Yoneda Kazuhito, Nakano Masakazu, Mori Kazuhiko, Kinoshita Shigeru, Tashiro Kei
Department of Genomic Medical Sciences, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Growth Factors. 2007 Jun;25(3):160-7. doi: 10.1080/08977190701723505.
TGF-beta3 has been implicated in the pathology of ocular diseases, but its concentration in human aqueous humor has never been assessed. In this study, we established an enzyme-linked immunosorbent assay (ELISA) for TGF-beta3 and quantitated it in aqueous humor collected from patients with pseudoexfoliation syndrome (PE), primary open angle glaucoma, chronic angle closure glaucoma and cataract (as the control). To develop the TGF-beta3 ELISA, we screened antibodies to identify the best combination, validated the assay for aqueous humor, and optimized the procedure for preparing activated TGF-beta3. As a result, our ELISA was highly selective and reproducible. Using our ELISA, we discovered a significantly elevated concentration of TGF-beta3 in PE eyes. We also developed new TGF-beta1 and -beta2 ELISAs, and were able, for the first time, to quantitate all the TGF-beta isoforms in the aqueous humor from a single eye, to assess their proportional effects on the pathogenesis of ocular diseases.
转化生长因子β3（TGF-β3）与眼部疾病的病理过程有关，但其在人房水中的浓度从未被评估过。在本研究中，我们建立了一种用于检测TGF-β3的酶联免疫吸附测定法（ELISA），并对从假性剥脱综合征（PE）、原发性开角型青光眼、慢性闭角型青光眼患者以及白内障患者（作为对照）收集的房水中的TGF-β3进行定量。为开发TGF-β3 ELISA，我们筛选抗体以确定最佳组合，验证该检测方法在房水中的适用性，并优化制备活化TGF-β3的程序。结果，我们的ELISA具有高度的选择性和可重复性。使用我们的ELISA，我们发现PE患者眼中TGF-β3的浓度显著升高。我们还开发了新的TGF-β1和-β2 ELISA，并首次能够对单眼房水中的所有TGF-β亚型进行定量，以评估它们对眼部疾病发病机制的比例影响。
