Cryoprecipitate production: the use of additives to enhance the yield

Abstract

Cryoprecipitate is still widely used to treat hemophilia A in developing countries. However, the yield of factor VIII is relatively low averaging, i.e. only 50%. We have attempted to enhance the yield by adding sodium citrate to the plasma following the method of Shanbrom and Owens (Blood 98, 2001, 60a). Fresh-frozen plasma (FFP) units were processed either as control plasma or after the addition of 10% sodium citrate. Cryoprecipitate was produced from both. After resuspension, calcium chloride was added to the citrated cryoprecipitate to correct for excess citrate prior to testing. The levels of FVIII and fibrinogen were determined in both preparations. The citrated cryoprecipitates had varying yields of fibrinogen and FVIII in the cryoprecipitate. The FVIII levels varied from 34% to 215% recovery. Fibrinogen ranged from 55.5% to 121.4%. We found that the addition of increasing amounts of CaCl2 to normal plasma raised the FVIII values from 1.0 to 4 U/ml. To determine the possibility of assay influence we added different quantities of CaCl2 to control plasma and measured the FVIII and activated partial thromboplastin time levels. Addition of citrate to plasma resulted in an increased total amount of cryoprecipitate much of which was citrate. Assays showed considerable ranges in the quantity of FVIII and fibrinogen. Activation of FVIII can be caused by addition of excess calcium.

Cryoprecipitate prepared from plasma frozen within 24 hours after phlebotomy contains acceptable levels of fibrinogen and VIIIC

“…Numerous previous studies have demonstrated that these minimum levels are achievable when the starting product is FFP, 14‐18 although reduction in the recovery of some factors occurred when certain pathogen inactivation techniques were attempted 17,18 . To improve the yield of VIIIC, collecting the plasma destined for cryo production in heparin instead of citrate has been shown to be efficacious, 19 and using large quantities of sodium citrate has likewise been demonstrated to increase the recovery of both VIIIC and fibrinogen, 16,20 although the results seem to be variable 16 . As blood centers in North America are now avoiding the manufacture of plasma products from female donors due to the risk of TRALI from HLA antibodies, sufficient inventories of FFP may become jeopardized.…”

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

“…The recovery of about 50% FVIII activity is comparable with the obtained from cryoprecipitates (Yousef et al 2006). All plasma-derived FVIII concentrates produced currently are purified from cryoprecipitate, which isolation requires refrigerated continuous centrifuges (Burnouf 2007).…”

Novel cryoprecipitate for wound healing and skin grafts in rats

“…Cryoprecipitate is prepared by slowly thawing fresh frozen plasma resulting in the formation of precipitate that can be resuspended in plasma and stored. Attempts have been made to improve the yield of factor VIII and fibrinogen to meet the requirements of the American Association of Blood Banks for blood products (17)(18)(19)(20).…”