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

Simona Viglio,Jan Stolk,Paolo Iadarola,Maura D’Amato

doi:10.3390/molecules25174014

Simona Viglio, Jan Stolk + Show 2 more

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https://doi.org/10.3390/molecules25174014

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Abstract

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.

Highlights

Alpha1-antitrypsin (AAT) is an acute-phase glycoprotein whose main function is to protect tissues from the enzymes of inflammatory cells, especially human neutrophil elastase (HNE) released by neutrophils during an inflammatory state [1,2,3,4,5,6]
Protein purification may range from a simple one-step procedure to complex approaches that involve more than one step
The results were poor in terms of both protein purity and quantity, the small amounts of AAT recovered being still heavily contaminated by interfering proteins

Summary

Introduction

Alpha1-antitrypsin (AAT) is an acute-phase glycoprotein whose main function is to protect tissues from the enzymes of inflammatory cells, especially human neutrophil elastase (HNE) released by neutrophils during an inflammatory state [1,2,3,4,5,6]. AAT is a 52 kDa single-chain protein synthetized as a 418 amino acid precursor that migrates on SDS-PAGE as a single band with an approximate Mr of 52 kDa (lanes 2 and 3 of Figure 1, panel A). Alpha1-antitrypsin (AAT, database entry ID P01009 according to UniProt) and the AAT–HNE (human neutrophil elastase) complex are represented as single bands with an approximate Mr of 52 kDa and 80 kDa, respectively (unpublished data from our laboratory). The reactive center loop (RCL) in the upper pole of the molecule shows the P1–P10 residues (Met358 and Ser359)

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