Preparation of Placental (Fetal Tissue) Fibronectin and its Carbohydrates

Roger A. Laine
Susan J. Fisher
Betty C.R. Zhu

Abstract

This chapter describes the method used in laboratory for the preparation of human placental (fetal) tissue fibronectin. A major advantage of this technique is the relatively large amount of cell- and basement membrane-associated fibronectin which can be rapidly isolated: 50 to 120 mg of purified glycoprotein is purified from a single term placenta. Methods for isolating fibronectins are based on the affinity of all the glycoproteins for denatured collagen. Thus, the first step toward purification is usually chromatography on gelatin-Sepharose. Gel permeation chromatography of the bound fraction results in a relatively homogeneous protein preparation as demonstrated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The cost of purchasing sufficient heparin and DTT to extract additional fibronectin from the urea-insoluble residue from one placenta is about $200. If fibronectin is isolated on a routine basis, a simple, economic alternative is to process additional placentas. Because 50 mg of fibronectin can be extracted from only one placenta using urea alone the procedure can easily be “scaled-up” by the processing of several placentas simultaneously. © 1987, Elsevier Inc. All rights reserved.