GENE THERAPY FOR LYSOSOMAL DISORDERS
Patent Information
- Application Number
- EA202490491
- Authority / Receiving Office
- EA · EA
- Patent Type
- Patents
- Current Assignee / Owner
- Priority Date
- 2020-03-12
- Filing Date
- 2020-04-10
- Publication Date
- 2026-09-23
- Estimated Expiration
- 2040-04-10
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Abstract
Claims
1. A method for the quantitative assessment of progranulin protein (PGRN) in a cerebrospinal fluid (CSF) sample from a subject known to have or suspected of having frontotemporal dementia (FTD), comprising: (a) dilution of the CSF sample in a master mix containing dithiothreitol (DTT) and sample buffer; (b) loading the diluted CSF sample, anti-PGRN antibody, a secondary antibody that detects anti-PGRN antibody, luminol, and peroxide into the wells of a capillary cartridge; (c) loading the capillary cartridge into an automated Western blot immunoassay instrument; (d) using an automated Western blot immunoassay instrument to calculate signal intensity, peak area, and signal-to-noise ratio; and (e) quantification of the PGRN protein level in the CSF sample as the peak area of immunoreactivity to the anti-PGRN antibody, where the CSF sample was taken from a subject who had previously been administered a recombinant adeno-associated virus (rAAV) containing: (i) an rAAV vector containing a nucleic acid comprising, in order from 5' to 3': (a) ITR of adeno-associated virus 2 (AAV2); (b) cytomegalovirus (CMV) enhancer; (c) chicken beta-actin promoter (CBA); (d) a transgene insert encoding a PGRN protein, wherein the transgene insert comprises the nucleotide sequence of SEQ ID NO: 68; (e) woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); (f) the polyA signal tail of bovine growth hormone; and (g) AAV2 ITR; and (ii) capsid protein of adeno-associated virus (AAV).
2. The method according to claim 1, wherein the CSF sample is taken from a human.
3. The method of claim 1, wherein the FTD is FTD with a GRN mutation.
4. The method of claim 1, wherein the CSF sample is taken from a non-human primate.
5. The method according to claim 1, wherein the AAV capsid protein is an AAV9 capsid protein.
6. The method according to any one of claims 1 to 5, wherein the dilution step is a 4-fold dilution.