![]() ![]() Namely, the mixed-mode resins have at least part of the Protein A properties in terms of their multiple binding mechanisms. Mixed-mode resins that combine hydrophobic properties with electrostatic properties for binding or elution have shown promising results. To address these issues, attempts have been made to replace expensive Protein A resin with conventional chromatographic resins. Other problems include the low binding capacity of Protein A resin and Protein A leaching. Thus, the cost of using Protein A depends on how many cycles the ligand can be reused. At the commercial scale, the cost of Protein A is a major issue, as low pH elution can cause damage not only to the antibodies to be purified but also the Protein A ligand, which can reduce the lifetime of the resin. With this method, antibody binding, which may potentially cause protein denaturation or aggregation, is not involved. We previously showed that agarose native gel electrophoresis can separate antibodies and host cell contaminants effectively, and hence here investigated the feasibility of antibody purification using native gel electrophoresis. Here, we review methods of antibody purification that do not use a Protein A column. In addition, such a harsh elution condition can cause the leaching of Protein A, which can impact in vivo effects of the eluted antibodies. This acid elution can cause denaturation and hence a loss of potency. In the research setting, the major problem with Protein A chromatography is not the high cost of Protein A resin but the difficulty in elution due to strong protein–protein interactions between antibodies and the Protein A ligand, which requires strong acid for reasonable recovery. The process of antibody production at both small and large scales has been comprehensively reviewed. Protein A chromatography constitutes a purification platform technology, although expensive, for research and the commercial-scale production of antibodies. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |