Vaccine Development
Vaccines represent the best hope for COVID-19 management. The pandemic’s unprecedented health and economic impact are driving innovation to rapidly develop, evaluate, and produce a vaccine on a global scale. This challenge has led to a remarkable number of candidate vaccine platforms including, next-generation vaccine technologies. Nucleic acid (DNA or RNA) and viral vectors may offer unique scalability and efficacy, while proven approaches using recombinant protein or live attenuated/inactivated viruses are also in development. Regardless of the vaccine platform, expediency and scale-up are key, and choosing the best tools and technology are crucial. Bio-Rad offers solutions to advance your development and expedite scale-up for your fight against COVID-19.
Cell Line Development
Gene-edit Screening
Stable high-producing cell lines are fundamental to produce vaccines at a scale to fulfill global demand. Cell line engineering holds immense potential for rapid cell line development. Targeted technologies like CRISPR gene editing, zinc-finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN) offer tremendous potential in cell line development. Due to the low efficiency of gene editing, accompanying sensitive screening is necessary.
Droplet Digital PCR (ddPCR) complements cell line engineering by providing sensitive detection of positive clones as well as characterizing the expected frequency of homologous recombination. When paired with gene-editing technology, ddPCR enables sensitive streamlined approaches in cell line development.
Genetic Stability
The genetic stability of cell lines demonstrates quality and safety over time. Limitations in precision for gene copy number assessment can cause uncertainty during characterization and expansion, which can lead to wasted time and resources.
Droplet Digital PCR offers the ability to confidently and reproducibly determine the gene vector copy number to assess the stability and safety of your cell lines to assure productivity.
Cell Screening for Clonal Selection
Creating highly-productive cell lines involves significant screening, with one important parameter being cell-specific productivity. Analysis methods that enable the characterization of an individual cell’s phenotype can identify characteristics and culture conditions that allow for optimal cell-specific productivity. It is crucial to identify clones for production to expedite vaccine candidate selection readily.
The ZE5 Cell Analyzer provides speed, sensitivity, sample handling flexibility, and high-parameter multiplexing, all while offering maximal productivity.
Increase Speed
Analyze 384 well plate in less than 60 min with acquisition speed of 100K events/sec
Gain Versatility
Integrated sample loader to easily switch between tubes and plates
Maximize Throughout
Application programing interface (API) available for automation integration and 24/7 operation
Learn how to discover more with flow cytometry, click here
ZE5: A Better Solution for Automated High-Speed and High-Throughput Needs
This webinar is focused on the ZE5 Cell Analyzer's innovative designs in hardware and software to empower its automation and high-throughput capabilities. Its smart technology and automation-readiness make the ZE5 an ideal solution for automated high-throughput flow cytometry needs.
Immune-mediated Response
Cell Phenotyping
A significant challenge in vaccine development is characterizing the cellular immune response and identifying the key mediator of efficacy. These cell phenotypes can be complex, but essential for researchers to glean insights into what they may observe in preclinical and clinical studies. These multiplex, large-scale assays can benefit from automated high-throughput, high-parameter flow cytometry to identify treatments faster.
The ZE5 allows for high-throughput, high-parameter immunophenotyping to identify and characterize immune responses as a result of vaccine treatment.
Physical Viral Vaccine Titer
Exact viral titration of vector genome copies is critical for ensuring accurate and reproducible vaccine dosing in both preclinical and clinical settings. ddPCR directly quantifies DNA copies with exemplary precision without a standard curve, thereby reducing the challenges associated with choosing an appropriate, consistent standard. The absolute nature of the measurement results in a significantly lower coefficient of variation (<10% CV) compared to qPCR across both in-process and purified samples, allowing for consistency across all manufacturing stages.
Bio-Rad’s ddPCR systems utilize a simple workflow with minimal hands-on time while reducing sample replicates and user-to-user variability. Droplet Digital PCR is GMP-ready, designed to meet production and QC environment needs:
- Regulatory edition software to assist with 21 CFR Part 11 compliance
- Installation qualification/operational qualification (IQ/OQ) service
- Premier service offerings with rapid on-site support
Potency
Among the critical quality attributes, potency ensures vaccine efficacy. Potency is determined based on individual product attributes. The unrivaled precision of ddPCR offers the ability to sensitively measure integrated viral vector vaccine integration or DNA vaccine transfection efficiency. Complementary to cell integration confirmation, mRNA transgene expression can likewise be quantified, regardless of the sample matrix due to the inhibitor tolerance of ddPCR.
Impurities Detection
The FDA requires the reporting and quantification of host cell protein (HCP) and host cell DNA (HCD) in biologics since it can affect the efficacy and toxicity of the final product. Levels of these materials are critically monitored during process development and validation.
The GS-900 Calibrated Densitometer is validated and designed to quantitate the levels of HCP impurities.
Biologics Analysis WorkflowFlexible
Compatible with a wide variety of protein stains including Coomassie and silver stain
Accurate
Internal calibration using NIST-traceable step tablets
GMP Compliant
Automated analysis using Image Lab Software with 21 CFR Part 11 compliance with available installation qualification/operational qualification (IQ/OQ) protocols
Droplet Digital PCR provides a highly sensitive method for direct quantification of residual HCD to ensure adherence to the FDA guideline of < 100 pg/dose.
- No DNA purification required
- Inhibitor tolerance to in-process low pH and high salt sample matrices
- Available optimized supermix free of detectable E. coli, CHO, mouse, human and yeast DNA
Supporting publications:
Method for Residual DNA Quantification that Eliminates DNA Extraction, Protease Digestion and Use of DNA Standards
Biodistribution
Biodistribution analysis is a fundamental component of pharmacokinetics and must be carried out prior to clinical trial application in accordance with the FDA. Ideally, vaccines should persist long enough to stimulate antibody production but be cleared without long-term persistence. This is particularly necessary for next-generation vaccines that have limited track records, such as nucleic acid vaccines. For these nucleic acid vaccines, long-term persistence of vector DNA bears tumorigenic potential, while RNA vaccines typically suffer from instability. A thorough biodistribution evaluation is necessary to understand the safety and efficacy profile of a vaccine.
Bio-Rad offers high-performance CFX Real-Time systems and reagents for qPCR, the gold standard in biodistribution analysis. For added sensitivity and reproducibility to meet FDA biodistribution assay requirements, Bio-Rad offers ddPCR systems and reagents.
Supporting publications:
CFX qPCR Absolute Quantification of Plasmid DNA in Biodistribution StudiesViral Load Monitoring
Viral load is the measurement of viral particles in a patient. During clinical trials, monitoring the viral load is essential in indicating the efficacy of a vaccine against SARS-CoV-2. The precise quantification of viral particles is a critical indicator of patient response.
Droplet Digital PCR provides an ultra-sensitive and highly-precise absolute quantification of nucleic acid sequences. The sensitivity and precision ensure reproducible results with minimal lab-to-lab variability. ddPCR measures absolute quantities by counting nucleic acid molecules encapsulated in discrete and volumetrically defined water-in-oil droplet partitions. Droplet Digital PCR is particularly useful for low-abundance targets, targets in complex backgrounds, and monitoring subtle changes in target levels undetectable by real-time PCR.
The SARS-CoV-2 ddPCR test is based on highly sensitive and precise Droplet Digital PCR technology and is optimized for use on Bio-Rad Droplet Digital PCR Systems.
- High sensitivity and precision in low viral abundance samples
- Resistant to inhibition often seen in RT-PCR testing
- Single-well test — 3 sequences aligned to CDC markers: SARS-CoV-2 N1 and N2 genes, human RPP30 gene
Supporting publications:
ddPCR Improves Sensitivity and Robustness of Viral Load MeasurementsUsing Droplet Digital PCR to Quantify Levels of HIV Reservoirs
Dr Gary Lee, Senior Scientist at Sangamo BioSciences, Inc., and his team use Bio-Rad's Droplet Digital PCR system to quantify the level of HIV reservoirs in patients treated with their unique zinc-finger-based HIV therapy. With the need to detect approximately one copy of HIV in 1,000 cells analyzed, this quantification would present a challenge for conventional methods.