Isolation and preliminary characterization of tear prealbumin from human ocular mucus

Abstract

Tear prealbumin was purified from crude tear prealbumin previously isolated from the saline soluble human ocular mucus. Purification was achieved by further column chromatographies on DEAE Sephadex A-25 and Sephadex G-75. Preliminary characterization included amino acid analysis, gel electrophoresis, and isoelectric focusing. Unlike serum prealbumin, the purified tear prealbumin showed a predominance of acidic residues and a trace amount of tryptophan. It exhibited polymorphic nature, with pi values of 4.8 and 4.9. The possibility of a tear prealbumin/retinol complex was also examined. The protein was found to incorporate with 3H retinol. The % retinol-incorporated tear prealbumin did not exhibit the characteristic UV spectrum of retinol; however, it did display emission and excitation fluorescence spectra at high concentrations similar to serum retinol-binding protein.