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Gene Therapy Product Testing and Research Services
The TTDSL performs these assays at an SOP-driven level with QA review to align with current FDA guidance for Phase I/II studies and the needs of the Sponsor. The TTDSL holds a Type 5 Master File (BB-MF) that may be used as a cross-reference in support of IND submissions. The following assays are offered:
- In Vitro Immortilization (IVIM) assay to assess genotoxicity of integrating viral vectors (based on Modlich et al, PMCID: PMC2835038)
- Lentiviral vector physical titer assay by p24 or viral RNA
- Lentiviral/retroviral vector infectious titer determination using HT1080 cells and qPCR analysis of integrated vector
- Lentiviral vector toxicity of human CD34+ cells with optional CFU assay or qPCR copy number determination
- Endotoxin by the LAL method
- Mycoplasma by PCR (Roche MycoTOOL or Agilent MycoSensor)
Short on personnel or time? We can also assist in developing customized testing assays. The TTDSL has a broad expertise in qPCR, ELISA, protein HPLC, cell culture, and flow cytometry-based assay development. Importantly, management has several years of experience in gene therapy product testing and manufacturing. Contact us for more information.
Fanconi Anemia Complementation Analysis
Fanconi anemia (FA) is a rare genetic disorder characterized by progressive aplastic anemia and an increased risk of developing various childhood cancers. The disease is caused by the loss of function in any one of at least 16 genes or complementation groups. We have developed a cell-based method for the identification of complementation groups in FA patients that utilizes retroviral-mediated gene transfer and subsequent correction of G2/M arrest in cells from the patient. Briefly, after transduction with the retroviral vectors, cells are exposed for 48 hours to the alkylating agent Melphalan and cell cycle analysis for FA-specific G2/M arrest is performed by flow cytometry. A complementation group is identified when correction in the G2/M arrest occurs. The TTDSL can assay for the following complementation groups: A, B, C, E, F, G, and L.
Due to the advent of the more widely used sequencing technologies, we no longer perform this test at a CAP level. However, we maintain the cell lines and reagents and ensure that we have trained personnel to perform the testing for research or investigational purposes. It is important to note that the technical procedure has not changed with the exception that the Sponsor must now select one complementation group to test. Additional complementation groups may be added at a reduced rate if requested at the same time. Contact us if you are interested in submitting a sample for this test.
1. Schambach A, Bohne J, Chandra S, Will E, Margison GP, Williams DA, Baum C (2006). Equal potency of gammaretroviral and lentiviral SIN vectors for expression of O6-methylguanine-DNA methyltransferase in hematopoietic cells. Mol Ther. 13(2):391-400
2. Will, E., Bailey, J, Schuesler, T., Modlich, U., Balcik, B., Burzynski, B., Witte, D., Layh-Schmitt, G., Rudolph, C., Schlegelberger, B., Von Kalle, C., Baum, C., Sorrentino, B.P., Wagner, L.M., Kelly, P., Reeves, L., and Williams, D.A. (2007). Importance of murine study design for testing toxicity of retroviral vectors in support of Phase I trials. Mol. Ther. ,Apr 15(4):782-91
3. Thornhill, S.I., Schambach, A., Howe, S.J., Ulaganathan, M., Grassman, E., Williams, D., Schiedlmeier, B., Sebire, N.J., Gaspar, H.B., Kinnon, C., Baum, C. and Thrasher, A.J. (2008). Self-inactivating gammaretroviral vectors for gene therapy of X-linked severe combined immunodeficiency. Mol Ther. 16(3):590-8
4. Zychlinski, D., Schambach, A., Modlich, U., Maetzig, T., Meyer, J., Grassman, E., Mishra, A. and Baum, C. (2008). Physiological promoters reduce the genotoxic risk of integrating gene vectors. Mol Ther. 16(4):718-25.
5. Nawrot M, McKenna DH, Sumstad D, McMannis JD, Szczepiorkowski ZM, Belfield H, Grassman E, Temples T, Nielsen D, Yuan N, Wognum B, Reems JA (2011). Interlaboratory assessment of a novel colony-forming unit assay: a multicenter study by the cellular team of Biomedical Excellence for Safer Transfusion (BEST) collaborative. Transfusion. 51(9):2001-5