Abstract：OBJECTIVE To establish a method for detecting the impurity components in human serum albumin by electrospray ionization mass spectrometry (ESI-Q-TOF MS) and investigate the impurity components in 28 batches of post-marketing albumin products from 27 companies. METHODS According to Chinese Pharmacopoeia′s General Principles 4121, gel filtration column (Hiprep16/60 SepharylS-200HR) was used to separate the protein components. The dimer and multimer components were collected, then desalted, concentrated and digested by trypsin. The peptide was eluted by BEH C18(2.1 mm×100 mm,130) column with mobile phase consisting of water (0.1% formic acid)-acetonitrile in gradient elution mode. Chromatography-mass spectrometry was used to determine the secondary sequences of the peptides and the searching software (PLGS3.0) was used to identify the impurity protein components in albumin. RESULTS A total of 23 impurity proteins were detected, among which apolipoprotein A-II, haptoglobin, and hempoexin, α-1B-glycoprotein and α-2-HS-glycoprotein were the impurity proteins common to all enterprise products; α-albumin (VE binding protein), N-acetyl-L-alanine amidase, haptoglobin-related proteins, hemoglobin (β subunit, α subunit), transthyretin and other proteins were found in the products of most companies. The number of heterologous proteins in each enterprise was between 8-17, and the numbers of impurity proteins in 27 companies were quite different. CONCLUSION This method can be used for the investigation of unknown components in human serum albumin, which provides a guidance for the optimization of human albumin production process.
柯兵兵, 谢育媛, 王德蓉, 曾浩, 杨娟, 郭江红. 人血白蛋白中杂蛋白的生物质谱分析[J]. 中国药学杂志, 2020, 55(2): 148-152.
KE Bing-bing, WANG De-rong, ZENG Hao, YANG Juan, GUO Jiang-hong. Analysis of Impurity Proteins by Biomass Spectrometry in Human Serum Albumin. Chinese Pharmaceutical Journal, 2020, 55(2): 148-152.
WANG J X. Blood Products(血液制品学)[M]. Beijing:People′s Medical Publishing House,1998,74:57.
[2]
JIANG J K,REN J M. Great attention should be paid to the adverse reactions caused by human albumin preparations . Chin J Hosp Pharm(中国医院药学杂志),1994,(10):476-477.
[3]
Ch.P(2015). Vol Ⅲ(中国药典2015年版.三部)[S]. 2015.
[4]
LIU L, LIU W F. The history and development of human albumin separation process . Chin J Blood Transfus(中国输血杂志), 2008, 21(4):323-326.
[5]
WANG D M. Progress on Isolation and Purification of Human Blood Albumin. Biotech World(生物技术世界杂志),2014(11):134.
[6]
WANG M Y, WANG P, ZANG H C. Advances of human albumin production and its application . J Pharm Res(药学研究), 2014,14(10):600-602.
[7]
LISBETH B J, JØRGEN D, BORGE T, et al. Identification and removal of polymer- and aggregate forming proteins in human plasma albumin preparations. Vox Sanguinis, 1994, 67(2):125-131.
[8]
LIN J J, NEYER J D, CARPENTER J F, et al. Aggregation of human serum albumin during a thermal viral inactivation step. Int J Biol Macromol, 2009,45(2):91-96.
[9]
ZHANG D,CHEN B. The formation trend and preliminary analysis of polymer in the process of human serum albumin. Chin J Blood Transfus(中国输血杂志),2016,29(9):909-912.
[10]
GUO J H, KE B B, JIANG H. Analysis of human albumin impurity protein by ultra performance liquid chromatography electrospray quadrupole time of flight mass spectrometry. Her Med(医药导报), 2019,4(15):223-226.
[11]
WASSELL J. Haptoglobin: function and polymorphism. Clin Laboratory, 2000, 46(11-12):547-552.
[12]
KAPRALOV A, VLASOVA II, FENG W, et al. Peraxidase activity of hemoglobin-haptoglobin complexes: coavalent aggregation and oxidative stress in plasma and macrophages. Biol Chem, 2009, 284(44):30395-30407.
[13]
GEORGE A. Trapp, plasma aluminum is bound to transferrin. Life Sci, 1983, 3(4):311-316.
[14]
FATEMI S J, KADIR F H, MOORE G R, et al. Aluminium transport in blood serum. Binding of aluminium by human transferrin in the presence of human albumin and citrate. Biochem J, 1991, 280(2):527-532.
[15]
EI Hage CHANING J M, HMADI M, Ha-DUONG N T. Uptake and release of metal ions by transferrin and interaction with receptor 1. Sci Direct, 2012, 1820(3):334-347.
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