Dusemund B, Barrach H J
J Immunol Methods. 1982;50(3):255-68. doi: 10.1016/0022-1759(82)90163-6.
Specific quantification of collagen types I and II by sandwich ELISA or double-sandwich ELISA is described. Specific anticollagen antibodies are linked to polystyrene microplates. The collagen to be measured binds to the coating antibodies. Collagen type-specific second antibodies react with the immobilized antigen. The second antibodies are either labeled with peroxidase or are detected by anti-IgG antibodies conjugated with peroxidase. Bound peroxidase is estimated by the color reaction produced with the substrate 5-aminosalicylic acid. Optimization of the test procedure was achieved by varying the conditions for coating, antigen and second antibody incubations. Sensitivity, precision, specificity and recovery of the method have been analyzed. The assay is compared with inhibition ELISA and hydroxyproline determination.
本文描述了通过夹心ELISA或双夹心ELISA对I型和II型胶原蛋白进行特异性定量的方法。特异性抗胶原蛋白抗体与聚苯乙烯微孔板相连。待检测的胶原蛋白与包被抗体结合。胶原蛋白类型特异性的二抗与固定化抗原反应。二抗要么用辣根过氧化物酶标记，要么用与辣根过氧化物酶偶联的抗IgG抗体进行检测。通过底物5-氨基水杨酸产生的颜色反应来估算结合的辣根过氧化物酶。通过改变包被、抗原和二抗孵育的条件实现了检测程序的优化。分析了该方法的灵敏度、精密度、特异性和回收率。将该检测方法与抑制ELISA和羟脯氨酸测定法进行了比较。
