Isolation of the factor VIII–von Willebrand factor complex directly from plasma by gel filtration

Kaersgaard, P.; Barington, Karina A.

doi:10.1016/s0378-4347(98)00250-3

Cited by 11 publications

(6 citation statements)

References 27 publications

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“…Size exclusion chromatography (SEC) is one efficient method for the purification of proteins derived from human plasma (2). SEC is primarily suited for the final phase in a separation process, the so-called polishing step (3).…”

Section: Introductionmentioning

confidence: 99%

Continuous Removal of Protein Aggregates by Annular Chromatography

Buchacher

Iberer

Jungbauer³

et al. 2001

Biotechnol. Prog.

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The removal of polymeric proteins from their monomers is a frequently encountered separation task, especially in the polishing step of therapeutic proteins. Continuous separation of protein polymers from monomers by annular chromatography using size exclusion chromatography has been studied regarding the resolution, recovery, fouling, and productivity and has been compared to conventional chromatography. An IgG preparation rich in aggregates was used as a model protein mixture. Under conditions that maximized the throughput, the polymers could be separated from the monomers, but baseline separation could not be achieved. Baseline separation was also not possible in batch mode using equivalent conditions, which was also confirmed by computer simulation. For separation of the aggregates from the product the entire available separation space (360 degrees ) was indispensable. Therefore only cyclic, discontinuous regeneration could be carried out. Loading was identified as a critical step, since the concentrated protein solution evaded into the headspace instead of migrating into the gel where viscous fingering often occurs in conventional chromatography. The productivity of annular chromatography was two times higher than that of the conventional batch chromatography, and the buffer consumption was reduced to half the conventional value. These two benefits are especially important for protein separation processes that suffer from low loadability, such as size exclusion chromatography. We have demonstrated that size exclusion can be performed on an industrial scale when it is run continuously with the aid of a pressurized annular chromatograph.

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Section: Introductionmentioning

confidence: 99%

Continuous Removal of Protein Aggregates by Annular Chromatography

Buchacher

Iberer

Jungbauer³

et al. 2001

Biotechnol. Prog.

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“…Several reports of plasma protein purification by size exclusion chromatography are available; but none were performed with the type of column and buffer system used in this work. By comparing the chromatographic profile of the gel filtration of human plasma presented in Figure 1 with other works, it is possible to observe that the chromatographic profile can vary widely for the same sample at the expense the column used [41,42].…”

Section: Discussionmentioning

confidence: 90%

“…In this work, the buffer composed of 3.2% Sodium Citrate pH 8.0 was used in all steps of protein purification. The majority of the previously solutions used for plasma purification were buffered with Tris-HCl [23,41,42]. The choice of 3.2% Sodium Citrate was fundamental to better separation of plasma proteins since this buffer prevents the activation of the coagulation cascade, by sequestering calcium, and it is a good buffering at a pH close to physiological, favoring proteins to maintain the conformation found in plasma.…”

Section: Discussionmentioning

confidence: 99%

“…Serotransferrin is associated with Fe 3+ uptake, distribution and control of its quantity [47]. Antithrombin-III is an endogenous anticoagulant capable of inhibit key-factors of the coagulation cascade such as thrombin and FXa, thus, having a crucial role in homeostasis [42]. About 25% of the total NSIAF from peak 1 and peak 3 (Figures 2A and 2C) was decurrently from serotransferrin and antithrombin-III, respectively.…”

Section: Discussionmentioning

confidence: 99%

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A Method for Purifying Native Transthyretin from Human Plasma

Santana¹,

Carvalho²,

Zanchin³

et al. 2015

Biochem Pharmacol (Los Angel)

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Transthyretin is a homotetrameric thyroid-hormone-transporting protein that binds to the retinol binding protein thus being involved in metabolism, growth, fertility, homeostasis of the cardiovascular and central nervous system, cell differentiation, reproduction, development and maintenance the cognitive processes during aging. Currently, there are several methodologies for natively purifying TTR from plasma, serum, tears, and amyloid fibrils; however, these procedures are laborious. Herein, a low-cost and simple protocol to purify TTR from human plasma is described. It involves the separation of plasma proteins by size exclusion and DEAE chromatography. The homogeneity was assessed by SDS-PAGE and by tandem mass spectrometry using an Orbitrap-XL (Thermo, San Jose-CA).

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“…The gel filtration resin Sepharose 4 Fast Flow is a successful method to separate FVIII/vWF from smaller molecular mass proteins (Kaersgaard and Barington 1998). Therefore it could be a very convenient method combined with the anion exchange for FVIII purification.…”

Section: Introductionmentioning

confidence: 99%

Purification of coagulation factor VIII using chromatographic methods. Direct chromatography of plasma in anion exchange resins

et al. 2010

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Coagulation factor VIII (FVIII) concentrates are used in the treatment of patients with Hemophilia A. Human FVIII was purified directly from plasma using anion exchange chromatography followed by gel filtration. Three Q-Sepharose resins were tested, resulting in 40% recovery of FVIII activity using Q-Sepharose XL resin, about 80% using Q-Sepharose Fast Flow and 70% using the Q-Sepharose Big Beads. The vitamin K-dependent coagulation factors co-eluted with FVIII from the anion exchange columns. In the second step of purification, when Sepharose 6FF was used, 70% of FVIII activity was recovered free from vitamin K-dependent factors.

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Isolation of the factor VIII–von Willebrand factor complex directly from plasma by gel filtration

Cited by 11 publications

References 27 publications

Continuous Removal of Protein Aggregates by Annular Chromatography

Continuous Removal of Protein Aggregates by Annular Chromatography

A Method for Purifying Native Transthyretin from Human Plasma

Purification of coagulation factor VIII using chromatographic methods. Direct chromatography of plasma in anion exchange resins

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