ESTABLISH PROCEDURE DETERMINATION OF MOLECULAR SIZE DISTRIBUTION OF HUMAN ALBUMIN AND HUMAN IMMUNOGLOBULIN PRODUCTS BY SIZE EXCLUSION HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Abstract

Quality control of human albumin and human immunoglobulin products is critical to ensuring product safety and efficacy. According to the European Pharmacopoeia and the US Pharmacopoeia, the test evaluation of the molecular size distribution of proteins in albumin and immunoglobulin products is a mandatory test and one of the most important in the quality assessment of proteins quantity of biological products. The European Pharmacopoeia stipulates that the content of undesirable impurities in albumin products such as polymers and aggregates should not exceed 5%. This study was conducted to develop a general procedure to determine the molecular size distribution of protein in human albumin and human immunoglobulin products in accordance with the actual conditions of the Faculty. The study used descriptive experimental method in the laboratory. The procedure for determining the molecular size distribution of protein in Human Albumin Baxter 200 g/l and IV Globulin products was performed on a Thermo Scientific HPLC chromatography system, using a TSK G3000SW Column (7.5 mm x 60 cm) and DAD detector with optimized parameters: sample concentration 1%, standard concentration 1%, mobile phase concentration: 40 mM phosphate buffer, 200 mM NaCl, 0.005% sodium azide, flow rate 0, 5 ml/min. The procedure has been validated with system suitability, precision, robustness, specificity, and the uncertainty of  Monomer + Dimer and Polymer + Aggregate in Human Albumin Baxter 200 g/l are respective 0,1285% and 0,1285%, in IV Globulin are respective 0.0463% and 0.0255% (with 95% confidence). The study has successfully provided a general procedure for determining the molecular size distribution of protein in human albumin and human immunoglobulin products, the procedure has been validated and is suitable for practical conditions at Faculty.