Disease-related quantitation of TGF-beta3 in human aqueous humor

TGF-beta 3 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-beta 3 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-beta 3 ELISA, we screened antibodies to identify the best combination, validated the assay for aqueous humor, and optimized the procedure for preparing activated TGF-beta 3. As a result, our ELISA was highly selective and reproducible. Using our ELISA, we discovered a significantly elevated concentration of TGF-beta 3 in PE eyes. We also developed new TGF-beta 1 and - beta 2 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.