Li X Q, Thonar E J, Knudson W
Department of Biochemistry, Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612.
Connect Tissue Res. 1989;19(2-4):243-53. doi: 10.3109/03008208909043899.
We have developed a new ELISA to quantify hyaluronate. This ELISA takes advantage of an anti-keratan sulfate antibody to differentiate between the coated aggregating rat chondrosarcoma proteoglycan which captures the hyaluronate and the keratan sulfate-bearing aggregating proteoglycan added subsequently. Absorbance in this assay was shown to be linear to the logarithmic concentration of hyaluronate in the range of 15 to 1000 ng/ml. Pre-treatment of hyaluronate with papain or protease did not interfere with its quantification; in contrast, pre-treatment with 0.1N NaOH significantly interferes with the subsequent measurement of the hyaluronate molecules. The size of the hyaluronate molecule was found to be an important factor in quantification: only large size hyaluronate molecules could be quantified accurately. The ELISA was used to show that human lung carcinomas contain 2 to 500 times as much hyaluronate as normal lung tissue from the same patient.
我们开发了一种新的酶联免疫吸附测定法（ELISA）来定量检测透明质酸盐。这种ELISA利用抗硫酸角质素抗体来区分包被的聚集大鼠软骨肉瘤蛋白聚糖（它捕获透明质酸盐）和随后添加的含硫酸角质素的聚集蛋白聚糖。在该测定中，吸光度在15至1000 ng/ml的透明质酸盐对数浓度范围内显示为线性。用木瓜蛋白酶或蛋白酶对透明质酸盐进行预处理不会干扰其定量；相反，用0.1N氢氧化钠预处理会显著干扰随后对透明质酸分子的测量。发现透明质酸分子的大小是定量中的一个重要因素：只有大尺寸的透明质酸分子才能被准确量化。该ELISA用于表明人类肺癌中透明质酸盐的含量是同一患者正常肺组织的2至500倍。
