DETERMINATION OF COAGULATION FACTOR VIII ACTIVITY IN CRYO-FREE PLASMA SAMPLES: AN ANALYSIS OF RAW MATERIAL SUITABILITY FOR INDUSTRIAL USE

Abstract

According to the Brazilian Pharmacopoeia, Human Plasma for Fractionation, raw material of the Blood-Derivative Drugs Industry, must present a concentration of Coagulation Factor VIII (FVIII) greater than or equal to 0.7 IU/ mL. The Crio-Free Plasma (CFP) is not captured by the Blood-Derivative Drugs Industry due to its low FVIII content, hence its production is mostly discarded, generating high financial costs for brazilian blood centers. This study aims to evaluate the adequacy of Cryo-Free Plasma to the parameters required by the Brazilian Pharmacopeia regarding the concentration of FVIII for Human Plasma for Fractionation, as well as analyzing possible critical points in the process of obtaining this plasma that interfere with the final FVIII content of the product. Two pools of 10 units of Fresh Frozen Plasma (FFP) and CFP each were prepared. Aliquots of these pools were mixed in different proportions and the activity of FVIII was dosed in duplicate. The activity of FVIII was also measured in triplicate in CFP pool samples that had been frozen in 8 and 24 hours after collection. The sample containing only CFP revealed a recovery of FVIII between 42-46%. By mixing CFP and FFP from the 1:1 ratio, the results indicated an activity of FVIII above 70%, and this activity was higher with the increase in the proportion of FFP. The mean percentage of FVIII activity present in CFP samples frozen after 24 hours of the plasma collection was 15.1% (SD ± 0.007), while CFP samples frozen within 8 hours obtained an average activity of 43.1% (SD ± 0.027). Although CFP alone presents an amount of FVIII below what is recommended in the Brazilian pharmacopoeia for industrial fractionation, this blood product could still be used by the industry to obtain other plasma proteins. In order to overcome the FVIII deficiency, a pool with FFP in determined proportions can be made, obtaining a acceptable final concentration of FVIII. However, the freezing time after its collection in blood centers remains an important factor to be monitored as a quality criterion for the maintenance of Factor VIII content in plasma. The discussion about the regulatory aspects of CFP utilization by the Blood-Derivative Drugs industry is important, as it contributes to a better exploitation of raw material and significant cost savings for the brazilian blood centers, that would no longer have to discard large amounts of CFP per year. CFP could be used as raw material if it is mixed in a minimum ratio of 1:1 to the FFP and is frozen within 8 hours after its collection, preserving the activity of the FVIII. Further studies are needed to assess whether these results can be validated on an industrial scale.