Recombinant Protein A Immobilized on Cross-linked Cellulose Microspheres for Immunoglobulin G Adsorption from Human Plasma

Abstract

Cross-linked cellulose microspheres (CL-CMs) were successfully prepared using an inverse crosslinking suspension method from a cellulose solution with sodium hydroxide/urea aqueous solution as a solvent and epichlorohydrin as the crosslinker. The effects of epichlorohydrin content on the appearance and dispersity, average pore volume, moisture content, and wet real density of CL-CMs were studied. The microspheres presented a good spherical shape and porous surface structure. After activation with NaIO4, the recombinant protein A was immobilized onto the surface of CL-CMs to form an immunoadsorbent. Adsorbents containing various amounts of protein A were applied to adsorb immunoglobulin G (IgG) from human plasma. The maximum IgG adsorption capacities with static adsorption and dynamic adsorption were 23 and 13 mg, respectively, per gram of CL-CMs carrying 6.8 mg of recombinant protein A. Therefore, CL-CMs immobilized with recombinant protein A have great potential for application in the field of blood purification.

Affinity Resins for the Isolation of Immunoglobulins G Obtained Using Biocatalytic Technology

“…This has been achieved due to the site-specific reaction, which is an undoubtful advantage when compared to other chemical immobilization methods. Thus, the application of some nonspecific chemical methods frequently results in the formation of resins with dynamic capacity which do not exceed 13 mg/mL [16]. Alternatively, Wang and co-authors demonstrated that the introduction of the glutaraldehyde as an additional linker and a change in the immobilization conditions can alter to some extent the protein orientation on the matrix and increase DBC 10 values (up to 45 mg of IgG per 1 mL of the resin) in resulting resins [17].…”