Mauch H, Kümel G
Int Arch Allergy Appl Immunol. 1979;58(1):82-9. doi: 10.1159/000232176.
Despite the importance of the guinea pig as a model test animal for studying the humoral immune response in comparison to the development of cellular immunity after BCG or tuberculosis infection, no precise and sensitive method for the determination of such antibodies is available. We developed an indirect solid-phase radioimmunoassay on microtiter plates for this purpose. All assay conditions have been optimized such as selection of suitable plastic material, use of carrier substances, concentrations and amount of the antigen as well as the serum and second antibody, buffer, incubation times and all steps of the procedures for coating, washing, and measuring. The assay gives a linear relationship between antibody concentration in the serum to be tested and binding of the radiolabelled second antibody. A comparison with four conventional antibody titration methods reveals a 128- to 512-fold higher sensitivity of the RIA. Favorable aspects of this RIA as opposed to the methods hitherto used are discussed.
尽管豚鼠作为研究体液免疫反应的模型试验动物很重要,可用于对比卡介苗接种或结核感染后细胞免疫的发展情况,但目前尚无精确且灵敏的方法来测定此类抗体。为此,我们开发了一种用于微量滴定板的间接固相放射免疫测定法。所有测定条件均已优化,例如选择合适的塑料材料、使用载体物质、抗原以及血清和二抗的浓度与用量、缓冲液、孵育时间,以及包被、洗涤和测量等操作步骤。该测定法在所测血清中的抗体浓度与放射性标记二抗的结合之间呈现线性关系。与四种传统抗体滴定方法的比较表明,放射免疫测定法的灵敏度高出128至512倍。本文还讨论了这种放射免疫测定法相对于迄今所用方法的有利之处。