Lindenhayn K, Haupt R, Heilmann H H, Wolf H
Biomed Biochim Acta. 1987;46(1):33-8.
The radial diffusion assay is a very useful method for detection of low amounts of proteinase inhibitors in biological materials. The determination of low molecular weight (LMW) inhibitors in the presence of high molecular weight inhibitors is possible by the combination of radial diffusion and ultrafiltration. Using this method LMW trypsin inhibitors could be demonstrated in human articular cartilage, but not in human synovial fluid. In cow, pig and sheep a LMW trypsin inhibitor could be found in both the articular cartilage and in the synovial fluid. On the other hand, a LMW trypsin inhibitor could not be found neither in the canine cartilage nor in the canine synovial fluid. The method allows also the direct determination of LMW trypsin inhibitors in cartilage extracts in the presence of 4 M guanidinium hydrochloride or 6 M urea. Therefore, the method is recommended for direct determination of LMW inhibitors by column chromatographic separations of inhibitors.
径向扩散测定法是检测生物材料中低含量蛋白酶抑制剂的一种非常有用的方法。通过径向扩散和超滤相结合，可以在存在高分子量抑制剂的情况下测定低分子量（LMW）抑制剂。使用这种方法，在人关节软骨中可检测到LMW胰蛋白酶抑制剂，但在人滑液中未检测到。在牛、猪和羊的关节软骨和滑液中均能发现LMW胰蛋白酶抑制剂。另一方面，在犬类软骨和犬类滑液中均未发现LMW胰蛋白酶抑制剂。该方法还允许在存在4M盐酸胍或6M尿素的情况下直接测定软骨提取物中的LMW胰蛋白酶抑制剂。因此，推荐该方法用于通过抑制剂的柱色谱分离直接测定LMW抑制剂。
