Chromatographic purification of human α1 proteinase inhibitor from dissolved Cohn fraction IV-1 paste

Chen, Sharon X.; Hammond, David J.; Klos, Anthony M.; Wood, Woody D; Wydick, James E.; Lebing, Wytold R.

doi:10.1016/s0021-9673(97)01119-9

Cited by 14 publications

(4 citation statements)

References 20 publications

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“…It has been asserted that the isolation of A1PI by cation‐exchange chromatography leads to a dramatic improvement in purity [33,45,46]. At a pH lower than 6·0, the active A1PI does not bind to the cationic column, whereas contaminating proteins are bound.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2001

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Section: Discussionmentioning

confidence: 99%

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

et al. 2001

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“…The purification of AAT from Cohn Fraction IV was described also by Chen et al [ 38 ], who followed an articulated procedure consisting of a combination of complementary chromatographic steps. They coupled anion to cation exchange chromatography used as the first and second step, respectively.…”

Section: Methods Of Aat Purification: From the Beginning To The Prmentioning

confidence: 99%

Methods of Purification and Application Procedures of Alpha1 Antitrypsin: A Long-Lasting History

2020

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The aim of the present report is to review the literature addressing the methods developed for the purification of alpha1-antitrypsin (AAT) from the 1950s to the present. AAT is a glycoprotein whose main function is to protect tissues from human neutrophil elastase (HNE) and other proteases released by neutrophils during an inflammatory state. The lack of this inhibitor in human serum is responsible for the onset of alpha1-antitrypsin deficiency (AATD), which is a severe genetic disorder that affects lungs in adults and for which there is currently no cure. Being used, under special circumstances, as a medical treatment of AATD in the so-called “replacement” therapy (consisting in the intravenous infusion of the missing protein), AAT is a molecule with a lot of therapeutic importance. For this reason, interest in AAT purification from human plasma or its production in a recombinant version has grown considerably in recent years. This article retraces all technological advances that allowed the manufacturers to move from a few micrograms of partially purified AAT to several grams of highly purified protein. Moreover, the chronic augmentation and maintenance therapy in individuals with emphysema due to congenital AAT deficiency (current applications in the clinical setting) is also presented.

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“…a1-PI exhibits a molecular mass of approximately 52 kDa, has 394 amino acids, a single cystein residue and three carbohydrate attachment sites at asparagine residues 46, 83, and 247 (Carrell et al, 1982;Chen et al, 1998;Kolarich et al, 2006a,b). Significant research efforts were directed to produce recombinant human a1-PI in E. coli (Courtney et al, 1984), yeast (Kwon et al, 1995), transgenic mice (Zbikowska et al, 2002) and sheep (Wright et al, 1991) as well as plant cells (Huang et al, 2001).…”

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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Chromatographic purification of human α1 proteinase inhibitor from dissolved Cohn fraction IV-1 paste

Cited by 14 publications

References 20 publications

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

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

Methods of Purification and Application Procedures of Alpha1 Antitrypsin: A Long-Lasting History

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

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