Understanding AAV¹

The adeno-associated virus (AAV) is a small (25 nm), non-enveloped virus of the parvoviridae family, including 12 different AAV serotypes. In the parvoviridae family it belongs to the genus dependoparvovirus, because it needs the presence of a helper virus for replication and assembly. The icosahedral AAV capsid composed of the capsid proteins VP1, VP2 and VP3 contains a linear, single-stranded DNA genome of 4.7 kb.

The AAV genome is flanked by two inverted terminal repeats (ITRs) and consists of two open reading frames (ORF): the rep and cap ORF. The ITRs are 145bp palindromes, GC rich and essential for packaging the viral DNA, replication, transcription and site-specific integration. The rep gene encodes the four rep proteins: rep78, rep68, rep52, rep40, which are expressed with the help of two promoters and alternative splicing. The rep proteins are essential for DNA replication, packaging the viral genome and its integration into the host DNA.

The viral capsid proteins VP1, VP2 and VP3 (87kDa, 72kDa, 62kDa) are encoded by the cap gene containing three alternative start codons, one for each of the capsid proteins. The AAV capsid is formed by a composition of 60 VP proteins in a 1:1:10 ratio (VP1:VP2:VP3). Cap also codes for the assembly activating protein AAP and membrane-associated accessory protein MAAP.

¹Source: Progen 

AAV ELISAs

Enzyme Immunoassay for the Quantitative Determination of AAV Serotypes 1, 2, 3, 5, 6, 8, 9, and rh10 Particles in Cell Culture Supernatants and Purified Virus Preparations. Immunotitration by PROGEN's AAV Xpress ELISA & AAV Titration ELISA offers a fast, sensitive and reproducible method for titration of intact AAV wild type virions, AAV recombinant virions or assembled and intact empty AAV capsids.

AAV Antibodies of Progen

PROGEN antibodies are useful for the characterization of different stages of an AAV infection and effective in the analyses of the AAV assembly process. There are two main categories of AAV antibodies:

  1. AAV capsid protein antibodies that specifically react with denatured AAV capsid proteins and thereby recognize the linear epitopes of the viral capsids.
  2. AAV particle antibodies that specifically react with intact AAV particles, i.e. empty and full capsids and thereby recognize a conformational epitope of the assembled capsids of certain serotypes.

PROGEN also offers a selection of AAV replicase antibodies.

Using AAV Antibodies for AAV-based Gene Therapy

This short video gives an overview of capsid and particle antibodies and how they can be used in your next AAV gene therapy project. 

Custom AAV packaging service

Sequence and expression-validated cDNA clones (TrueORF), ready for AAV packaging. A service of Origene Technologies.

AAV Purification Kit

Norgen’s AAV Purification Kit provides a fast and simple procedure for concentrating and purifying AAV vectors from cell lysate and cell culture media. Specifications:

  • AAV Purification from any input – cell fraction or media fraction
  • Purification from 500 mL of cell culture supernatant or from ~15 mL of cell pellet per kit
  • Rapid purification within 2 to 4.5 hours
  • High AAV recovery, up to 90%
  • No specialized equipment needed
  • Purification from a variety of AAV serotypes (including AAV6 and AAV9)
  • Purify AAV cell culture supernatant from 1 mL to 33.5 mL input per prep
  • Purify AAV cell pellet from 1 mL of input per prep
  • Yields highly active AAV for in vivo and in vitro experiments
  • Up to 33X sample concentration
  • Purification is based on spin column chromatography that uses Norgen’s proprietary Silicon Carbide resin separation matrix

AAV Expression Systems

Get the efficiency of transduction without viral integration with System Biosciences AAV-based expression systems—great for gene therapy research and more.

Recombinant AAV vectors are a popular go-to gene expression system for gene therapy development and gene editing in vivo because of their broad tropism, lack of associated disease, the ability to transduce both dividing and non-dividing cells, and long-term transgene expression. To help researchers take advantage of the powerful AAV system, SBI has developed a series of AAV vectors optimized for easy cloning and high titers.

Product example: pAAVK-EF1α-MCS-CMV-mRFP AAVanced™ Cloning and Expression Vector

paavk-ef1-alpha-mcs-cmv-mrfp