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In AAV Research & Gene Therapy

Discover how Progen's range can help you with your experiments!

AAV

Immunochemical AAV studies

Anti-AAV virus particle antibodies are suitable for the characterisation of different stages of adeno-associated virus (AAV) infection and are very useful for the analysis of the AAV capsid assembly. The antibodies specifically recognize conformational epitopes in assembled capsids of different AAV serotypes. Hence, they exclusively react with intact AAV particles.

Viral capsid protein antibodies (VP) exclusively recognize AAV capsid proteins and are useful for immunolocalization studies of AAV capsid formation, or immunoprecipitation and western blot analysis of viral capsid proteins. Anti-AAV replicase antibodies (Rep) react with selected replicase (Rep) proteins in human AAV-infected cells. Applications include immunolocalization or immunoblotting studies to investigate the correlation between Rep expression and the course of an infection.

AAV antibodies in gene therapy research

AAV vectors are powerful tools in gene therapy research and development.  Recombinant AAV vectors (rAAV) corresponding to the different viral serotypes have successfully been used as universal gene shuttles in human cells.1 Neutralizing AAV antibodies present in serum or plasma may block transduction with AAV vectors.

In a prescreening step before vector administration, the titer of neutralizing AAV antibodies can be determined in a cell-based assay. PROGEN's AAV antibodies are ideal positive controls that neutralize wild-type AAV capsids of AAV serotypes 1, 2, 5, 6, 8, and 9.2-7

PROGEN supplements its portfolio of AAV antibodies with advanced recombinant IgGs that feature optimal stability and batch-to-batch consistency. These antibodies retain equal performance compared to the mouse monoclonal antibodies with respect to cross-reactivity, AAV capsid recognition and binding sensitivity (see figure below).

 

Comparison of PROGEN's A20 mouse monoclonal antibody with the recombinant A20R antibody (both against AAV2): A) Dot blot comparing cross-reactivities of A20 and A20R with different AAV serotypes. B) Dot blot analysis of antibody reactivity with denatured and native AAV2 capsids. C) Dot blot analysis of binding sensitivity of A20 and A20R by using decreasing AAV2 particle titers.

AAV Xpress ELISA

Now available for the serotypes AAV2, AAV8 and AAV9!

• Get robust & reliable results in less than 2 hours
• Enjoy easy workflow and process integration
• Rely on accurate data based on standard ELISA
• Simple processing – no expensive equipment needed
• Low sample costs
• Shortened incubation times from 3:15 hrs to 1:20 hrs
• Same composition of kit components
• Same processing
• Complete reduction of the assay time of more than 50% 
• Quantification of intact AAV capsids (full & empty)
• Accurate and reliable – low inter- and intra-assay variance
• Characterization of AAV preparations in combination with additional quantification methods, e.g. qPCR, ddPCR
• Widely used in industrial and academic labs using AAV vectors for the development of gene therapies

More information contact Info@2Bscientific.com

STANDARDIZED RESEARCH WITH AAV TITRATION ELISA

Reliable determination of AAV titers

The increasing interest in rAAV for clinical applications demands a dependable and reproducible quantification of accurate rAAV titers to ensure a safe and reliable gene transfer. In view of the scientific
and clinical significance, PROGEN has established a line of AAV quantification ELISAs for different AAV serotypes (1, 2, 3, 5, 6, 8, 9, and rh10), utilizing its portfolio of capsid-specific AAV antibodies.
The assays for the determination of rAAV2 and rAAV8 titers have been validated in international studies8, 1 and have been classified as superior method for a reliable, standardized rAAV particle titer quantification.

Serotype AAV1 AAV2 AAV2 AAV3
Cat. No. PRAAV1 PRATV PRAAV2R PRAAV3R
Serotype AAV5 AAV6 AAV8 AAV9 AAVrh10
Cat. No. PRAAV5 PRAAV6 PRAAV8 PRAAV9 PRAAV10

Development of internal AAV standards for ELISA kit calibration

The development of standardized AAV capsid material is an essential requirement for the calibration of all of PROGEN's AAV titration ELISA assays to ensure the reliable quantification of intact capsids in rAAV vector preparations. The alignment of the AAV2 and AAV8 Kit Controls for PROGEN's titration ELISAs is based on the ATCC standard material8, 9. For the remaining Kit Controls of the AAV1, 3, 5, 6 and 9 titration ELISAs PROGEN utilizes electron microscopy in combination with qPCR and ddPCR for the development of internal AAV gold standards (data for the development of the AAV5 standard shown below).


A) + B) Comparison of AAV5 sample and reference AAV8 (WL217S) capsids & distinction of full and empty capsids by EM: A) AAV5 micrograph.
B) AAV8 (WL217S) micrograph, which was used to characterize the current AAV8 RSM material from ATCC8. Samples were stained with uranyl acetate.
C) Comparison of AAV5 qPCR titers with ATCC standards for AAV2 and AAV8: A: 3 independent labs provided qPCR / ddPCR data for the AAV5 standard material. The mean titer of viral genome copies and CV was calculated. B: Published qPCR data for ATCC standard material AAV29. C: Published qPCR data for ATCC standard material AAV88.
D) Alignment of the Kit Control with the established gold standard in the AAV5 Titration ELISA: Aligned curves of gold standard material and Kit Control, measured with the AAV5 Titration ELISA (OD vs. concentration). Black: Curve of the gold standard material, Red: Curve of the Kit Control.