Large-scale purification of high purity α1-antitrypsin from Cohn Fraction IV with virus inactivation by solvent/detergent and dry-heat treatment.
Biotechnol Appl Biochem
; 65(3): 446-454, 2018 May.
Article
in En
| MEDLINE
| ID: mdl-29072333
ABSTRACT
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Solvents
/
Blood Proteins
/
Alpha 1-Antitrypsin
/
Virus Inactivation
/
Detergents
/
Hot Temperature
Type of study:
Qualitative_research
Limits:
Animals
Language:
En
Journal:
Biotechnol Appl Biochem
Journal subject:
BIOQUIMICA
/
BIOTECNOLOGIA
Year:
2018
Document type:
Article