Protein A immunoaffinity hollow fiber membranes for immunoglobulin G purification: Experimental characterization

• Published in: Biotechnology and bioengineering 1995-11, Vol.48 (4), p.415-427
• Main Authors: Charcosset, Catherine, Su, Zhiguo, Karoor, Sujatha, Daun, Gregor, Colton, Clark K.
• Format: Article
• Language: English
• Subjects: affinity membranes; Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; IgG; immunoadsorption; Methods. Procedures. Technologies; Others; protein A; Various methods and equipments
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.260480414

Immunoaffinity adsorption is increasingly used for protein purification and medical applications. Synthetic membranes have advantages as support matrices in comparison to conventional bead supports because they are not compressible and they eliminate internal diffusion limitations. The goal of this study was to explore in detail the performance of microporous hollow fibers composed of modified polysulfone to which protein A was immobilized for adsorption of human IgG. The internal matrix was characterized by scanning electron microscopy. The binding equilibrium constant was measured using both static and dynamic methods. Break‐through curves up to ligand saturation were measured and used to study the effects of IgG concentration, presence of contaminant albumin, flow direction, flow mode, and especially filtrate flow rate and maximum IgG binding capacity. The highest binding capacities studied were comparable with that attainable with bead matrices. All of the breakthrough curves could be represented on a single figure when plotted versus the dimensionless relative throughput (the mass of IgG loaded on the membrane divided by the mass that would be bound when the entire fiber is in equilibrium with the feed concentration), and the effect of operating variables on the position and shape of the individual breakthrough curves could be understood in terms of a dimensional performance parameter (the product of membrane volume and maximum binding capacity divided by the filtrate flow rate). The best breakthrough curves were obtained with the highest values of the performance parameter. Based on the results, membranes as solid supports for immunoadsorption can be a useful alternative to the use of traditional columns for protein separations. © 1995 John Wiley & Sons, Inc.