Recombinant AAV Genome Sequencing Service
Over the last decades, adeno-associated virus (AAV) has become one of the most actively investigated gene therapy vehicles. Particularly, recombinant AAV-based gene therapy has entered a phase of clinical translation and commercialization. As a leading company in the field of gene therapy, Creative Biolabs offers high-quality recombinant AAV genome sequencing service to help researchers assess vector purity and integrity for clinical application.
Importance of Recombinant AAV Genome Sequencing
Encouraging outcomes of clinical trials using vectors derived from recombinant AAV have helped to promote gene therapy for the treatment of genetic and acquired diseases. However, the production of recombinant AAV involves components of both cellular and viral origin, and despite extensive downstream purification, recombinant AAV production results in a heterogeneous product that is particularly complex to characterize. Therefore, rigorous quality control (QC) tests must be performed to ensure their efficiency and safety for human applications. Currently, standard QC tests are limited to the titration and quantification of a vector by qPCR analysis, verification of genome size by native or alkaline (denaturing) agarose-gel electrophoresis, and characterization of viral purity by silver-stained polyacrylamide gel electrophoresis. Unfortunately, these methods fall short since they cannot decipher the composition of individual vector genomes, making in-depth profiling of heterogeneous populations difficult.
Figure 1. Single-Particle Resolution Profiling of scAAV Genomes by AAV-Gpseq. (Tai, 2018)
In this condition, the recombinant AAV-gene therapy field is in need of new techniques that not only detect the encapsidation of undesirable genomes but can also offer clues to improve vector homogeneity. During the past, high-throughput sequencing platforms such as Helicos Biosciences single-molecule sequencing (SMS) and Illumina-based deep sequencing have been developed to profile packaged single-stranded (ss) AAV genomes to assess the extent of "error-prone" genome encapsidation during recombinant AAV production. Unfortunately, these methods also fall short of capturing fully intact vector sequences. Recently, AAV-genome population sequencing (AAV-GPseq) provides a new approach to profile vector heterogeneity with full-vector genome resolution.
Services
As a leading company in the gene therapy area, Creative Biolabs has successfully established high-throughput platforms for recombinant AAV genome sequencing. Particularly, we are capable of offering AAV-GPseq service to help global customers detect encapsidated, DNase I-resistant bacterial sequences originating from reverse packaging events, as well as detection of adenoviral helper and Rep/Cap-construct sequences packaged into virions. What's more, we also provide molecular characterization and quantitation of error-prone rAAV genome replication service for research-grade and clinical vector manufacturing QC pipelines.
Creative Biolabs is dedicated to providing novel genome sequencing services for recombinant AAV to help test their efficiency and safety. Please feel free to contact us for more details and we are pleased to offer you the best services.
Reference
- Tai, P.W.; et al. (2018). Adeno-associated virus genome population sequencing achieves full vector genome resolution and reveals human-vector chimeras. Molecular Therapy-Methods & Clinical Development. 9, pp.130-141.
