Alpha1-antitrypsin Deficiency


Modification of the Manufacturing Process for the Plasma-derived Human Alpha1-proteinase Inhibitor Leading to Prolastin®-C


Wytold Lebing, MS,1 Michelle St Peter, BS,2 Charles Ndarathi, DVM, PhD,2 James Rebbeor, PhD2 JoAnn Hotta, PhD,3 and Deborah Barnette, MS2


1. Senior Director, Business Development, Talecris Biotherapeutics Inc., Research Triangle Park, North Carolina; 2. Process Development and Technology, Talecris Biotherapeutics Inc., Clayton, North Carolina; 3. Pathogen Safety, Talecris Biotherapeutics Inc., Research Triangle Park, North Carolina


William R Alonso, PhD,2


Abstract


Prolastin® (alpha1-proteinase inhibitor [human], alpha1-PI) has been used since 1988 in patients with alpha1-antitrypsin deficiency. It is derived from human plasma using a series of separation, purification and pathogen reduction techniques. This article discusses the development of a


high-purity alpha1-PI, Prolastin®-C, from human plasma for chronic augmentation and maintenance therapy in adults with emphysema due to deficiency of alpha1-proteinase inhibitor, through a modification of the Prolastin manufacturing process. Additional chromatographic purification and two new dedicated virus reduction steps (solvent/detergent treatment and 15nm nanofiltration) were incorporated into the manufacturing process. Bench-scale models were used to validate these manufacturing steps. Protein characterization confirmed the high


purity of Prolastin-C. Global virus reduction factors for the Prolastin-C process were ≥10.2 to ≥25.5 log10 for enveloped viruses and ≥12.7 to ≥13.7 log10 for non-enveloped viruses, with a minimum 6.0 log10 reduction of transmissible spongiform encephalopathy infectivity. Modification of the manufacturing process has resulted in a product with high purity, a higher concentration of alpha1-PI, and an excellent margin of pathogen safety.


Keywords Alpha1-antitrypsin, alpha1-proteinase inhibitor, Prolastin, Prolastin-C, purity, prion, protein, transmissible spongiform encephalopathy, virus


Disclosure: All authors are full-time employees of Talecris Biotherapeutics Inc. Acknowledgments: The authors would like to thank Susan Trukawinski, Woody Wood, David Ownby, Scott Cook, Chris Dadd, Chad Ennis, and Mark Chavez. Received: October 5, 2010 Accepted: December 9, 2010 Citation: US Respiratory Disease, 2010;6:22–6 Correspondence: Deborah Barnette, MS, Development & Technology, Talecris Biotherapeutics Inc., 8368 US 70 West, Clayton, NC 27520, US. E: Deborah.Barnette@talecris.com


Support: This study was sponsored by Talecris Biotherapeutics Inc. (Research Triangle Park, NC 27709, US). Editorial assistance was provided under the direction of the authors by Anne-Marie Manwaring, BSc, and Martin Kenig, DPhil, of PAREXEL and was supported by Talecris Biotherapeutics Inc.


Alpha1-proteinase inhibitor (alpha1-PI), a major serine protease inhibitor (serpin) synthesized principally in the liver, enters the lungs via the


bloodstream, where it functions as the primary inhibitor of the proteolytic enzyme neutrophil elastase. If not suppressed by


alpha1-PI, neutrophil elastase may destroy healthy pulmonary tissue and cause chronic irreversible lung damage. Individuals with the genetic


disorder alpha1-antitrypsin (AAT) deficiency have low serum levels of alpha1-PI and are therefore at an increased risk of developing emphysema, sometimes as early as in the fourth or fifth decade of life.


Prolastin® (alpha1-PI, human) has been used since 1988 in patients with AAT deficiency. Observational studies have shown that Prolastin slows the


decline in lung function1–4 and has an excellent safety profile.5–8 The manufacturing process of Prolastin, a plasma-derived protein, involves


purification of alpha1-PI from Cohn fraction IV-1 paste by precipitation, filtration and ion exchange chromatography. Several of the purification steps also remove viruses and transmissible spongiform encephalopathy (TSE) agents or prions.


22


This report summarizes the recently-approved modifications and improvements to the Prolastin manufacturing process that have led to


the development of an alpha1-PI product. This product, designated Prolastin®-C, has high purity, a higher concentration of alpha1-PI than Prolastin and an excellent margin of pathogen safety.


Materials and Methods


Comparison of the Manufacturing Processes The production of both Prolastin-C and Prolastin is identical at the beginning of the manufacturing process. It involves thawing and pooling of the plasma, precipitation of the fraction IV-1 paste with polyethylene glycol (PEG) treatment, then centrifugation (whereupon the paste is discarded) and depth filtration of the supernatant (see Figure 1). In the Prolastin-C process, there is an additional cation exchange chromatography purification step and improved ultrafiltration and diafiltration for further purification. Two new dedicated virus reduction steps (solvent/detergent aka S/D treatment and 15nm nanofiltration) replace the pasteurization step of the Prolastin process (see Figure 1).


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