Ryhänen L, Rantala-Ryhänen S, Tan E M, Uitto J
Coll Relat Res. 1982 Mar;2(2):117-30. doi: 10.1016/s0174-173x(82)80028-9.
A new method for the assay of collagenase activity has been developed, whereby the collagen cleavage products, after initial collagenase digestion, are degraded further by a mixture of trypsin and alpha-chymotrypsin. The degradation products are soluble in TCA and can be conveniently separated from the remaining uncleaved collagen substrate by rapid filtration. The enzyme assay is shown to be reproducible and sensitive, and it lends itself to a convenient and rapid determination of collagenase activity in relatively large numbers of samples. The applicability of this method is demonstrated by the detection of increased collagenase activity in skin fibroblast cultures derived from a patient with recessive dystrophic epidermolysis bullosa.
已开发出一种测定胶原酶活性的新方法，即胶原酶初步消化后的胶原裂解产物，会被胰蛋白酶和α-糜蛋白酶的混合物进一步降解。降解产物可溶于三氯乙酸（TCA），通过快速过滤可方便地与剩余未裂解的胶原底物分离。该酶活性测定方法具有可重复性和敏感性，适用于相对大量样本中胶原酶活性的便捷快速测定。通过检测来自隐性营养不良性大疱性表皮松解症患者的皮肤成纤维细胞培养物中增加的胶原酶活性，证明了该方法的适用性。
