Determination and Quality Evaluation of Glycoprotein in Recombinant Human Albumin from Different Expression Systems / 中国药学杂志

ABSTRACT

OBJECTIVE: To determine glycoprotein content in recombinant human albumin from different expression systems with different methods. METHODS: Recombinant human albumin samples from Saccharomyces cerevisiae expression system, Pichia pastoris expression system, Oryza sativa expression system as well as plasma derived human albumin were investigated by phenol sulfuric acid method, HPLC peak area method and ConA combining elution HPLC method. RESULTS: For 10 batches of samples expressed in pichia pastoris expression system, the total contents of mannose were 2.7 mg•g(Pro)-1 (A manufacturer, n = 4) and 1.7 mg•g (Pro)-1 (B manufacturer, n = 6), respectively. The HPLC peak area percentages of ConA binding protein in recombinant human albumin from Pichia pastoris expression system were the highest, which showed 2.65% (A manufacturer) and 0.71% (B manufacturer) respectively, the peak area percentage of ConA binding protein in Oryza sativa expression system was 0.05% (E manufacturer, n = 3), and the ConA binding protein peak area of plasma derived human albumin was 0.01% (W manufacturer). The results of ConA binding and elution analysis with HPLC method for Quantitative determination of ConA binding protein showed that the ConA binding protein contents in the samples from pichia pastoris expression system were much higher 27.58 mg•g(Pro)-1 (A manufacturer), 21.48 mg•g(Pro)-1 (B manufacturer), 32.02 mg•g(Pro)-1 (C manufacturer); the ConA binding protein content in the sample from Saccharomyces cerevisiae expression system was lower, 2.29 mg•g(Pro)-1 (D manufacturer); the ConA binding protein content in the samples from oryza sativa expression system was the lowest, 1.27 mg•g(Pro)-1 (E manufacturer); the plasma derived human albumin ConA binding protein content was 31.16 mg•g(Pro)-1 (S manufacturer). CONCLUSION: In terms of the results of the samples and methods involved in this study, there were glycosylated or glycol forms of protein in all recombinant human albumin samples from different expression systems; the glycosylated protein content in samples of Pichia pastoris expression system is higher than Saccharomyces cerevisiae expression system, while the glycoformed protein in samples of Oryza sativa expression system is the lowest. Plasma derived human albumin also contains glycoprotein or glycosylated protein.