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Chapters
- 00:05Title
- 00:35Site-directed Mutagenesis
- 01:45Transformation and Screening for the Desired Clone
- 02:35pcDNA3.1/GLA and pcDNA3.1/GAA Plasmid Transfection and Treatment of HEK293H
- 04:00Cell Harvest
- 04:43Homogenization by Freezing and Thawing and BCA Assay
- 06:17Enzyme Activity Measurement with Artificial 4-Methylumbeliferyl Substrate
- 08:19Results: Enzyme Activity Measurement Mutagenized 3 α-galactosidase and 3 Acid α-glucosidase
- 09:35Conclusion
Det finns en efterfrågan att göra prekliniska test för en ny klass av ”särläkemedel” kallas farmakologiska chaperones reproducerbara, snabb och effektiv. Vi utvecklat en enkel, mycket standardiserad och mångsidig cell kultur-baserad analys till skärmen för kvalificerade patienter samt nya farmakologiska förkläde droger.
Tags
In VitroEnzyme MeasurementPharmacological ChaperoneFabry DiseasePompe DiseasePhenotypic AssessmentAllelic VarianceLysosomal Storage DiseasesPrognostic OutcomesTherapeutic DecisionsSite-directed MutagenesisGLA GeneGAA GenePrimer DesignPCRDpn1E.coliPlasmid PurificationSpectrophotometerHEK293H CellsCell Transfection
