Show simple item record

dc.contributor.authorBennett, Antonette
dc.contributor.authorPatel, Saajan
dc.contributor.authorMietzsch, Mario
dc.contributor.authorJose, Ariana
dc.contributor.authorLins-Austin, Bridget
dc.contributor.authorYu, Jennifer C.
dc.contributor.authorBothner, Brian
dc.contributor.authorMcKenna, Robert
dc.contributor.authorAgbandje-McKenna, Mavis
dc.date.accessioned2017-10-16T14:23:25Z
dc.date.available2017-10-16T14:23:25Z
dc.date.issued2017-09
dc.identifier.citationBennett, Antonette, Saajan Patel, Mario Mietzsch, Ariana Jose, Bridget Lins-Austin, Jennifer C. Yu, Brian Bothner, Robert McKenna, and Mavis Agbandje-McKenna. "Thermal stability as a determinant of AAV serotype identity." Molecular Therapy - Methods & Clinical Development 6, no. 15 (September 2017): 171-182. DOI:https://dx.doi.org/10.1016/j.omtm.2017.07.003.en_US
dc.identifier.issn2329-0501
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/13813
dc.description.abstractCurrently, there are over 150 ongoing clinical trials utilizing adeno-associated viruses (AAVs) to target various genetic diseases, including hemophilia (AAV2 and AAV8), congenital heart failure (AAV1 and AAV6), cystic fibrosis (AAV2), rheumatoid arthritis (AAV2), and Batten disease (AAVrh.10). Prior to patient administration, AAV vectors must have their serotype, concentration, purity, and stability confirmed. Here, we report the application of differential scanning fluorimetry (DSF) as a good manufacturing practice (GMP) capable of determining the melting temperature (Tm) for AAV serotype identification. This is a simple, rapid, cost effective, and robust method utilizing small amounts of purified AAV capsids (∼25 μL of ∼1011 particles). AAV1-9 and AAVrh.10 exhibit specific Tms in buffer formulations commonly used in clinical trials. Notably, AAV2 and AAV3, which are the least stable, have varied Tms, whereas AAV5, the most stable, has a narrow Tm range in the different buffers, respectively. Vector stability was dictated by VP3 only, specifically, the ratio of basic/acidic amino acids, and was independent of VP1 and VP2 content or the genome packaged. Furthermore, stability of recombinant AAVs differing by a single basic or acidic amino acid residue are distinguishable. Hence, AAV DSF profiles can serve as a robust method for serotype identification of clinical vectors.en_US
dc.description.sponsorshipNational Institutes of Healthen_US
dc.titleThermal stability as a determinant of AAV serotype identityen_US
mus.citation.extentfirstpage171en_US
mus.citation.extentlastpage182en_US
mus.citation.issue15en_US
mus.citation.journaltitleMolecular Therapy - Methods & Clinical Developmenten_US
mus.citation.volume6en_US
mus.identifier.categoryLife Sciences & Earth Sciencesen_US
mus.identifier.doi10.1016/j.omtm.2017.07.003en_US
mus.relation.collegeCollege of Letters & Scienceen_US
mus.relation.departmentChemistry & Biochemistry.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.data.thumbpage3en_US


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record