Progress in large‐scale purification of factor VIII/von Willebrand factor concentrates using ion‐exchange chromatography

Mori, Filippo; Nardini, Ilaria; Rossi, Paola; Nardini, Claudia; Farina, Claudio

doi:10.1111/j.1423-0410.2008.01096.x

Cited by 11 publications

(12 citation statements)

References 29 publications

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“…As shown in Figures 2 and 3B (fractions 5 and 6) and Tables 2, 3 and 7, IaIp (heavy chains 1, 2 and 3 and light chain bikunin) are the main impurities in both F VIII concentrates, Octanate and Haemoctin. Unfortunately, the quantitative analysis of these proteins is still not a part of routine quality control of F VIII concentrates [11, 23]. Lim et al recently developed a competitive ELISA for determination of IaIp, and we used this test for quantitative determination of these proteins in F VIII concentrates (cf.…”

Section: Discussionmentioning

confidence: 99%

Proteomic characterization of plasma‐derived clotting factor VIII–von Willebrand factor concentrates

et al. 2009

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Proteomic methods were used to identify the levels of impurities in three commercial plasmaderived clotting factor VIII-von Willebrand factor (FVIII/VWF) concentrates. In all three concentrates, significant amounts of other plasma proteins were found. In Octanate and Haemoctin, two concentrates developed in the 1990s, the major impurities identified were interalpha inhibitor proteins, fibrinogen and fibronectin. These two concentrates were also found to contain additional components such as clotting factor II (prothrombin) that are known activators of FVIII. In Wilate, a recently developed FVIII/VWF concentrate, the amount of these impurities was significantly reduced. Batch-to-batch variations and differences between three investigated products were detected using iTRAQ, an isotope labeling technique for comparative mass spectrometry, demonstrating the potential value of this technique for quality control analysis. The importance of thorough proteomic investigations of therapeutic F VIII/VWF preparations from human plasma is also discussed.

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Section: Discussionmentioning

confidence: 99%

Proteomic characterization of plasma‐derived clotting factor VIII–von Willebrand factor concentrates

et al. 2009

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“…It has the advantage of high loading abilities. This chromatographic mode was reported for the isolation of vitamin K‐dependent clotting factors II, VII, IX, and X and clotting inhibitors protein C and protein S, as well as interalpha inhibitor proteins, von Willebrand factor [67–69].…”

Section: Sample Preparationmentioning

confidence: 99%

Proteomics applied to transfusion plasma: the beginning of the story

et al. 2013

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'Safe blood' is and has always been the major concern in transfusion medicine. Plasma can undergo virus inactivation treatments based on physicochemical, photochemical or thermal methodologies for pathogen inactivation. The validation of these treatments is essentially based on clottability assays and clotting factors' titration; however, their impact on plasma proteins at the molecular level has not yet been evaluated. Proteomics appears as particularly adapted to identify, to localize and, consequently, to correlate these modifications to the biological activity change. At the crossroads of biology and analytical sciences, proteomics is the large-scale study of proteins in tissues, physiological fluids or cells at a given moment and in a precise environment. The proteomic strategy is based on a set of methodologies involving separative techniques like mono- and bidimensional gel electrophoresis and chromatography, analytical techniques, especially mass spectrometry, and bioinformatics. Even if plasma has been extensively studied since the very beginning of proteomics, its application to transfusion medicine has just begun. In the first part of this review, we present the principles of proteomics analysis. Then, we propose a state of the art of proteomics applied to plasma analysis. Finally, the use of proteomics for the evaluation of the impact of storage conditions and pathogen inactivation treatments applied to transfusion plasma and for the evaluation of therapeutic protein fractionated is discussed.

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“…This coagulation factor is an essential glycoprotein for the clotting process. It circulates in the bloodstream complexed with von Willebrand factor, which acts as a transporter [ 3 ] and prevents the degradation of FVIII by the action of serine proteases, including activated Protein C and activated FX [ 4 ]. FVIII is a high molecular mass (300 kDa) and labile protein present in very low concentrations in plasma (150 ng mL −1 ) [ 4 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

“…Anion exchange resins are widely used in the plasma fractionation processes [ 3 , 12 , 13 , 14 , 15 ] due to their high capacity, good yields and low cost compared to other chromatographic techniques. Upon loading whole plasma directly onto an anion exchange column, without the cryoprecipitation step, FVIII and PCC are strongly adsorbed and cannot be eluted separately using NaCl gradient [ 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

Non-Cryoprecipitation Separation of Coagulation FVIII and Prothrombin Complex Proteins by Pseudoaffinity Calcium Elution Chromatography Using Anion Exchange Resin

Feliciano

Arimori²,

Nakao³

et al. 2022

Pharmaceuticals

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Hemophilia A is treated with human plasma coagulation factor VIII (FVIII) replacement therapy and Hemophilia B with coagulation factor IX, which is purified from prothrombin complex concentrate (PCC). In this paper we evaluated the separation of FVIII and PCC by directly loading raw thawed plasma to an anion exchange resin (AEX). Under this relatively high ionic strength, most of the plasma proteins such as albumin, immunoglobulins and others were not adsorbed. Five resins commonly used in protein purification (plasma fractionation) were tested. With all resins, PCC was eluted by pseudoaffinity in a calcium gradient step. Afterwards, FVIII could be recovered with a good yield and high purification factor in the salt gradient step with 400–500 mM NaCl. Using ANX Sepharose FF and Q Sepharose FF, the CaCl2 elution step was introduced after the intermediate wash with 200 mM NaCl, whereas using DEAE Sepharose FF, Fractogel EMD TMAE and Fractogel EMD DEAD, PCC eluted after the wash of the unbound proteins. Our results indicate that three important fractions: (1) albumin, immunoglobulin etc.; (2) PCC; and (3) FVIII can be separated in one chromatographic AEX column and the delicate and troublesome cryoprecipitation can be eliminated, making the purification of blood products faster and cheaper.

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Progress in large‐scale purification of factor VIII/von Willebrand factor concentrates using ion‐exchange chromatography

Abstract: Several highly purified products, suitable for haemophilia A and von Willebrand disease management, can be obtained, through the same chromatographic process, on an industrial scale.

Cited by 11 publications

References 29 publications

Proteomic characterization of plasma‐derived clotting factor VIII–von Willebrand factor concentrates

Proteomic characterization of plasma‐derived clotting factor VIII–von Willebrand factor concentrates

Proteomics applied to transfusion plasma: the beginning of the story

Non-Cryoprecipitation Separation of Coagulation FVIII and Prothrombin Complex Proteins by Pseudoaffinity Calcium Elution Chromatography Using Anion Exchange Resin

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