Peptide affinity chromatography process for adsorption of fibrinogen

Fibrinogen is a 340 kDa protein molecule also known as Factor I. Its synthesis occurs in the liver of animals, In human beings around 2.6 g/l fibrinogen is found in the circulatory plasma playing a very important role in the blood coagulation. Due to the biological functions and characteristics of fibrinogen, the development of new techniques for separating and purifying fibrinogen from human plasma is a current challenge. Affinity chromatography based on peptide libraries has become known as a powerful technique for dealing with protein molecules, The lack of many important parameters involved in such systems has Limited the scale up. In this paper a way to obtain parameters such as the adsorption rate constants, by fitting the breakthrough curves, is presented. Dynamic experiments were carried out on HPLC (High Performance Liquid Chromatography) columns packed with resins, when the peptide FLLVPL was immobilized. This modified resin was also used for equilibrium experiments. This set of experiments allowed us to determine the maximum binding capacity, the dissociation constant and the lumped and intrinsic forward rate constants for resins with two different peptide densities (41.3 and 67.7 mu mol of peptide/g resin). The mathematical model describing the breakthrough curve uses the Langmuir type isotherm, and the axial dispersion was ignored. (C) 1999 Elsevier Science S.A. All rights reserved.