Genome Plasmid

Description

We are planning a clinical trial to treat a rare genetic disease that causes progressive dementia in children. Our therapy is based on the following: Autologous haematopoietic stem progenitor cells (HSPCs) are modified ex vivo with a lentiviral vector (LV) to express supraphysiologic levels of a secreted enzyme in myeloid progeny, in essence using gene-modified monocytes/macrophages as a durable and effective vehicle for central nervous system (CNS) enzyme delivery. In order to progress this therapy, we require the following; a plasmid to our specification, produced to FDA, IND ready standard for use in the manufacture of a lentiviral vector for clinical trial. This Plasmid will be delivered directly from the supplier to the University of Edinburgh’s partner in the project, University of California Los Angeles. Lot 1: We are planning a clinical trial to treat a rare genetic disease that causes progressive dementia in children. Our therapy is based on the following: Autologous haematopoietic stem progenitor cells (HSPCs) are modified ex vivo with a lentiviral vector (LV) to express supraphysiologic levels of a secreted enzyme in myeloid progeny, in essence using gene-modified monocytes/macrophages as a durable and effective vehicle for central nervous system (CNS) enzyme delivery. In order to progress this therapy, we require the following; a plasmid to our specification, produced to FDA, IND ready standard for use in the manufacture of a lentiviral vector for clinical trial. This Plasmid will be delivered directly from the supplier to the University of Edinburgh’s partner in the project, University of California Los Angeles. This will require the following QC to be performed: - Appearance - Clear, Colorless, No Visible Particulates upon visual inspection - A260:280 - 1.7-2.0 measured by Spectrophotometer - Concentration - 1.0 - 1.1 mg/ml (0.95-1.14mg/ml) measured by Spectrophotometer - Endotoxin - <30 EU/mg measured by Endosafe® PTS - DNA Homogeneity - ≥90% All Supercoiled Forms analysed by Agarose Gel Electrophoresis - Residual RNA - None Detected @ 1.0mg load analysed by Agarose Gel Electrophoresis and Plasmid Safe™ Assay - Residual ssDNA - None Detected @ 1.0mg load analysed by Agarose Gel Electrophoresis and Plasmid Safe™ Assay - Residual Chromosomal DNA - None Detected @ 1.0mg load analysed by Agarose Gel Electrophoresis and Plasmid Safe™ Assay - Residual Linear DNA - <3% analysed by Plasmid Safe™ Assay and Linear DNA Assay - Identity - Consistent with Provided Information for 4 Restriction Reactions with no Discrepancies analysed by Restriction Digest and Research Primer Walking Sequencing - Protein – Reported measured by Micro BCA™ - pH of Final Product - 7.5-8.5 measured by pH meter - Bioburden - No growth on plates analysed by Five Day Bioburden The supplier must have previously supplied a plasmid to this specification that has been used in a similar clinical trial. The supplier must be able to evidence prior success in FDA approvals where their product has been used. The supplier will be required to deliver the product no later than 2 months after contract signature Supplied should follow all relevant legislation applicable to location of operation and State of California, USA