Abstract
BackgroundThe quality and safety of advanced therapy products must be maintained throughout their production and quality control cycle to ensure their final use in patients. We validated the cell count method according to the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use and European Pharmacopoeia, considering the tests’ accuracy, precision, repeatability, linearity and range.MethodsAs the cell count is a potency test, we checked accuracy, precision, and linearity, according to ICH Q2. Briefly our experimental approach was first to evaluate the accuracy of Fast Read 102® compared to the Bürker chamber. Once the accuracy of the alternative method was demonstrated, we checked the precision and linearity test only using Fast Read 102®. The data were statistically analyzed by average, standard deviation and coefficient of variation percentages inter and intra operator.ResultsAll the tests performed met the established acceptance criteria of a coefficient of variation of less than ten percent. For the cell count, the precision reached by each operator had a coefficient of variation of less than ten percent (total cells) and under five percent (viable cells). The best range of dilution, to obtain a slope line value very similar to 1, was between 1:8 and 1:128.ConclusionsOur data demonstrated that the Fast Read 102® count method is accurate, precise and ensures the linearity of the results obtained in a range of cell dilution. Under our standard method procedures, this assay may thus be considered a good quality control method for the cell count as a batch release quality control test. Moreover, the Fast Read 102® chamber is a plastic, disposable device that allows a number of samples to be counted in the same chamber. Last but not least, it overcomes the problem of chamber washing after use and so allows a cell count in a clean environment such as that in a Cell Factory. In a good manufacturing practice setting the disposable cell counting devices will allow a single use of the count chamber they can then be thrown away, thus avoiding the waste disposal of vital dye (e.g. Trypan Blue) or lysing solution (e.g. Tuerk solution).
Highlights
The quality and safety of advanced therapy products must be maintained throughout their production and quality control cycle to ensure their final use in patients
According to the International Conference on Harmonization (ICH) International conference on harmonization Q2 (Q2) [9]: “accuracy expresses the closeness of agreement between the value which is accepted as either a conventional true value or as an accepted reference value and the value found; precision of an analytical procedure expresses the closeness of agreement between a series of measurements obtained from the multiple sampling of the same homogeneous sample under the prescribed conditions; repeatability expresses the precision under the same operating conditions over a short interval of time; linearity of an analytical procedure is its ability to obtain test results which are directly proportional to the concentration of an analyte in the same sample”
Two cell subpopulations were chosen for the validation procedure: mononuclear cells (MNCs), as a prototype of lymphocytes, and mesenchymal stem cells (MSCs), both cell therapy products (CTP) that we will produce for immunotherapy and regenerative medicine
Summary
The quality and safety of advanced therapy products must be maintained throughout their production and quality control cycle to ensure their final use in patients. Some automatic counters are marketed, for GMP settings, their associated software should comply with 21 CFR Part 11 [11,12,13,14] On these bases, our primary aim was to validate a disposable cell count method in GMP conditions, to be included in the Validation Master Plan (VMP) to be submitted to the Regulators, to obtain accreditation of our Cell Factory to produce CTPs. We decided to use a manual cell count and validate, according to GMP rules, a disposable device, Fast Read 102 W, already used in P3 laboratory by our group
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