Status:
Advanced ReviewerMember since: 2020
Organization: Freeline Therapeutics
Great kit, it is part of our QC ELISA assays, highly recommended.
Application Area:Biotechnology, quality control
"It's quite a trustworthy ELISA kit, you get good performance in the assay and robust results. It's quite handy as all reagents, including controls, come with the kit."
Status:
ReviewerMember since: 2020
Organization: MilliporeSigma
Truly the best ELISA.
Application Area:Viral vectors
"I love the Progen titration ELISA. They are easy to use and give reliable, fast results. With the new Xpress ELISAs, they are even faster. Overall, I am a very happy customer."
Status:
ReviewerMember since: 2020
Organization: Pall Corp.
Excellent quality; our analytical method of choice to compare with other analytics.
Application Area:Analyze capsid titer in sample
"This kit is easy to use and the results are robust and reproducible. The quality control of the kit is excellent and yields results that are important to assessing the capsid quality in samples from bioreactors and other samples. Technical service is always knowledgeable and helpful with any questions or troubleshooting."
Status:
ReviewerMember since: 2020
Organization: King's College London
Consistent results and easy to use.
Application Area:Quantification of viral particles
"Ready to use ELISA kit for the titration of assembled AAV capsid particles that yields consistent and reliable titres across different samples and concentrations."
Status:
ReviewerMember since: 2018
Organization: Cobra Biologics
Fantastic product which is a core component of our R&D work into gene therapy.
Application Area:R&D into the production of rAAV for gene therapy. We are using this ~3 times a week and are very hap
"We use this product ~3 time per week and receive very reliable, consistent results; it is a critical part of our development within R&D that we cannot perform our work without. Alongside the AAV2 kit, we also use the AAV5 (PRAAV5) and AAV9 (PRAAV9) ELISA kits which we are also extremely satisfied with. Our after purchase care has also been exceptional with fast replies to any queries and opportunities to meet with the team."
The ELISA principle:
The assay is based on the sandwich ELISA technique where a monoclonal antibody (mab) specific for a conformational epitope on assembled AAV capsids is coated onto the plate and is used to capture AAV particles from the specimen.
The detection of captured AAV particles is a two-step process.
A biotin-conjugated mab is bound to the captured AAV particles.
A streptavidin peroxidase conjugate reacts with the biotin molecules. The addition of the substrate results in a color reaction which is proportional to the amount of specifically bound viral particles.
Comparison of AAV Quantification Methods:
Each of the commonly used quantification methods has its pros and cons:
Given the practical drawbacks of the aforementioned techniques, a conventional sandwich ELISA currently appears to be superior in terms of inter- and intralaboratory variation as well as ease of use. Therefore, it represents the best format for reliable and reproducible quantification of total rAVV capsid titers.
Use of shuffled/mutated AAV:
The recognition of shuffled/mutated AAV vectors depend on the specific capsid region which is affected by the shuffling/mutation. The capture antibodies used for PROGEN's AAV ELISAs bind specific and, in some cases, well defined conformational epitopes. These epitopes are generated by the capsid assembly of the corresponding AAV serotypes. A first indication that the ELISA might recognize your shuffled/mutated AAV vector is the presence of the antibody-binding epitope. However, changes in the protein sequences of the capsid proteins might also influence conformation of the proteins, hence the conformation of the epitopes presented on the AAV capsid. This might influence binding affinity of the antibody and affect determination of the titer based on the (non-shuffled) Kit Control provided with the AAV ELISA kit. Since these properties strongly depend on the specific shuffling/mutation performed, PROGEN cannot guarantee successful and precise quantification of your shuffled/mutated AAV vector. Even if the antibody binding epitopes are still present on your shuffled/mutated AAV capsid, your assay needs to be tested and optimized for your specific AAV vector.PR
OGEN highly recommends the production and calibration of a suitable (shuffled/mutated) Kit Control, to ensure reliable titer determination of your individually shuffled/mutated AAV vector with PROGEN ELISA kits.
Limited Use Label License: Research Use OnlyProduct is exclusively licensed to PROGEN Biotechnik GmbH. The use of these products for the development, manufacturing and sale of secondary products/derivativeswhich are based on the purchased products and/or which include the purchased product require a royalty based sub-license agreement.