Rapid preparation of purified autologous fibronectin eyedrops from patient's plasma.

Fibronectin (FN), a glycoprotein present in plasma and the extracellular matrix, has been reported to be effective on corneal trophic ulcer. This study reported a rapid method for preparing purified FN eyedrops from a patient's own plasma. Plasma FN concentration in healthy Japanese was found to be 235 +/- 62 micrograms per ml(age 21-31 years old). Men had a higher plasma FN level than women (264 +/- 62 vs. 206 +/- 44 micrograms/ml). The FN eyedrops were prepared by gelatin-coupled Sepharose 4B affinity chromatography and gel filtration by Sephadex G-25 (PD-10). Biochemical and immunochemical analyses revealed that the prepared FN eyedrops contained electrophoretically pure FN and no other plasma proteins were detected. Average FN concentration in the eyedrops was 425 micrograms per ml and recovery rate was 54%. The FN eyedrops were found to be stable at room temperature up to 10 days in biological activity as far as we examined. The total time required to prepare the FN eyedrops was within 2 hours. Thus it is possible to apply them to outpatients as well as inpatients.