Variability of tear protein levels in normal young adults: Between-day variation

Background: An adequate knowledge of physiological variation is important for valid comparative studies of tear proteins. The aim of this study was to investigate the between-day variation of the human tear protein levels, including the total protein concentration (TPC) and the levels of major protein fractions. Two sampling methods, the yawn and the eye-flush methods, were used and compared. Methods: TPC was determined by the Bradford method. The major protein fractions were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and their levels were determined by scanning densitometry after SDS-PAGE. The tear protein levels were monitored for 3 days. Results: The between-day differences in the levels of TPC and the individual protein fractions were not statistically significant in either sampling method, but the variations of some proteins were large and would be clinically significant. Different variations were observed in different proteins. The variations in serum albumin were large, up to 61% and 113% in the yawn and eye-flush methods respectively. The variations in lactoferrin, tear-specific prealbumin and lysozyme were relatively small in the yawn method. The variations in protein levels obtained by the eye-flush method were generally higher than by the yawn method. Conclusion: Although the between-day differences in tear protein levels were not statistically significant, the variations in some proteins would be large in magnitude. The variability of tear protein levels obtained by the eye-flush method was larger than that by the yawn method. Therefore, caution should be taken if the eye-flush method is used for sampling tears for quantitative analysis of tear proteins, although it is easier to collect tear samples using this method. The results will be useful to exclude normal variation in tear protein levels when comparing pre- and post-therapeutic tear protein levels in eyes treated for tear-related abnormalities.