Collaborative Study for the Establishment of National Reference Standard for Molecular Size Distribution Test of Human Immunoglobulin Products

Abstract

Human immunoglobulin is a plasma-derived medicinal product that is the second-largest product in the numberof the national lot release in the National Institute of Food and Drug Safety Evaluation (NIFDS), following human serumalbumin. Reference standards are used to confirm the validity of each national lot release test. This study aims to establisha national reference standard (NRS) for the molecular size distribution test applied to immunoglobulin based biologics. 7200 vials, containing 2 mL/vial, were prepared according to the approved manufacturing process for humanimmunoglobulin intended for intramuscular administration. Four laboratories, including the NIFDS, and manufacturers ofhuman immunoglobulin products, participated in this collaborative study, in which a molecular size distribution test wasperformed on candidate vials using size-exclusion high-performance liquid chromatography. Retention of dimer relative tomonomer was 0.851. Relative to a human immunoglobulin (molecular size) biological reference preparation, relativeretention times of monomer and dimer from the candidate vial were 1.001 and 1.000, respectively. This result satisfies therequirements of monographs in the European Pharmacopoeia. Based on a real-time stability evaluation over 24 monthsfollowing manufacture, and a short-term stability evaluation over 14 days after opening, the dimer to monomer relativeretention ratios were found to be distributed within control limits, thereby validating stability. The results of this studysupport registration of this candidate with the NIFDS as the NRS for molecular size distribution test of humanimmunoglobulin products.