Preparation and standardization of an internal reference for bovine serum albumin residue detection kit

Abstract

Objective To prepare an internal reference to investigate the stability of bovine serum albumin (BSA) residue detection kit. Methods BSA residues were detected in 3 working liquids (virus diluent, virus soaking solution and cell supernatant) during the production process of a live attenuated Japanese encephalitis vaccine. The liquid with appropriate BSA content was diluted by vaccine freeze-drying protective agent to prepare the internal reference. The reference was standardized 3 times at different dates by 2 laboratory workers using 3 batch kits, then the assignment range was determined. The detection results from different batch kits and different workers were compared by ANOVA and t-test, respectively. In the following 8 months, the internal reference was used 20 times to investigate the stability of the detection kit. Results According to the BSA detection results, virus soaking solution was chosen and 3-fold diluted by freeze-drying protective agent to prepare the internal reference. The mean value (Mean), standard deviation (SD) and coefficient of variation (CV) of 108 detection values were 33.46 ng/ml, 2.15 ng/ml, and 6.40%, respectively. The assignment range of the reference was determined as 27.01-39.91 ng/ml.There was significant difference among the detection results from different batch kits (F=11.497, P 0.05). The internal reference was detected 20 times using 5 batch kits, and BSA contents were in the range of 30.00-36.63 ng/ml, with Mean of 33.14 ng/ml, SD of 1.73 ng/ml, CV of 5.20%. There was no significant difference among the results from different batch kits (F=0.997, P>0.05). Conclusion The internal reference can be used to investigate the stability of BSA residue detection kit. Key words: Serum albumin, bovine; Reference standards; Reagent kits; Japanese encephalitis vaccines