Analysis of human tear fluid components, inhibiting protein adhesion to plastic surfaces

Experimental Eye Research
A KuizengaA Kijlstra

Abstract

In a previous paper we reported the presence of components in human tear fluid that block the interaction of proteins with plastic surfaces, interfering with tear protein ELISA and proposed the term coating inhibiting activity. The purpose of the study presented here was to further analyse these components. Coating inhibitory activity in human reflex tears was analysed by lectin affinity chromatography, using the agarose bound lectin Artocarpus integrifolia agglutinin (Jacalin), gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotting and Jacalin staining. For coating inhibitory activity assay in experimental tear samples, the binding of the protein Avidin-conjugated horseradish peroxidase to the polystyrene surface of ELISA micro-titer plate wells, preincubated with the experimental tear samples was measured. In addition, tears were incubated with scrapings of the ELISA plates used in the assay and with six different types of contact lenses (two rigid gas permeable and four hydrogel soft contact lenses) for analysis of adsorbed components. Lectin affinity chromatography of tears yielded a Jacalin-binding and a non-Jacalin-binding preparation, both exhibiting coating inhibitory activity but...Continue Reading

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