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Li C.-S.,Wuhan Biological Products Co. | Zhou Z.-J.,Wuhan Biological Products Co. | Hu Y.,Wuhan Biological Products Co. | Li T.-J.,Wuhan Biological Products Co. | And 3 more authors.
Chinese Journal of Biologicals

Objective: To control the quality of pilot purification procedure of IgG by chromatography through determination of plasma specific protein. Methods: The compositions of material plasma used for production and pilot purification were analyzed by immune scatter turbidimetry using Dade Behring BN ProSpec analyzer (BNP) and ancillary reagents, based on which a data bank of distributions of albumin (ALB), IgG, IgA, IgM, complement 3 (C3), C4, trnasferin (TRF), α1-antitrypsin (AAT), α2-macroglobin(A2M), haptoglobin (HPT), α1-acid glycoprotein (AAG), ceruloplasmin (CER), fibrinogen(FIB), antithrombin III (AT III), IgG1 IgG2, IgG3 and IgG4 was established. The removals of ALB, IgG, IgA, IgM, FIB and AAT as well as recovery rate of IgG at various steps of pilot purification procedure of three batches of plasma were monitored to verify the stability of the procedure. The determination results of IgG contents in three batches of bulks and final products prepared by the pilot purification procedure were compared with those by Kjeldahal method to verify the accuracy and precision in intermediate assay by various staff. The foreign matter contents and subclasses of bulks of IgG prepared by pilot purification procedure and cold ethanol fractionation were analyzed and compared. Results: Only the TRF, A2M and AT III contents in pooled material plasmid used for pilot purification procedure showed significant difference with those in the plasma for production (P < 0.05). Most of ALB and AAT were removed from the plasma used for pilot purification by A2P affinity chromatography, while the residual ALB, FIB and IgM by caprylate precipitation, and IgA and ALB by DEAE ion exchange chromatography. The total recovery rate of IgG was more than 70%, indicating high stability of the procedure. The CVs of determination results of IgG contents in bulk and final product prepared by pilot purification procedure were less than 10%, while the recovery rates were 90%-110%, indicating high accuracy and precision. The foreign matter contents of bulk prepared by pilot purification procedure were lower than those by cold ethanol fractionation, while the distribution of IgG subclass was similar to that of material plasma used. Conclusion: Plasma specific protein determination may be used for monitoring and optimization of purification procedure of plasma proteins. Source

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