Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy
Recent advances in recombinant adeno-associated virus (rAAV) gene therapy for choroideremia show gene replacement to be a promising approach. It is, however, well known that contact of vector solution with plastic materials in the surgical device may result in non-specific adsorption with resulting...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Cell Press
2019
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| _version_ | 1797073384636416000 |
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| author | Patrício, M Cox, C Blue, C Barnard, A Martinez-Fernandez De La Camara, C Maclaren, R |
| author_facet | Patrício, M Cox, C Blue, C Barnard, A Martinez-Fernandez De La Camara, C Maclaren, R |
| author_sort | Patrício, M |
| collection | OXFORD |
| description | Recent advances in recombinant adeno-associated virus (rAAV) gene therapy for choroideremia show gene replacement to be a promising approach. It is, however, well known that contact of vector solution with plastic materials in the surgical device may result in non-specific adsorption with resulting loss of physical titer and/or level of protein expression and activity. Here we assessed the biocompatibility and stability of rAAV2-REP1 (Rab Escort Protein-1) before and following passage through the injection device over a period of time to mimic the clinical scenario. Three identical devices were screened using two concentrations of vector: high (1E+12 DNase-resistant particles [DRP]/mL) and low (1E+11 DRP/mL), to mimic high- and low-dose administrations of vector product. The low dose was prepared using either formulation buffer that contained 0.001% of a non-ionic surfactant (PF68) or balanced salt solution (BSS). We observed significant losses in the genomic titer of samples diluted with BSS for all time points. The addition of 0.001% PF68 did not, however, affect rAAV physical titer, or REP1 protein expression and biological activity. Hence we observed that neither the genomic titer nor the biological activity of a rAAV2-REP1-containing solution was affected following passage through the surgical device when PF68 was present as a surfactant and this was maintained over a period up to 10 h. |
| first_indexed | 2024-03-06T23:21:20Z |
| format | Journal article |
| id | oxford-uuid:68d2ec3a-7659-49e5-8ece-743e4aa36dd9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T23:21:20Z |
| publishDate | 2019 |
| publisher | Cell Press |
| record_format | dspace |
| spelling | oxford-uuid:68d2ec3a-7659-49e5-8ece-743e4aa36dd92022-03-26T18:47:33ZInclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapyJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:68d2ec3a-7659-49e5-8ece-743e4aa36dd9EnglishSymplectic Elements at OxfordCell Press2019Patrício, MCox, CBlue, CBarnard, AMartinez-Fernandez De La Camara, CMaclaren, RRecent advances in recombinant adeno-associated virus (rAAV) gene therapy for choroideremia show gene replacement to be a promising approach. It is, however, well known that contact of vector solution with plastic materials in the surgical device may result in non-specific adsorption with resulting loss of physical titer and/or level of protein expression and activity. Here we assessed the biocompatibility and stability of rAAV2-REP1 (Rab Escort Protein-1) before and following passage through the injection device over a period of time to mimic the clinical scenario. Three identical devices were screened using two concentrations of vector: high (1E+12 DNase-resistant particles [DRP]/mL) and low (1E+11 DRP/mL), to mimic high- and low-dose administrations of vector product. The low dose was prepared using either formulation buffer that contained 0.001% of a non-ionic surfactant (PF68) or balanced salt solution (BSS). We observed significant losses in the genomic titer of samples diluted with BSS for all time points. The addition of 0.001% PF68 did not, however, affect rAAV physical titer, or REP1 protein expression and biological activity. Hence we observed that neither the genomic titer nor the biological activity of a rAAV2-REP1-containing solution was affected following passage through the surgical device when PF68 was present as a surfactant and this was maintained over a period up to 10 h. |
| spellingShingle | Patrício, M Cox, C Blue, C Barnard, A Martinez-Fernandez De La Camara, C Maclaren, R Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy |
| title | Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy |
| title_full | Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy |
| title_fullStr | Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy |
| title_full_unstemmed | Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy |
| title_short | Inclusion of PF68 surfactant improves stability of rAAV titer when passed through a surgical device used in retinal gene therapy |
| title_sort | inclusion of pf68 surfactant improves stability of raav titer when passed through a surgical device used in retinal gene therapy |
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