ALTERNATIVAS CROMATOGRÁFICAS PARA LA REMOCIÓN DE ENDOTOXINAS RESIDUALES EN EL PROCESO DE PURIFICACIÓN DE INMUNOGLOBULINAS HUMANAS

Abstract

Two ion exchange chromatographic alternatives in negative mode were evaluated, on a laboratory scale, for the removal of endotoxins in immunoglobulin solutions. Was analyzed the influence of three technological parameters (protein concentration, ionic strength and pH) on the behavior of the two main technical efficiency criteria of the procedure: 1. Endotoxin removal capacity expressed as total EU (endotoxin units), 2. Protein recovery expressed as mg/total protein. The main goal of the work was to select variants that meet the regulatory requirement for intravenous immunoglobulins, which has to be < 1 EU/mL. The incidence of protein concentration and ionic strength against a known endotoxin load on the behavior of both technical efficiency criteria was evaluated for the chromatographic resin Fractogel-TMAE. The variant with a protein concentration close to 8%, without ionic strength reinforcement, was the best option. As a second alternative for a lower endotoxin load, was evaluated for a chromatographic membrane of the Sartobind-Q type, the incidence of pH on the behavior of both technical efficiency criteria, maintaining the protein concentration around ~8%. In this case the variant at pH=6.5 was the best integral option. For immunoglobulin solutions with protein concentration ≤ 8.0 %, both alternatives are effective up to 4 column volume, in terms of endotoxin removal capacity and recovery greater than 85 %, since both meet the regulatory requirement for intravenous immunoglobulins.