Abstract
Background aims
Cell therapy products (CTP) typically require full sterility, endotoxin and Mycoplasma testing before product release. Often this is not feasible with fresh cells, and
sponsors may rely on rapid microbiological methods (RMM). RMM must be qualified in-house
using the sponsor's facilities, equipment, consumables, cells and matrices to meet
regulatory approval. Herein, we present a cost-effective strategy to conduct an in-house
abbreviated qualification of a commercially available RMM kit to meet Health Canada
regulatory requirements.
Methods
We performed an abbreviated qualification using a polymerase chain reaction (PCR)-based
Mycoplasma testing method involving assay sensitivity and ruggedness, based on an experimental
plan that was pre-approved by Health Canada. Briefly, investigational CTPs were tested
in-house using a PCR-based Mycoplasma detection kit. Assay sensitivity was determined using a 10-fold dilution series of
genomic DNA of only two Mycoplasma species, Mycoplasma arginini and Mycoplasma hominis in the absence of CTP-matrix as the kit had been previously validated against nine
species. Matrix interference was measured by testing independent CTP samples. Testing
by different operators on different days measured ruggedness.
Results
The RMM Mycoplasma qualification exceeded sensitivity (4 genome copies per reaction for M. arginini and 0.12 genome copies per reaction for M. hominis) and met ruggedness requirements without matrix interference, as required by the
Pharmacopoeial guidelines (Ph. Eur. 2.6.7 and USP <1223>).
Discussion
Our approach represents a minimal qualification that can be performed by an academic
institution while ensuring regulatory compliance for implementing RMM testing for
in-process and product-release testing of CTPs.
Key Words
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Article info
Publication history
Published online: October 20, 2017
Accepted:
September 3,
2017
Received:
May 18,
2017
Identification
Copyright
© 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
