Southern blot test trial#

In addition to that, a typical assessment of result can be performed by comparison to the customer´s reference sample (as stated above). ORDERING INFORMATION: Geisinger Epic Procedure Code: LAB2319 Geisinger Epic ID: 35484. Testing results from the EOPC and WCB are compared to those of the MCB to allow the detection of any changes that may be indicative of cell line instability. Testing of the working cell banks (WCB) may also be performed at a manufacturer’s discretion, especially if there were any changes in the production process. The minimal characterization required is to assess the Master Cell Bank (MCB) and the End of Production Cell Bank (EOPC). Southern blots also confirm target identity as matching the hybridization probe to digested DNA sample compared to a molecular weight marker and/or digested reference sample. High quality and large amounts of DNA are needed. Southern blot analysis is usually a part of a cell line stability testing package, which is a key component in characterizing production cell banks and is critical in maintaining quality assurance of biologicals/ modern therapeutics derived from bacterial, yeast and mammalian cell cultures.īasically, ICH Guidelines require Southern blots (among other methods) to confirm that the structure of the genomic DNA of a cell bank is stable and unchanged and that no clonal rearrangement has occurred. Compared to other tests, southern blot is a complex technique that has multiple steps and these steps require equipment and reagents that are expensive. The next step involves transfer and UV cross-linking of the DNA fragments to a membrane and hybridization with a specific nucleic acid probe, which will identify DNA target regions of interest. Then, a certain amount of DNA is digested by a specific restriction endonuclease and cut DNA samples are separated by agarose gel electrophoresis. In this method, DNA is first extracted from the sample and quantified by UV spectrometry. Southern blot analysis is one of the oldest methods that is used to analyze DNA.

Southern blot test trial#

GLP Pre-clinical safety and clinical trial testingĪssay Development, Validation and Transfer Master, Working and End-of-production cell bank testing Retrovirus assay - reverse transcriptase enzyme activity (qPBRT) assay The probe used in the test is a small piece of DNA from the. It is a multi-step process, that begins with the electrophoresis of DNA, transfer of DNA fragments into nitrocellulose strip, and exposing those fragments to a DNA probe labeled with a radioactive or chemical tag. Viral seed qPCR adventitious agents testing The digested DNA is separated on an agarose gel and transferred to nylon membrane for hybridization. Southern blotting is a molecular technique to find target DNA sequences in a sample. ATMPs testing (Cell therapy / Gene therapy / Tissue-engineered therapy testing)Ĭell line authentication - DNA genotyping.Custom assay design, validation and transfer