Method Validation of Cell Proliferation Assay for Filgrastim Potency Testing

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Sompong Sapsutthipas
Jiradej Patchim
Saywarul Jadoonkittinan
Supaporn Phumiamorn

Abstract

Nowadays, biosimilar filgrastim is being to produce in domestic, quality control of filgrastim products is therefore necessary. The strength examination by using a cell proliferation method is common used in many countries. The study aimed to measure the accuracy of the methods in various parameters which include: specificity, accuracy, precision, linearity, and robustness. The results showed that this method revealed specificity to only filgrastim standard and samples. They were able to trigger the proliferation of adapted M-NSF-60 cells when compared with the formulation buffer. For the accuracy, the relative potency of filgrastim standard at 50, 100, and 150% was analyzed and showed that the %recovery were within the acceptance criteria from 80 to 125%. The precision of the method was less than 25% of GCV. Linearity and range exhibited the r2 = 0.9979 in the suitable range at 50-200% of relative potency. The robustness of the method was tested by modifying the passage numbers of cells and the incubation time. The robustness indicated that the 4 to 20 passage numbers of cells was able to use and incubation time by 27 and 29 hours, whereas %GCV was less than 25% at 95% confidence level (p = 0.11). All of the results confirmed that the cell proliferation method had the specificity, accuracy, precision, and
robustness. It is clear that this method is suitable to use as a standard method for the potency test of filgrastim products.

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