フィブロネクチン点眼剤調製の自動化への試み

Abstract

To develop an automatic device for the preparation of fibronectin eyedrops a basic study was performed to establish the optimum conditions for the preparation procedures. Fibronectin eyedrops were prepared according to the method of Nishida et al. In the first step with gelatinsepharose 4B (GS4B), the optimum conditions were as follows:(1) At least five and 15 ml of phosphate buffered saline (PBS) containing citrate were needed to wash the GS4B before and after plasma treatment, respectively;(2) The economical optimum ratio of GS4B vs. human plsama was 1: 10;(3) Preparations at either low or high temperatures should be avoided (optimum temperature 22°C); and (4) Higher yields of fibronectin were obtained with slower plasma flow rates. In the second step with sephadex-G25 (PD-10):(1) 20 ml of PBS were needed for pre-washing of PD-10; and (2) A high concentration and high yield fraction of fibronectin could be obtained when the eluate of 4 M urea from GS4B containing fibronectin was directly infused into PD-10. These results are considered important for the effective preparation of fibronectin eyedrops either automatically or manually.