Evaluation of the effectiveness of virus removal during the purification of immunoglobulin G by chromatography

Introduction. An innovative manufacturing process was developed to produce the highly purified preparation BioGam® "Human Immunoglobulin Normal, solution for infusion, 50 mg / ml, 100 mg / ml" using a combination of ethanol fractionation and chromatography on ion exchange and hydrophobic resin and an additional virus inactivation steps to achieve a reliable level of preparation safety. Aim. This study aimed to investigate the nucleic acids (NA) clearance efficacy of the hepatitis B (HBV), hepatitis C viruses (HCV) and parvovirus B19 (B19 V) during chromatography process for the isolation and purification of immunoglobulin G from precipitate A (Cohn fraction II + III). Materials and methods. Investigations were performed in model experiments with 3 series of precipitate A (II+III) samples knowingly contaminated with a virus containing material in which NA concentration of HBV virus was (1.77 ± 0.42) · 104 IU / ml, HCV virus was (1.42 ± 0.14) · 103 IU / ml, B19V virus (1.82 ± 0.14) · 105 IU / ml and which were exposed to chromatographic separation on the scale-down model of the manufacturing process. The virus reduction factor was determined by the change in viral load in product fractions before and after the chromatography process. Results. It has been established that the chromatography process demonstrated a reliable and reproducible level of virus nucleic acids reduction including to an undetectable by PCA level providing a reduction factor for HBV (3.61 ± 0.11) lg, for HCV (2.23 + 0.03) lg and for B19V (3.77 ± 0.02) lg. Conclusion. A chromatographic stage in the manufacturing process of preparation BioGam® makes a significant contribution to ensuring its viral safety wherein the removal efficiency of blood-transmissible agents has been confirmed experimentally.

immunoglobulin G, chromatography, scale-down model of manufacturing process, viral reduction, blood preparations