Study On The Impact Of Protein Contents In Human Coagulation Factor Products On The Results Of Potency Detection

Human coagulation factor products, such as human coagulation factor Ⅷ, human prothrombin complex and human coagulation factor IX, are prepared after separation, purification and virus inactivation using the plasma of healthy humans as raw materials, and are mainly used in clinical practice for various coagulation factor deficiencies. The coagulation factor titer of these human coagulation factor products is an indicator of the efficacy of drugs, and the detection of the potency of the coagulation factor is a key detection item in drug quality detection. Manufacturers of blood products and detection agencies at all levels generally use full-automated coagulation instruments to detect the the potency of the factor according to the instruments, while the comparison of results using different detection methods show that the detection results of full-automatic coagulation instruments are different from those of one step method from the Chinese Pharmacopoeia in potency of some factors. The differences may be caused by different protein contents in the diluent used in sample dilution, so we herein mainly study the impact of protein contents in human coagulation factor products on the results of potency detection. Human coagulation factor products are the high-purity concentrates of coagulation factors in the blood, and the samples should be diluted in the detection of potency. However, the difference in protein contents in the diluent affects the detection results. There are few domestic manufacturers of human coagulation factor products and few studies on the detection methods for the potency of factor products. The studies mainly focus on the factor potency’s modes of operation, experimental conditions and the impact of accessories. In this subject, search of the reasons for the difference in the results of different detection methods during the actual work was set as the starting point, and the impact of the properties of different factor products and the protein contents of different dilutions on the detection results of potency was established. And finally, the dilution method suitable for the detection of factor potency was determined by means of method validation, thereby improving methods for the detection of potency of factor-based blood products. The full-automated coagulation instrument was designed primarily for the detection of clinical plasma samples, and therefore, when full-automated coagulation method was established for the detection of tier in human coagulation factor products, the impact of the properties of the products themselves on the detection results should be eliminated, and method validation should be performed on the established detection methods. In this study, the full-automated coagulation instrument was established for the detection of potency via the study on the impact of protein contents on the detection of factor potency, eliminating the impact of the properties of products themselves and detecting the factor potency accurately. The establishment of the method could not only accurately control the quality of human coagulation factor blood products to ensure that manufacturers could continuously and stably produce drugs of the same quality, but also reduce the risk of human coagulation factor products in clinical use.In this subject, the differences in the detection results during the work process were used as the starting point, and the impact of protein contents in the detection of factor potency was studied. Comparison of detection with the same type of products in foreign countries was performed, and lastly, the full-automated coagulation factor instrument for the detection of potency was established and validated.The concrete research contents are as follows:(1) For the same batch of human coagulation factor Ⅷ and human prothrombin complex samples, different diluents were selected for potency detection using the automated coagulation analysis method to determine the differences in the results of different diluents. Besides, the continuously produced human coagulation factor Ⅷ and human prothrombin complex were subjected to titer detection using different diluents to determine the differences in the results of different diluents.(2)Human coagulation factor Ⅷ and human prothrombin complex of the same batch were subjected to dilution with diluents containing different concentrations of proteins or plasma deficient in coagulation factors to determine the impact of protein concentrations on the potency detection of factors, and the same test was performed on 2 batches of the same products produced overseas to determine the dilution method suitable for the potency detection of factors.(3)Potency detection of human coagulation factor Ⅷ and human prothrombin complex using the full-automatic coagulation instrument was established and method validation was performed, and dilution method and diluents suitable for titer detection were selected. The linear relationship, accuracy, precision, intra-day precision and specificity, etc. of the method were subjected to method validation to confirm whether the established method is suitable for the potency detection of human coagulation factor products.