Intrinsic differential scanning fluorimetry for fast and easy identification of Adeno-Associated Virus serotypes

Rieser R, Penaud-Budloo M, Bouzelha M, et al.

Journal of Pharmaceutical Sciences 2020, vol: 109(1) doi: 10.1016/j.xphs.2019.10.031

Abstract

Recombinant adeno-associated virus (AAV) vectors have evolved as the most promising technology for gene therapy due to their good safety profile, high transduction efficacy, and long-term gene expression in non-dividing cells. AAV-based gene therapy holds great promise for treating genetic disorders like inherited blindness, muscular atrophy, or bleeding disorders. Multiple naturally occurring and engineered AAV serotypes exist, which differ in capsid sequence and as a consequence in cellular tropism. Individual AAV capsids differ in thermal stability and have a characteristic melting temperature (Tm), which enables serotype-specific discrimination of AAV vectors. Differential scanning fluorimetry (DSF) combined with a dye-like SYPRO Orange (SO-DSF), which binds to hydrophobic regions of unfolded proteins, has been successfully applied to determine the Tm of AAV capsids. Here, we present DSF measurement of intrinsic fluorescence signal (iDSF) as a simple alternative method for determination of AAV capsid Tm. The study demonstrates that DSF measurement of intrinsic fluorescence signal is a simple, accurate, and rapid alternative to SO-DSF, which enables characterization of AAV capsid stability with excellent precision and without the need of SO or any other dye.

View Publication
 

Topics: Gene Therapy, Prometheus – nanoDSF,  Publications

 

 

 

Previous Article
Antibody-targeted chromatin enables effective intracellular delivery and functionality of CRISPR/Cas9 expression plasmids
Antibody-targeted chromatin enables effective intracellular delivery and functionality of CRISPR/Cas9 expression plasmids

Up next
Humanized Selective CD19CAR-T Cells Treatment Shows Efficacy in B-ALL Patients Who Relapsed after Receiving Murine-Based CD19CAR-T Therapies
Humanized Selective CD19CAR-T Cells Treatment Shows Efficacy in B-ALL Patients Who Relapsed after Receiving Murine-Based CD19CAR-T Therapies

Want to see more
gene therapy content?

Explore resources