Preparation and properties of an alpha‐1‐protease inhibitor concentrate with high specific activity

Mattes, E.; Matthiessen, H. Peter; Turecek, Peter L.; Schwarz, Hans‐Peter

doi:10.1046/j.1423-0410.2001.00049.x

Cited by 15 publications

(6 citation statements)

References 45 publications

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“…The separation membrane prevented CPV from migrating into the product stream and at least a 4 log reduction was demonstrated with PCR and infectivity assays ( Table 1). The high yield and 10-fold reduction of albumin in Gradiflow a-1-antitrypsin exceeds current multistep purification schemes that achieve 15-63% yields [22,23] with 60-90% purity, after as many as seven separate purification steps. Similar to commercial IgG processes, Cohn precipitation (Cohn IV-I) and PEG precipitation affect the yield and quality of the commercial a-1-antitrypsin product.…”

Section: Purification and Decontamination Of Ae-1-antitrypsinmentioning

confidence: 89%

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Blood protein purification and simultaneous removal of nonenveloped viruses using tangential-flow preparative electrophoresis

Evtushenko

Wang²,

Stokes

et al. 2005

Electrophoresis

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Gradiflow is new technology allowing purification of important blood proteins from viral contaminated plasma. Protein purification is based on unique scalable tangential-flow preparative electrophoresis, and is distinct from current technology because protein purification and virus removal are performed in the same step. This one-step removal and purification exploits both the size and charge of target proteins. The medically important blood proteins, immunoglobulin G (IgG) and alpha-1-antitrypsin, were chosen to demonstrate the ability of this process to purify proteins from contaminated plasma. Clearance factors achieved by infectivity assays and polymerase chain reaction (PCR) that meet regulatory requirements demonstrated removal of canine parvovirus (CPV). CPV is a model virus for pathogenic nonenveloped viruses, including parvovirus B19, not adequately removed or inactivated by most processes currently in practice. The recovery of proteins from plasma with high purity, recovery, and function, while simultaneously removing viruses, provides blood products with a level of purity compatible with clinical use more quickly and cheaply than available techniques.

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Section: Purification and Decontamination Of Ae-1-antitrypsinmentioning

confidence: 89%

“…Immunoglobulins are traditionally isolated using Cohn ethanol precipitation [20] followed by ion-exchange chromatography polishing steps. a-1-Antitrypsin [21][22][23] is an acid glycoprotein of 53 kDa with a pI of 4.8, and is widely used in the treatment of hereditary emphysema. Conventional purification…”

Section: Introductionmentioning

confidence: 99%

Blood protein purification and simultaneous removal of nonenveloped viruses using tangential-flow preparative electrophoresis

Evtushenko

Wang²,

Stokes

et al. 2005

Electrophoresis

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“…AAT is the most abundant serine protease inhibitor found in the blood , and it possesses the ability to prevent connective tissues from degrading by elastase. AAT has the capacity to inhibit 90% of trypsin in the human plasma , and it is normally used to cure lung emphysema induced by hereditary AAT deficiency. It was reported that more than 1 million people have this plasma protein deficiency .…”

Section: Introductionmentioning

confidence: 99%

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

Huangfu

Zhang

et al. 2017

Biotech and App Biochem

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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 log and 5.38 log were achieved for PRV and VSV, respectively. Porcine parvovirus and encephalomyocarditis virus were inactivated by 3.17 log and 5.88 log 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.

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“…a1-PI rapidly inactivates neutrophil elastase and therefore reduced tissue proteolysis. a1-PI deficiency can result in lung emphysema in adults, liver disease in children and is also associated with cystic fibrosis, arthritis and other malignant conditions (Huang et al, 2001;Mattes et al, 2001;Travis et al, 1985). Human plasmaderived a1-PI is a FDA----licensed product used for replacement therapy.…”

Section: Introductionmentioning

confidence: 99%

Production, purification, and characterization of human α1 proteinase inhibitor from Aspergillus niger

Chill

Trinh

Azadi

et al. 2008

Biotech & Bioengineering

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Human alpha one proteinase inhibitor (alpha1-PI) was cloned and expressed in Aspergillus niger, filamentious fungus that can grow in defined media and can perform glycosylation. Submerged culture conditions were established using starch as carbon source, 30% dissolved oxygen concentration, pH 7.0 and 28 degrees C. Eight milligrams per liter of active alpha1-PI were secreted to the growth media in about 40 h. Controlling the protein proteolysis was found to be an important factor in the production. The effects of various carbon sources, pH and temperature on the production and stability of the protein were tested and the product was purified and characterized. Two molecular weights variants of the recombinant alpha1-PI were produced by the fungus; the difference is attributed to the glycosylated part of the molecule. The two glycoproteins were treated with PNGAse F and the released glycans were analyzed by HPAEC, MALDI/TOF-MS, NSI-MS(n), and GC-MS. The MALDI and NSI- full MS spectra of permethylated N-glycans revealed that the N-glycans of both variants contain a series of high-mannose type glycans with 5-20 hexose units. Monosaccharide analysis showed that these were composed of N-acetylglucos-amine, mannose, and galactose. Linkage analysis revealed that the galactosyl component was in the furanoic conformation, which was attaching in a terminal non-reducing position. The Galactofuranose-containing high-mannnose type N-glycans are typical structures, which recently have been found as part of several glycoproteins produced by Aspergillus niger.

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Preparation and properties of an alpha‐1‐protease inhibitor concentrate with high specific activity

Abstract: The high specific activity of the A1PI preparation achieved with this process should allow a reduction of the A1PI total protein load necessary to achieve clinically relevant effects.

Cited by 15 publications

References 45 publications

Blood protein purification and simultaneous removal of nonenveloped viruses using tangential-flow preparative electrophoresis

Blood protein purification and simultaneous removal of nonenveloped viruses using tangential-flow preparative electrophoresis

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

Production, purification, and characterization of human α1 proteinase inhibitor from Aspergillus niger

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