Detect viral capsid proteins VP1, VP2 and VP3

 

PROGEN's AAV capsid protein antibodies recognize linear epitopes of the viral capsid proteins VP1, VP2 and VP3. The high sequence similiarity of AAV capsid proteins allows the easy detection of proteins across serotypes.

Figure 1 shows the three capsid proteins and the corresponding antibody binding sites. As all VP proteins are encoded by the same reading frame they only differ in their N-terminal region due to the use of an alternate start codon and alternate splicing. Antibodies binding the C-terminal part, e.g. B1, are able to bind to all of the capsid proteins whereas antibodies like A1 can only bind to VP1 and A69 to VP1 and VP2.

Capsid protein antibodies can be used in western blot analyses to detect the expression levels of VP1, VP2 and VP3, as well as for protein localization studies or protein-protein interactions using chromatography, immunoprecipitation or immunofluorescence.

 

View AAV Capsid Protein Antibodies

Binding Sites and Reactivity

Capsid protein antibodies were raised in mice against AAV2 capsid proteins and are suitable for all applications detecting linear epitopes of the denatured VP protein, such as western blot analysis.

Serotype Antibody Residues* Binding Site
AAV2 A1 123-KRVLEPLGL-131 Binding site is located at the N-terminus of the VP1 protein. Detection of VP1 only.
AAV2 A69 171-LNFGQTGDADSV-182 Binding site is located at the N-terminus of the VP2 protein. Detection of VP1 and VP2.
AAV2  B1 726-IGTRYLTR-733 Binding site is located at the C-terminus common to all three VP proteins. Detection of VP1, VP2 and VP3.
*The amino acid position is with regards to the VP1 sequence. 

Epitope mapping experiments (Wobus et al., 2000) identified aa123 to aa131 (N-terminus;) as the specific binding region for the monoclonal A1 antibody. 

Wobus, C. E. et al. Monoclonal antibodies against the adeno-associated virus type 2 (AAV-2) capsid: epitope mapping and identification of capsid domains involved in AAV-2-cell interaction and neutralization of AAV-2 infection. J. Virol. 74, 9281–93 (2000).

 

View AAV Capsid Protein Antibodies

Cross-reactivity

Antibody AAV1 AAV2 AAV3 AAV4 AAV5 AAV6 AAV7 AAV8 AAV9 AAVrh10 AAV11 AAV12 AAVDJ
A1 x x x   x x x x x   x x x
A69   x         na       na na x
B1 x x x   x x x x x x     x
VP51 na x na na x na na na na na na na na

 

x - Reactivity
na - Not analyzed
Empty field - no reactivity

A1

Reacts with VP1 proteins from all common serotypes AAV1-12. In immunoprecipitation a reaction with a non-AAV derived protein can occasionally be seen.

 

View AAV Capsid Protein Antibodies

B1

The B1 antibody shows high cross-reactivity with most of the tested AAV serotypes due to high sequence similarities of VP proteins derived from different AAVs. 

Using AFDyes

Using an AFDye™ labeled antibody allows you to detect your proteins of interest without using a secondary antibody. This saves you valuable time as you can also simultaneously detect different proteins in the same sample by using differently labelled antibodies e.g.  AFDye™ 647 and AFDye™ 488.

PROGEN offers two different AFDye™ conjugates for the B1 antibody: AFDye™ 647 and AFDye™ 488.

AFDyeTM 647

Appearance Bright deep red fluorescent
Excitation 648 nm
Emission 671 nm
Filterset E.g. Cy5
Spectrally similar Dyes Alexa Fluor® 647
Advantage

Low autofluorescence due to long wavelength

 

This western blot shows the analysis of different AAV serotpyes using our anti-AAV VP1/VP2/VP3 AFDyeTM 647 antibody.

 

View AFDye 647

AFDyeTM 488

Appearance Bright green fluorescent
Excitation 494 nm
Emission 517 nm
Filterset E.g. FITC
Spectrally similar Dyes Alexa Fluor® 488


This western blot shows the analysis of different AAV serotypes using our anti-AAV VP1/VP2/VP3 AFDyeTM 488 antibody.

View AFDye 488

AAV Antibodies FAQs

Yes, our AAV particle antibodies detect conformational epitopes only present on assembled capsids.

You can find all AAV particle antibodies here.

In contrast, AAV capsid protein antibodies recognize linear epitopes present on denatured VP proteins.

You can find all AAV capsid protein antibodies here.

Yes, PROGEN offers a selection of AAV antibodies in 10 µg sample sizes. You can find all available AAV antibodies here: AAV Antibodies

PROGEN offers a wide range of AAV antibodies either detecting a linear epitope (AAV capsid protein antibodies), a conformational epitope (AAV particle antibodies) or a AAV replicase.

These antibodies are provided in different quantities and formats (e.g. purified and supernatant). A selection of AAV antibodies are also available in 10 µg sample sizes. You can find all available AAV antibodies here: AAV Antibodies.

Anti-Adeno-Associated Virus Capsid Proteins Antibodies

For more information on the antibody binding epitopes of the capsid protein antibodies A1, B1 and A69 see the table below summarizing the published data from the following publication on the specific antibody binding sites:

Table Binding Sites of the AAV capsid protein antibodies

Wobus, C. E. et al. Monoclonal antibodies against the adeno-associated virus type 2 (AAV-2) capsid: epitope mapping and identification of capsid domains involved in AAV-2-cell interaction and neutralization of AAV-2 infection. J. Virol. 74, 9281–93 (2000).

Anti-Adeno-Associated Virus Particle Antibodies

For more information on the antibody binding epitopes on assembled AAV1, 2, 5, 8 and 9 capsids, see the tables below summarizing the published data from the following publications on the specific antibody binding sites:

summary binding epitopes


EM structures of assembled AAV1, 2, 4, 5, 6, and 8 capsids with Fab-fragments are available in the following publications: