Large‐scale purification of high purity α1‐antitrypsin from Cohn Fraction IV with virus inactivation by solvent/detergent and dry‐heat treatment

α1‐Antitrypsin (AAT) is widely used to treat patients with congenital AAT deficiency. Cohn Fraction IV (Cohn F IV) is normally discarded during the manufacturing process of albumin but contains approximately 33% of plasma AAT. We established a new process for large‐scale purification of AAT from it....

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Bibliographic Details
Published in:Biotechnology and applied biochemistry 2018-05, Vol.65 (3), p.446-454
Main Authors: Huangfu, Chaoji, Zhang, Jinchao, Ma, Yuyuan, Jia, Junting, Li, Jingxuan, Lv, Maomin, Ma, Xiaowei, Zhao, Xiong, Zhang, Jingang
Format: Article
Language:English
Subjects:
a1-antitrypsin
Blood plasma
blood product
Chromatography
Cohn Fraction IV
Deactivation
Freeze drying
Heat
Heat treatment
Inactivation
Ionization
Ions
Liquid chromatography
Manufacturing industry
Mass spectrometry
Mass spectroscopy
Parvoviruses
plasma protein
Purification
Purity
Qualitative analysis
Solvents
Stomatitis
viral safety
Viruses
α1‐antitrypsin
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Summary:α1‐Antitrypsin (AAT) is widely used to treat patients with congenital AAT deficiency. Cohn Fraction IV (Cohn F IV) is normally discarded during the manufacturing process of albumin but contains approximately 33% of plasma AAT. We established a new process for large‐scale purification of AAT from it. liquid chromatography‐electrospray ionization‐tandem mass spectrometry and high‐performance liquid chromatography were applied for qualitative identification and composition analysis, respectively. Stabilizers were optimized for AAT activity protection during lyophilization and dry‐heat. Virus inactivation by dry‐heat and solvent/detergent (S/D) was validated on a range of viruses. AAT with purity of 95.54%, specific activity of 3,938.5 IU/mg, and yield of 26.79%, was achieved. More than 95% activity was reserved after S/D. More than 96% activity was obtained after lyophilization or dry‐heat. After S/D, pseudorabies virus (PRV) and vesicular stomatitis virus (VSV) were inactivated below detectable level within 1 H. Virus titer reductions of more than 5.50 log10 and 5.38 log10 were achieved for PRV and VSV, respectively. Porcine parvovirus and encephalomyocarditis virus were inactivated by 3.17 log10 and 5.88 log10 reduction after dry‐heat. The advantages of this process, including suitability for large‐scale production, high purity, better utilization of human plasma, viral safety, commercial and inexpensive chromatography medium, may facilitate its further application.
ISSN:0885-4513
1470-8744
DOI:10.1002/bab.1623