Radioimmunoassay of human collagenase. Specificity of the assay and quantitative determination of in vivo and in vitro human skin collagenase.
Radioimmunoassay of human collagenase. Specificity of the assay and quantitative determination of in vivo and in vitro human skin collagenase.
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人胶原酶的放射免疫测定。
DOI：
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发表时间：
1972
影响因子：
4.8
通讯作者：
J. Jeffrey
中科院分区：
文献类型：
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作者：
E. Bauer;A. Eisen;J. Jeffrey
摘要：建立了一种特异性、高灵敏度的人皮肤胶原酶放射免疫测定方法。该分析基于纯化的胶原酶或组织提取物的胶原酶与同位素标记的纯化酶的竞争，用于特定抗体结合位点，之后使用第二抗体沉淀将结合的125i标记的人皮肤胶原酶与游离酶分离。放射免疫测定法提供了2至40 ng的范围，用于测定酶的浓度，并允许在酶活性无法检测的条件下对人体皮肤粗组织提取物中的胶原酶进行定量。此外，其他人类胶原酶也可以用比酶技术高200到300倍的灵敏度进行测量。在比较研究中，各种人类和动物胶原酶与125i标记的人类皮肤胶原酶竞争抗体结合位点的能力已被评估为这些酶之间相似性的衡量标准。在免疫分析中，人类滑膜、牙龈和粒细胞胶原酶与人类皮肤胶原酶的交叉反应性是相同的，这支持了来自不同人体器官的胶原酶可能存在广泛同源性的概念。尽管来自另一种灵长类动物狒狒的胶原酶与人类酶表现出明显的交叉反应性，但来自更远亲物种的胶原酶只有在蛋白质浓度比检测人类酶所需的浓度高20到30倍时才会交叉反应。这些发现表明，当这些动物胶原酶的纯化制剂可用时，将发现与人类酶的相似之处较少。
Abstract A specific, highly sensitive radioimmunoassay has been developed for human skin collagenase. The assay is based upon competition of purified collagenase, or collagenase of tissue extracts, with isotopically labeled purified enzyme for specific antibody binding sites after which a second antibody precipitation is used to separate bound 125I-labeled human skin collagenase from free enzyme. The radioimmunoassay provides a range of 2 to 40 ng for determining enzyme concentration and has permitted quantitation of collagenase in crude tissue extracts of human skin under conditions in which enzymatic activity cannot be detected. In addition, other human collagenases can also be measured with 200 to 300 times greater sensitivity than can be achieved by enzymatic techniques. In comparative studies, the ability of a variety of human and animal collagenases to compete with 125I-labeled human skin collagenase for antibody binding sites has been assessed as a measure of the similarity between these enzymes. Human synovial, gingival, and granulocytic collagenases are identical in their cross-reactivity with human skin collagenase in the immunoassay, supporting the concept that extensive homologies probably exist among collagenases from various human organs. Although collagenase from another primate, the baboon, shows marked cross-reactivity with the human enzymes, collagenases from more distantly related species cross-react only at protein concentrations which are 20 to 30 times greater than those needed to detect the human enzymes. These findings suggest that fewer similarities to the human enzymes will be found when purified preparations of these animal collagenases become available.