Sinha Y N, Lewis U J
Biochem Biophys Res Commun. 1986 Oct 30;140(2):491-7. doi: 10.1016/0006-291x(86)90759-x.
By means of a lectin binding immunoassay we detected a glycosylated human growth hormone-immunoreactive substance in the human pituitary gland. The assay involves reacting the test substance with concanavalin A immobilized on a solid support followed by treatment with human growth hormone antibodies. Quantitation is achieved by reaction of the concanavalin A-glycoprotein-antibody complex with an 125I-labeled second antibody. Specificity of the assay for a glycosylated human growth hormone was indicated by inhibition of binding by N-acetyl-D-glucosamine. Furthermore, use of antiserum that had been absorbed with human growth hormone reduced the amount of radioactivity bound to the tubes. Western blotting of pituitary immunoprecipitates after gel electrophoresis in sodium dodecyl sulfate revealed human growth hormone-immunoreactive components that also reacted with concanavalin A.
通过凝集素结合免疫测定法，我们在人脑垂体中检测到一种糖基化的人生长激素免疫反应性物质。该测定法包括使测试物质与固定在固体支持物上的伴刀豆球蛋白A反应，随后用人生长激素抗体处理。通过伴刀豆球蛋白A-糖蛋白-抗体复合物与125I标记的第二抗体反应来进行定量。N-乙酰-D-葡萄糖胺对结合的抑制表明了该测定法对糖基化人生长激素的特异性。此外，使用已用人生长激素吸收的抗血清减少了与试管结合的放射性量。在十二烷基硫酸钠中进行凝胶电泳后，垂体免疫沉淀物的蛋白质印迹显示出也与伴刀豆球蛋白A反应的人生长激素免疫反应性成分。
