J Korean Ophthalmol Soc.  1989 Aug;30(4):189-197.

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

• Kim JC
• Kim IS
• Kim HC
• Shyn KH
• Koo BS
• Lee HS

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.

Keyword

Affinity chrmatography; Biologic activity; Fibronectin; Fibronectin concentration; Purification

MeSH Terms

Blood Proteins
Chromatography, Affinity
Chromatography, Gel
Diffusion
Extracellular Matrix
Female
Fibronectins*
Glycoproteins
Humans*
Keratitis, Herpetic
Ophthalmic Solutions
Plasma*
Sepharose
Ulcer
Blood Proteins
Fibronectins
Glycoproteins
Ophthalmic Solutions
Sepharose