment of the antiserum at pH 2.5 for 48 h removed 91 +/- 5% of the bound ligand. After stripping, binding capacities (titres) and binding affinity (equilibrium constants) amounted to 63.2 +/- 19.4% and 105.0 +/- 25.4% of the respective pre-stripping values.
在一些针对兔胰高血糖素产生的抗血清中，很大一部分抗体结合位点被似乎源自内源性的胰高血糖素样物质占据。用胰高血糖素 - 白蛋白偶联物对6只兔子进行免疫，在72天的过程中进行两次加强免疫并采血5次。发现抗血清中抗体结合的胰高血糖素含量在110 - 750 nmol/l之间。5只兔子体内结合的胰高血糖素浓度以及游离抗体结合位点的浓度（效价）随时间增加。通过凝胶色谱法，结合物质与完整的胰腺胰高血糖素（与免疫原相对）共洗脱。研究了五种不同方法去除配体的能力。在pH 2.5条件下用活性炭处理抗血清48小时，可去除91±5%的结合配体。去除配体后，结合能力（效价）和结合亲和力（平衡常数）分别相当于去除配体前各自值的63.2±19.4%和105.0±25.4%。
