The Study of the Preparation of Purified Fibronectin from the Human Plasma.

Kim, Jae Chan; Kim, In Sik; Kim, Hack Chul; Shyn, Kyung Hwan; Koo, Bon Sool; Lee, Hee Sung

Original Article

Journal of the Korean Ophthalmological Society 1989;30(4):189-197.
Published online January 1, 2001.

The Study of the Preparation of Purified Fibronectin from the Human Plasma.

Jae Chan Kim, In Sik Kim, Hack Chul Kim, Kyung Hwan Shyn, Bon Sool Koo, Hee Sung Lee

¹Department of Ophthalmology, Medical School, Chung Ang University, Korea.
²Department of Biochemistry, Medical School, Chung Ang University, Korea.

Abstract

Fibronectin(FN), a glycoprotein present in plasma and extracellular matrix, has been reported to be effective on various corneal disorders such as persistent epithelial defect, corneal trophic ulcer, herpetic keratitis, etc. We performed the study of the preparation of purified FN from 35 persons' plasma(male: 19, female: 16)by the use of Sepharose 48 Affinity Chromatography and Gel filtration. The prepared FN was pure electrophoretically, and no other plasma proteins were contaminated, and confirmed as pure by multiple methods such as SDS-P AGE, Ouchterlony double gel diffusion and immunoelectro phoresis. Only 40 minutes was taken for preparation. The result showed that FN concentrations in plasma and in prepared solution were 288.9 +/- 72.8 microgram/ml and 294.3 +/- 41.4 microgram/ml respectively. FN concentrations in plasma and FN eyedrops showed no difference between sex, but increased their level with age(p<0.05). The biological activity was better preserved at 4 degrees C(in the refrigerator) than at room temperature till about 4 weeks after preparation. Applicability of this method as a useful preparation of purified FN is recommendable.

Key Words: Affinity chrmatography;Biologic activity;Fibronectin;Fibronectin concentration;Purification