Simple Plex™ Assays for Viral Titer Quantification
Fast and reproducible viral capsid measurement is essential for those focused on gene transfer, as part of a cell and gene therapy workflow. While traditional plate-based and dot blot assays can be useful, they are often complex, time-consuming, and non-standardized. See how Simple Plex assays on the EllaTM platform are used for viral titer quantification. Next, meet Ella and request pricing. Finally, choose the Simple Plex Assay that best meets your needs.
Table of Contents
- Webinar: Scaling Analytics for Cell and Gene Therapy With Ella
- Comparing the Simple Plex AAV Assays With the PROGEN AAV ELISAs
- Select A Simple Plex AAV Assay: AAV1, AAV2, AAV6, and AAV8
- Lentiviral Vector Titration
- Find Out About Bio-Techne's Tools For Cell And Gene Therapy
- View Complete Simple Plex Assay Menu
Viral titer with Simple Plex Immunoassays
Understanding the number of particles, or infectious units, in your viral prep is essential, as it directly determines the viral transduction efficiency (i.e., how effectively the viruses will deliver genetic material into the target cells) and the resulting number of integrated gene copies. Accurate viral titration is a cornerstone of viral transduction workflows, ensuring that sufficient viral preps have been performed to achieve the desired transduction efficiency and generate reliable results during cell transfection experiments.
Simple Plex Viral Titer assays, available on the Ella platform, provide a fast, reliable, and reproducible solution for viral titration. These assays streamline the process, minimizing variability while delivering precise quantification of viral capsid proteins across a wide dynamic range. Whether you are working with adeno-associated viruses (AAV) or lentivirus, Simple Plex offers assays tailored for both, making it easier to optimize workflows and ensure the success of your gene delivery or cell therapy applications.
Simple Plex AAV Assays are paired with PROGEN AAV Antibodies
For application of AAV vectors in gene therapy, the determination of total capsid titer is essential. PROGEN AAV Titration ELISAs are well-known for their ability to provide reliable and accurate quantification of AAV total capsid titers, based on the proven sensitivity and specificity of PROGEN AAV antibodies for AAV intact particles. The Simple Plex AAV assays offer the same proven specificity as the industry-leading PROGEN ELISA along with the convenience, sample-conserving, and timesaving advantages inherent in the fully automated Ella platform.
All steps of the Simple Plex AAV assays on Ella are highly automated thanks to the microfluidic cartridge—everything is preloaded, even the calibration curve. Just pipette your diluted samples onto the cartridge, add wash buffer, press start in the Ella Runner Software. In addition, each Simple Plex AAV cartridge includes a fully built-in and pre-validated standard curve. This eliminates the need to prepare, plate, or serially dilute standards—reducing hands-on time, removing a major source of operator variability, and helping ensure consistent, reproducible quantitation across assays, cartridges, and users.
Inside the microfluidic cartridge are glass nano reactors (GNRs), the core of the Simple Plex Assay. In Simple Plex AAV Assays, the GNRs are coated with the PROGEN antibodies that work to capture intact AAV capsids. That same corresponding AAV antibody is also lyophilized in the cartridge to act as the detection reagent. The Ella Platform’s fluorescence detection system enables detection across a broad dynamic range, minimizing the need to run multiple dilutions to find the ideal MRD. The efficient cartridge design, featuring three independent GNRs per sample inlet, eliminates the need to run samples in duplicate. In total, from set up to fully processed and analyzed results takes less than 90 minutes.
Creating Lentiviral Vectors With Fast, Reliable Viral Titration
With the growing use of lentiviral vectors derived from HIV-1 in gene transfer workflows, researchers need fast, reliable viral titer quantification during process development. The 3-to-6-hour timeframe required to run a standard ELISA is a challenge for operators who increasingly require results quickly to inform real-time decision-making.
The Simple Plex assay for the detection of human HIV-1 Gag p24 combines high throughput with superior accuracy and precision to enable rapid quantification of lentiviral capsid proteins throughout different stages of cell and vector manufacturing. The automated, reproducible nature of the Simple Plex p24 assay helps eliminate user error, making it an ideal tool in good manufacturing practice (GMP) environments where quality, standardization of process, assay transfer, and scalability are highly desirable.
Linear correlations of Simple Plex AAV assays and PROGEN AAV ELISAs. The aggregated Simple Plex and PROGEN ELISA data was plotted and a linear fit was applied to the data for each AAV serotype. The Simple Plex AAV assays are corelated with the corresponding PROGEN ELISAs with an R2=0.99 (2a-2d). The data shows the quantitative range of both assays overlap.
Bio-Techne's Tools For Cell And Gene Therapy
Bio-Techne offers a diverse portfolio of products and services designed to accelerate and simplify your cell and gene therapy workflow. From GMP proteins and small molecules to state-of-the-art analytical instrumentation, Bio-Techne is committed to providing high quality innovative solutions. Learn more about Bio-Techne’s cell and gene therapy offerings.
In a multi-use study conducted by 7 operators on 11 Ella instruments, the Simple Plex HIV-1 p24 assay demonstrated great precision and reproducibility. The automated, microfluidic design of the Simple Plex assays running on Ella helps eliminate user-introduced variability by offering built-in standard curves and separate channels for each analyte, which also ensures no cross-reactivity. The results for the low control and high control intra and inter assays were reproducible with an overall coefficient of variation (CV) between 2% and 11%.
The Simple Plex HIV-1 p24 assay has demonstrated impressive linearity to showcase the accuracy within the expected assay range. The results for the five diluted points show that diluted samples are able to provide reliable results (Figure 3). The automated cartridge is designed to eliminate the lengthy incubations typical of traditional plate-based immunoassays while ensuring precise and consistent results across multiple users.
Ella satisfies the demands of timely data generation for maintaining efficient production and overcomes the lengthy investigations and sample retests that often result from human error. You also get excellent assay sensitivity and highly reproducible results, which helps ensure you get the high-quality data required to meet regulatory standards for titration throughout the viral vector production workflow.
Documents
Application Note: Simple Plex AAV Assays
Application Note: Simple Plex AAV Assays
In this application note, we demonstrate assay comparability between the Simple Plex AAV assays and the PROGEN AAV Titration ELISA Kits.
Brochure: Meet Ella
Brochure: Meet Ella
Ella is your immunoassay problem solver. Simple Plex assays are hands-free, single or multi-analyte with no cross-reactivity, and reproducible even across multiple labs.
eBook: Analytical Technologies for Gene Therapy
eBook: Analytical Technologies for Gene Therapy
Inside this eBook, explore how our analytical instruments support multiple critical quality attributes from development to clinic.
What are viral titer assays?
Viral titer assays are designed to quantify viral vector concentration. In addition to cell and gene therapy gene transfer workflows, viral titer assays are integral to the study of infectious disease pathogenesis and vaccine development. At a high level, viral titer assays quantify: DNA titer, infectious titer, or capsid titer. DNA titer assays quantify the number of viral capsids containing DNA. Infectious titer assays quantify infectious particles that transfer DNA into host cells via a process called viral transduction. Capsid titer assays quantify viral capsids. Capsids may or may not contain viral DNA.
Learn more about viral titer assays at our Partner PROGEN’s website
What type of viral titer assay is the Simple Plex Immunoassay?
Simple Plex automated viral titer assays quantify viral particles, also known as capsids.
Learn more about Simple Plex Assays and the Ella Platform
I am looking for automated and scalable solutions for cell and gene therapy research and manufacturing. What do you suggest?
ProteinSimple, a Bio-techne Brand has several analytical solutions to suit your cell and gene therapy research and development needs. Download our eBook to see real-world examples of time, cost, and sample savings from our analytical solutions.
Download the Next Generation Analytical Solutions for Cell and Gene Therapy eBook
Do you have any other resources for cell and gene therapy research and manufacturing?
Bio-Techne spans the cell and gene therapy workflow. Solutions are available for immune cell therapy and regenerative medicine. Gene engineering services are also available using our nonviral TC Buster™ gene delivery system. GMP-grade raw materials and services are also available.
Find out more about our solutions for cell and gene therapy research and manufacturing at Bio-Techne.