Ion-exchange chromatography separation of the detergent and the solvent from immunoglobulins after solvent-detergent treatment.

Abstract For inactivation of lipid-enveloped viruses during the immunoglobulin production, the solvent–detergent (S/D) method was applied. Tri- n -butyl phosphate (solvent) and Triton X-100 (detergent) were removed from S/D treated immunoglobulins by ion-exchange chromatography on Q-Sepharose Fast Flow (FF). During the chromatographic procedure immunoglobulins remained bound on a Q-Sepharose FF, whereas solvent and detergent were eluted by washing with starting buffer. Elution of immunoglobulins was achieved by increasing the ionic strength of the starting buffer. The final immunoglobulin preparation contained less than 10 μg/ml of Triton X-100 and less than 2 μg/ml tri- n -butyl phosphate. It was confirmed that the S/D procedure did not cause a significant change in polymers and specific antibodies content. Immunoglobulin classes were also not affected by the same procedure.