Development of a method for obtaining immunoglobulins A and M to create a complex immunoglobulin drug
Abstract
The possibility for obtaining immunoglobulins A and M for create a complex immunoglobulin drug, which is used for the prevention and treatment of acute intestinal diseases have been investigated. The effectiveness of this drug has been demonstrated in the course of numerous clinical trials and proven over time, but a drug that meets all modern requirements for purity and viral safety is not produced in our country. Previously, we developed a technology for obtaining virus-free immunoglobulin G, including the purification of inactivated blood plasma on three sorbents: hydrophobic, DEAE and sulfocationite. Studies have shown that immunoglobulins A and M are concentrated on the DEAE sorbent. After the introduction of the technology for obtaining immunoglobulin G, the issue of extracting the fraction of immunoglobulins A and M became especially relevant, since it can allow their isolation in one technology with the production of immunoglobulin G, which reduces economic costs and allows for a comprehensive solution to the problems of processing blood plasma. The work studies the conditions for elution of the fraction of immunoglobulins A and M for subsequent purification on various sorbents. It was shown that elution from a DEAE column with 0.075 M sodium chloride solution at pH 5.65 allows removing more than 60% of polymers in the raw material. Subsequent purification was carried out on six different sorbents that differed in the carrier (agarose and polymethacrylate) and functional groups (sulfonic cation exchanger, weak and strong anion exchanger) in four buffer systems with pH 5.0; 5.65; 6.8; and 7.4. It was shown that on the anion-exchange sorbent based on methacrylate Relisorb DA 400 at pH 7.4, a high yield of immunoglobulins is achieved (70% for IgA and 80% for IgM) with a low polymer content, which allows obtaining a complex immunoglobulin preparation that meets modern requirements.
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