Control of Large-Scale Plasma Thawing for Recovery of
Cryoprecipitate Factor VIII

Foster, Peter R.; Dickson, Alan J.; McQuillan, Thomas A.; Dickson, Ida H.; Keddie, Samuel; Watt, John G.

doi:10.1159/000460876

Cited by 6 publications

(8 citation statements)

References 17 publications

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“…Although the production of cryoprecipitate is a simple procedure, it has two major disadvantages, the recovery of FVIII is low about 40% of that in normal plasma, and the purity is poor (2,3). To overcome these disadvantages, the cryoprecipitate is further purified using various techniques such as adsorption (4), gel filtration (5) and ion exchange chromatography (6,7,8).…”

Section: Indian Journal Of Clinical Biochemistry 2003mentioning

confidence: 99%

Separation of antihemophilic factor VII from human plasma by column chromatography

Baikar¹,

Kamath²,

Vishwanathan³

et al. 2003

Indian J Clin Biochem

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TMAE-Fractogel 650 (M) (Trimethylaminoethyl) is an ion exchange medium can be used to capture factor Vlll (F VIII) directly from plasma. Previous reports have focused on the use of DEAE-Fractoge1650 (M) (Dimethylaminoethyl) ion exchange medium to capture F Vlll from cryoprecipitate and plasma. Our main objectives were (I) to standardise the purification of FVIII from human plasma by column chromatographic technique. (11) to study the recovery of FVIII activity in purified fraction at 18 -20~ process condition. (Ill) to study the effect of virucidal step on recovery of FVIII activity and (IV) to study the effect of lyophilisation on FVIII activity. In this report, Citrate Phosphate Dextrose (CPD) plasma was batch stirred with dry DEAE-Sephadex A50, filtered, diluted, loaded on to a column packed with TMAE-Fractogel and chromatographed. Most of the unwanted proteins flowed through the gel unadsorbed. Bound F VIII was eluted by increasing the ionic strength of the buffer. This purification step gave an overall 80% recovery from the plasma with a specific activity of 0.97 IU FVlll/mg protein. The purified F VIII fraction was made virus safe by employing the virucidal technique developed by New York Blood Centre (NYBC). There was 48.43% loss of FVIII activity in Virus inactivation treatment and the loss of FVIII activity in lyophilisation was 8.45% which is acceptable. This method of purification gave a higher yield of FVIII than cryoprecipitation, and is a promising alternative method to cryoprecipitation of F VIII.

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Section: Indian Journal Of Clinical Biochemistry 2003mentioning

confidence: 99%

Separation of antihemophilic factor VII from human plasma by column chromatography

Baikar¹,

Kamath²,

Vishwanathan³

et al. 2003

Indian J Clin Biochem

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“…The blood group of the donations was not taken into account in the construction of plasma pools for fractionation. Factor VIII concentrate was pre pared (table I) from pools of plasma (range from about 300-700 kg) in a manner similar to that previously described [5] using the contin uous procedure for plasma thawing. The resultant cryoprecipitate was processed (table I) [5] using the low-shear agitator (Vibromixer, model E2; Chemap, Abingdon, UK) but the final pH adjustment (step 12, table I) was to pH 6.8 using hydrochloric acid rather than citric acid.…”

Section: Preparation Of Fviii Concentratementioning

confidence: 99%

“…Factor VIII concentrate was pre pared (table I) from pools of plasma (range from about 300-700 kg) in a manner similar to that previously described [5] using the contin uous procedure for plasma thawing. The resultant cryoprecipitate was processed (table I) [5] using the low-shear agitator (Vibromixer, model E2; Chemap, Abingdon, UK) but the final pH adjustment (step 12, table I) was to pH 6.8 using hydrochloric acid rather than citric acid. 1 Results from this study have been presented in preliminary form at meetings of the British Society of Haematology [1], The World Federation of Haemophilia [2] and the International Society of Thrombosis and Haemostasis [3].…”

Section: Preparation Of Fviii Concentratementioning

confidence: 99%

“…FPA measurements were carried out by radioimmunoassay [11] using the reagents and method supplied by IMCO (Sweden) with minor modification. Measurements of total protein and the recon stitution time of freeze-dried preparations were carried out as pre viously described [5],…”

Section: Clinical Evaluation Of a Selected Formulationmentioning

confidence: 99%

“…product formulation, 0.22pm membrane filtration and aseptic dispensing) prior to freeze-drying. Although re design of the cryoprecipitation procedure has given a substantial yield improvement [2, 5,6], we have contin ued this project by examining the VIIIiC loss over product finishing and it is the results of this study which are reported here.…”

Section: Introductionmentioning

confidence: 99%

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Studies on the Stability of VIII:C during the Manufacture of a Factor VIII Concentrate for Clinical Use

Foster¹,

Dickson²,

McQuillan³

et al. 1988

Vox Sanguinis

Self Cite

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The stability of VIII:C was investigated by monitoring samples taken at different points from a routine process for the manufacture of factor VIII concentrate and by examining the stabilising influence of a number of product formulations. Loss of VIII:C over process-finishing procedures (formulation, 0.22 µm filtration, dispensing) was associated with a citrate-induced inactivation which could be prevented by controlling the ionised calcium concentration of the solution. These results were obtained using a one-stage clotting assay but were not observed using a two-stage assay. No evidence for activation was found in vitro (e.g. by FPA generation and VIII:C stability) and the yield increase suggested by the one-stage assay was supported by results from a controlled clinical evaluation.

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Direct Capture of Plasma Factor VIII:C by Ion Exchange Chromatography

Teh¹,

Froger²

1994

Vox Sanguinis

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An ion exchange medium, DEAE-Fractogel 650M, can be used to capture factor VIII (FVIII) directly from plasma. Previous reports have focussed on the use of this medium to capture FVIII from cryoprecipitate. In this report, citrate phosphate dextrose plasma was batch-stirred with DEAE-Sephadex A50, filtered, diluted, loaded onto a column packed with DEAE-Fractogel TSK 650M, and chromatographed. Most of the unwanted proteins flowed through the gel unadsorbed. Bound FVIII was eluted by increasing the ionic strength of the buffer. A citrate- based buffer gave an overall FVIII:C yield of 670 IU/kg plasma with a specific activity of 0.68 IU FVIII:C/mg protein. This process gave a higher yield of FVIIEC but significantly more prothrombin complex factors than cryoprecip- itation. This method is a promising alternative to cryoprecipitation of FVIII.

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Control of Large-Scale Plasma Thawing for Recovery of Cryoprecipitate Factor VIII

Cited by 6 publications

References 17 publications

Separation of antihemophilic factor VII from human plasma by column chromatography

Separation of antihemophilic factor VII from human plasma by column chromatography

Studies on the Stability of VIII:C during the Manufacture of a Factor VIII Concentrate for Clinical Use

Direct Capture of Plasma Factor VIII:C by Ion Exchange Chromatography

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