Immunoassays are an essential bioanalysis tool used throughout cell and gene therapy development and manufacturing to evaluate viral vector titer and immunogenicity responses to gene therapy treatment. Titer analysis for the two most commonly used vehicles for introducing genetic therapies into mammalian cells, adeno-associated virus (AAV) and lentivirus vectors is commonly through time-consuming ELISA assays for AAV titer and lentiviral titer.
Utilization of robust and innovative immunoassay platforms for bioanalysis in gene therapy development can streamline production. Gyrolab® automated microfluidic, nanoliter-scale immunoassay solutions dramatically shorten time to results to one hour, reduce sample volumes, and improve data quality for AAV- and lentivirus-based gene therapy immunoassays during development, and are ideally suited for compressed cell and gene therapy development timelines.
BIOPROCESSING GYROLAB ASSAYS
PRECLINICAL AND CLINICAL STUDY GYROLAB ASSAYS
Oxford Biomedica has established itself on the gene therapy stage, being the first to administer lentiviral vectors in vivo and as the first lentiviral vector containing product to gain approval for advanced therapy in the US using a unique LentiVector® enabled technology (Novartis Kymriah®). Over 500 patients have been treated with gene-based therapeutics by Oxford Biomedica or its partners, and a number of new therapies are in development. The team at Oxford Biomedica saw that by using the Gyrolab system they would be able to run an assay 24X faster and with 10X less sample compared to using ELISA. See blog post, "Putting a New Spin on Immunoassays for Gene Therapy - Part 1"
Cell and gene therapy development demands for speed, precision, reproducibility, and GxP compliance in bioanalytical methods are met with Gyrolab microfluidic, CD-based assay technology.
Centrifugal force and a flow-through affinity microcolumn format eliminate lengthy incubations and minimize matrix interference. The nanoliter-scale format saves reagents and samples, resulting in a cost-effective and highly reproducible immunoassay that produces quality data over broad dynamic ranges. Gyrolab BioaffyTM CDs combined with Gyrolab automated systems dramatically reduce hands-on time and shorten processing times, leading to considerable gains in productivity.
Advancing new cell and gene therapies through preclinical and clinical studies requires specialized expertise in regulatory requirements, study design and execution, and scale-up vector manufacturing technologies. Many pharma and biotech companies secure outsourcing CRO and CMO partners for this purpose. CRO/CMO partners face additional timeline and quality demands for immunoassay workflows when conducting GLP- and GMP-compliant studies, often with fixed timelines specified by commercial sponsors. Gyrolab immunoassays meet stringent criteria for regulatory-compliant CRO or CMO bioanalysis:
The measurement of anti-vector or transgene antibodies using a robust and reproducible immunoassay is an integral bioanalytical component in preclinical gene therapy studies to understand the potential for immunogenic reactions in human clinical use. Measurement of pre-existing anti-drug antibodies (ADA) may also be used to pre-screen subjects in clinical studies to avoid neutralizing antibodies potentially affecting efficacy. Cytokine biomarker immunoassays may also be used as an indicator of an inflammatory innate immune response to indicate the potential for a “cytokine storm” response. Gyrolab ADA software guides the development of automated immunoassays for ADA analysis.
Gyrolab preclinical and clinical study immunoassays:
Quantifying functional viral titer for lentiviral or AAV vector production is time-consuming and costly. Getting results fast is important to respond quickly and adjust culture or bioprocess conditions to optimize vector titer.
Minimizing co-purifying impurities of host cell proteins (HCPs) that are potentially toxic or immunogenic is important during manufacturing. HCP measurement is a critical regulatory requirement and having an open platform that has the flexibility to measure HCPs from new expression host systems or utilizing in-house developed assays is essential. Gyrolab kits and assay protocols are available for a wide range of gene therapy bioprocess analytics:
Viral vector titer and characterization
Host Cell Protein (HCP) impurity analysis
Cell culture and other process-related impurities
Gyrolab p24 Kits, Gyrolab AAVX Kits, and Gyrolab AAV9 Kits provide plug and play, miniaturized lentiviral p24 titer, and AAV titer immunoassays for use on all Gyrolab platforms. Kits include everything needed to generate 96 or 4800 data points, including capture reagent, detection reagent, standard curve samples, wash buffer, sample dilution buffer, and Gyrolab Bioaffy CD(s). Used together with Gyrolab systems, 96 data points are delivered in 80 minutes to accelerate lentiviral vector bioanalysis during production and bioprocessing.
Gyrolab systems meet all these criteria for advanced gene therapy bioanalytical assay performance and speed needed to succeed in cell and gene therapy development. Gyrolab systems are well established in pharma and both preclinical and clinical CROs and support a wide range of applications including immunogenicity, IgG titer, HCP analysis, and viral vector titer.