Human plasma‐derived immunoglobulin G fractionated by an aqueous two‐phase system, caprylic acid precipitation, and membrane chromatography has a high purity level and is free of detectable <i>in vitro</i> thrombogenic activity

Vargas, Mariángela; Segura, Álvaro; Wu, Yue; Herrera, María; Chou, Ming Li; Villalta, Mauren; León, Guillermo; Burnouf, Thierry

doi:10.1111/vox.12209

Cited by 13 publications

(18 citation statements)

References 32 publications

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“…Prothrombin time (PT) and activated partial thromboplastin time (aPTT) analysis was performed on a Sysmex CA‐1500 system. Thrombin generation assay (TGA) was performed using a TGA kit (Technothrombin, Technoclone) as described previously . Commercial controls were spiked at 1% (vol/vol) and test samples at 10% (vol/vol) into platelet‐poor plasma.…”

Section: Methodsmentioning

confidence: 99%

“…Fluorescence was read at 360/460 nm for 60 minutes at 1‐minute intervals at 37°C in a microplate fluorescence reader (Varioskan Flash, Thermo Scientific). Proteolytic activity assays to detect plasmin, FXa, and thrombin‐like activity were from Chromogenix . Control plasma spiked to 23 mmol/L calcium chloride (final concentration) was prepared as positive control in the proteolytic activity and thrombin‐like assays.…”

Section: Methodsmentioning

confidence: 99%

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Single‐use technology for solvent/detergent virus inactivation of industrial plasma products

Hsieh

Mullin²,

Greenhalgh³

et al. 2016

Transfusion

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BACKGROUND Virus inactivation of plasma products is conducted using stainless‐steel vessels. Single‐use technology can offer significant benefits over stainless such as operational flexibility, reduced capital infrastructure costs, and increased efficiency by minimizing the time and validation requirements associated with hardware cleaning. This study qualifies a single‐use bag system for solvent/detergent (S/D) virus inactivation. STUDY DESIGN AND METHODS Human plasma and immunoglobulin test materials were S/D‐treated in Mobius single‐use bags using 1% tri‐n‐butyl phosphate (TnBP) with 1% Triton X‐100 or 1% Tween 80 at 31°C for 4 to 6 hours to evaluate the impact on protein quality. Volatile and nonvolatile organic leachables from low‐density polyethylene film (Pureflex film) used in 1‐L‐scale studies after exposure to S/D in phosphate‐buffered saline were identified compared to controls in glass containers. Virus inactivation studies were performed with xenotropic murine leukemia virus (XMuLV) and bovine viral diarrhea virus (BVDV) to determine the kinetics of virus inactivation, measured using infectivity assays. RESULTS S/D treatment in Mobius bags did not impact the protein content and profile of plasma and immunoglobulin, including proteolytic enzymes and thrombin generation. Cumulative leachable levels after exposure to S/D were 1.5 and 1.85 ppm when using 0.3% TnBP combined with 1% Tween 80 or 1% Triton X‐100, respectively. Efficient inactivation of both XMuLV and BVDV was observed, with differences in the rate of inactivation dependent on both virus and S/D mixture. CONCLUSION Effective S/D virus inactivation in single‐use container technology is achievable. It does not alter plasma proteins and induces minimal release of leachables.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Single‐use technology for solvent/detergent virus inactivation of industrial plasma products

Hsieh

Mullin²,

Greenhalgh³

et al. 2016

Transfusion

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“…Albumin determination used a bromocresol green solution (BCG) as a binding dye, and the albumin-BCG complex was measured as an endpoint reaction at 596/694 nm. The purity of the gamma protein was assessed by zone electrophoresis using a Hydragel 7 protein kit (Sebia, Evry, France) [ 22 ]. Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing or reducing conditions was run using a NuPAGE 4%~12% Bis-Tris Gel (Invitrogen, Carlsbad, CA, USA) and a PageRuler prestained protein ladder (Thermo).…”

Section: Methodsmentioning

confidence: 99%

Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

et al. 2016

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Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a severe disease that is caused by maternal alloantibodies generated during pregnancy or at delivery as a result of incompatibility between maternal and fetal human platelet antigens (HPAs) inherited from the father. Antibody-mediated immune suppression using anti-HPA-1a immunoglobulins is thought to be able to prevent FNAIT caused by HPA-1a. A fractionation process to prepare anti-HPA-1a immunoglobulin (Ig) G (IgG) from human plasma was therefore developed. Anti-HPA-1a plasma was obtained from volunteer mothers who underwent alloimmunization against HPA-1a during a previous pregnancy. Plasma was cryoprecipitated and the supernatant treated with caprylic acid and solvent/detergent (S/D), purified by chromatography, nanofiltered, concentrated, and sterile-filtered. The anti-HPA-1a immunoglobulin fraction was characterized for purity and safety. PAK12 and quantitative monoclonal antibody immobilization of platelet antigen (MAIPA) assays were used to detect anti-HPA-1a IgG. Hepatitis C virus (HCV) removal during nanofiltration was assessed by spiking experiments, using cell culture-derived reporter HCV and luciferase analysis. The caprylic acid treatment precipitated non-Ig proteins yielding a 90% pure Ig supernatant. S-HyperCel chromatography of the S/D-treated supernatant followed by HyperCel STAR AX provided high IgG recovery (>80%) and purity (>99.5%), and efficient IgA and IgM removal. Concentrations of complement factors C3 and C4 were < 0.5 and < 0.4 mg/dL, respectively. The final IgG could be nanofiltered on Planova 20N under conditions removing more than 3 log HCV infectivity to baseline mock infection level, and concentrated to ca. 30 g/L. Proteolytic activity and thrombin generation were low in the final fraction. The Pak12 and MAIPA assays showed good recovery of anti-HPA-1a throughout the process. Clinical-grade HPA-1a IgG can be prepared using a process compliant with current quality requirements opening perspectives for the prevention of FNAIT.

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“…Other methods under development use the concept of protein partitioning that is distinct from chromatography , including preparative electrophoresis technology using membranes of controlled porosity currently being implemented in Singapore, and aqueous two‐phase systems (ATPS) developed in Costa Rica so far for albumin and IgG .…”

Section: Fractionation Technologiesmentioning

confidence: 99%

Current status and new developments in the production of plasma derivatives

Burnouf

2016

ISBT Science Series

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Plasma fractionation is a unique biotechnology and bioengineering industry characterized by its capacity to extract a diverse range of protein therapeutics that are often the only options for treating life‐threatening pathological conditions of patients suffering from bleeding, immunological disorders, infections or trauma. Current plasma products include several coagulation factors, polyvalent and hyperimmune immunoglobulins (Igs), albumin, protease inhibitors and anticoagulants. In many pathological situations, these products are used for long‐term replacement therapy to compensate for a congenital or acquired protein deficiency. Most products are obtained by plasma fractionation technologies that derive from the cold ethanol fractionation process developed in the mid‐/late 1940s by Cohn and collaborators. Still, in the last 20 years, this core technology has undergone substantial evolutions illustrated by the integration of several viral reduction steps for all products and by the introduction of chromatographic purification steps to extract labile proteins from cryoprecipitate‐poor plasma. The most recent technological changes, intended mostly to improve recoveries and remove trace protein contaminants, concern the production process of intravenous Igs through implementing chromatographic steps and caprylic acid treatment in early stages of the ethanol fractionation process. A number of new fractionation technologies are currently under development and have not been licensed at least for the time being by major regulatory authorities. One target of these technologies is to improve the recovery of the most important plasma protein products, under conditions facilitating large‐scale implementation in emerging countries.

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Human plasma‐derived immunoglobulin G fractionated by an aqueous two‐phase system, caprylic acid precipitation, and membrane chromatography has a high purity level and is free of detectable in vitro thrombogenic activity

Abstract: Fractionating human plasma by an ATPS combined with caprylic acid and membrane chromatography resulted in an IgG preparation of high purity and free of a detectable in vitro thrombogenic risk.

Cited by 13 publications

References 32 publications

Single‐use technology for solvent/detergent virus inactivation of industrial plasma products

Single‐use technology for solvent/detergent virus inactivation of industrial plasma products

Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

Current status and new developments in the production of plasma derivatives

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