Claassen E, Van Rooijen N
J Histochem Cytochem. 1985 Aug;33(8):840-4. doi: 10.1177/33.8.3894501.
A new conjugate for the detection of anti-trinitrophenyl(TNP) antibodies was developed to study the localization pattern of specific antibody containing cells and extracellular antibody in vivo. By means of a bridging molecule, poly-L-lysine, nine TNP groups and six horseradish peroxidase (HRP) groups were joined in one conjugate. Thus a higher specificity (more hapten) was united with a higher staining intensity (more enzyme) in the same conjugate. This conjugate made possible the simultaneous detection of anti-TNP antibody containing cells and establishment of their class (immunoglobulin M (IgM) and IgG). It was also used for the demonstration of anti-TNP antibodies in tissues where a TNP-alkaline phosphate (AP) conjugate could not be used due to high AP (endogenous) background staining. Thus we demonstrated anti-TNP antibody containing cells in gut associated lymphoid tissue and anti-TNP-(TNP-ovalbumin) immune complexes in the glomeruli of the kidney. We suggest that poly-L-lysine is a suitable bridging molecule for the preparation of hapten-HRP conjugates.
为了研究体内含特异性抗体细胞和细胞外抗体的定位模式，开发了一种用于检测抗三硝基苯基（TNP）抗体的新型偶联物。借助桥接分子聚-L-赖氨酸，将九个TNP基团和六个辣根过氧化物酶（HRP）基团连接在一个偶联物中。因此，在同一偶联物中，更高的特异性（更多半抗原）与更高的染色强度（更多酶）相结合。这种偶联物使得同时检测含抗TNP抗体的细胞并确定其类别（免疫球蛋白M（IgM）和IgG）成为可能。它还用于在由于高碱性磷酸酶（内源性）背景染色而无法使用TNP-碱性磷酸酶（AP）偶联物的组织中显示抗TNP抗体。因此，我们在肠道相关淋巴组织中证实了含抗TNP抗体的细胞，并在肾小体中证实了抗TNP-（TNP-卵清蛋白）免疫复合物。我们认为聚-L-赖氨酸是制备半抗原-HRP偶联物的合适桥接分子。
