DNA Sizing and Barcoding: Alternatives to Isoenzyme Analysis
Cell line identity testing is a critical regulatory requirement for recombinant cell lines in order to confirm the cell line’s species of origin, as well as to assist in detecting contamination from other cell lines.
Biochemical analysis of isoenzyme polymorphism has historically been considered the standard test for mammalian and insect cell line identification; however, the methodology suffers several limitations, including:
- Limited species coverage
- Lack of sufficient sensitivity
- Difficulty in data interpretation, particularly when trying to differentiate closely related species
- Test kits provided by a single supplier
- Recent shortage of reagent supply
To overcome these limitations and challenges, Eurofins Lancaster Laboratories has developed DNA Sizing and Barcoding Methods for species identification. These methods utilize the mitochondrial genes Cytochrome Oxidase 1 (CO1) and Cytochrome B (Cyt B) as targets for the molecular identification of cell lines commonly used in biopharmaceutical production.
Why Choose Eurofins Lancaster Laboratories?
Our comprehensive cell line characterization services, along with our cell banking capabilities, provide you with a single-source solution for all of your cell line needs.
CO1 DNA Sizing and Barcoding Method
- For CO1/Cyt B Sizing, genomic DNA is isolated from the cells being tested and amplified using a set of species specific primers.
- Species identification and detection of cross contamination can be made based upon the presence/absence of PCR amplification by given sets of primers and the size of the PCR amplicon. Cell lines of cow, mouse, dog, rat, monkey, hamster, cat, human and pig origin can be tested using CO1/Cyt B Sizing.
- Identification of cell lines derived from a wider variety of animal and insect species can be made by CO1 DNA Barcoding. The comparison between the determined CO1 sequence of tested cell lines and the confirmed species specific sequences deposited in the Consortium for the Barcode of Life database allows for unambiguous genetic identification.
- The Sizing and Barcoding Methods are based on a more robust technology than that of the isoenzyme assay and can therefore provide more accurate speciation. They can also be easily implemented in a quality control environment.
- The PCR amplification and DNA sequencing techniques used for the Sizing and Barcoding Methods decrease dependency on a single supplier for test kits and reagents.
- The Sizing and Barcoding Methods are more sensitive with regard to the detection of cell line cross contamination than isoenzyme analysis.
Please contact us for a technical discussion prior to sample submission.
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