Antibodies for the Detection of Adeno-Associated Viruses (AAV)
AAV antibodies are suitable for the characterization of different stages of an AAV infection and are effective in the analysis of the AAV assembly process. There are two categories of AAV antibodies:
(i) AAV capsid protein antibodies that specifically react with denatured AAV capsid proteins and thereby recognize the linear epitopes of the viral capsids.
(ii) 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.
AAV Antibodies Guide
Download AAV Antibodies Guide
AAV Capsid Protein Antibodies
The AAV capsid protein antibodies recognize linear epitopes of the viral capsid proteins that consist of neighboring amino acids. These epitopes have high sequence similarity and therefore the antibodies detect the viral capsid proteins VP1, VP2 and VP3 across the serotypes. The antibodies are useful to study the expression levels of VP1, VP2 and VP3 in western blot analyses, in order to determine the protein localizations or protein-protein interactions by affinity chromatography, immunoprecipitation or immunofluorescence.
For more information on our specific AAV capsid protein antibodies, please see our VP1, VP1/VP2 and VP1/VP2/VP3 antibodies below.
Some of the following antibodies are also available in sample sizes of 10µg (lyophilized).
| anti-AAV VP1 mouse monoclonal, A1, lyophilized, purified | Cat. No. 61056 |
| anti-AAV VP1/VP2 mouse monoclonal, A69, lyophilized, purified | Cat. No. 61057 |
Binding sites and reactivity
The monoclonal antibody B1, as well as the polyclonal serum VP51 react with the three AAV capsid proteins VP1, VP2 and VP3. Both antibodies were raised against the AAV2 capsid proteins and are suitable for application in western blot due to their reactivity with linear epitopes present in the dentaured protein sequence of all three proteins. The B1 antibodies does not detect assembled AAV capsids as demonstrated in the dotplot analysis using intact and denatured AAV capsids of the serotypes AAV6 and AAV8.
The antibodies are useful for the characterization of different stages of AAV infection, the determination of the presence and identity of VP1/VP2/VP3 as well as the analysis of their expression levels and ratios during vector production.
Epitope mapping experiments (Wobus et al., 2000) identified aa726 to aa733 (C-terminus; common to all 3 VP proteins) as the specific binding region for the monoclonal B1 antibody. For more information regarding binding sites please read the publication below.
Cross-reactivity
Due to the high sequence similarity of the VP proteins derived from different AAV serotypes, the monoclonal B1 antibody shows a high cross-reactivity.
Sensitivity
PROGEN's VP1/VP2/VP3 Antibodies are highly sensitive towards AAV capsid proteins and are able to detect lowest concentrations of proteins.
The AFDye™
The use of a AFDyeTM labeled antibody enables the detection of proteins without the need of a secondary antibody which saves valuable time during processing. Theses Dyes are structurally similar to Alexa Fluor™ which is a trademark of Thermo Fisher Scientific. PROGEN offers two different AFDyes™ conjugates: AFDye™647 and AFDye™488.
AFDye™647
The AFDye™ 647 is a bright, far-red-fluorescent dye ideally suited for the 633 nm or 647 nm laser lines (excitation/emission 648/671 nm). It is structurally similar to Alexa Fluor® 647, and can be used with a common Cy5 filter set. A significant advantage to using long wavelength dyes such as Cy5 or AFDye™ 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.
AFDye™488
The AFDye™ 488 is a green emitting dye, structurally and spectrally equivalent to Alexa Fluor® 488 and therefore ideally suited for the 488 nm laser line (excitation/emission 494/517 nm). It can be used with a common FITC filter set.
| anti-AAV VP1/VP2/VP3 mouse monoclonal, B1, lyophilized, purified | Cat. No. 61058 |
| anti-AAV VP1/VP2/VP3 mouse monoclonal, B1, lyophilized, purified, sample | Cat. No. 61058S |
| anti-AAV VP1/VP2/VP3 mouse monoclonal, B1, supernatant | Cat. No. 65158 |
| anti-AAV VP1/VP2/VP3 rabbit polyconal (VP51), serum | Cat. No. 61084 |
| anti-AAV VP1/VP2/VP3 mouse monoclonal, B1, AFDyeTM 647 Conjugate | Cat. No. 61058-647 |
| anti-AAV VP1/VP2/VP3 mouse monoclonal, B1, AFDyeTM 488 Conjugate | Cat. No. 61058-488 |
AAV Particle Antibodies
The AAV particle antibodies recognize conformational epitopes of the viral capsid proteins that are only present on fully assembled capsids. These epitopes consist of distant amino acids either of the same protein or of different proteins that are in close proximity, and they might vary across the different serotypes. However, for some of the antibodies some cross-reactivity is known (for more information, please see the specific AAV particle antibodies by AAV serotype below). The AAV particle antibodies are useful for the following processes:
- to study fully assembled and intact capsids by dot blot analyses
- to quantify the intact viral capsids (empty and full) by ELISA
- to detect neutralizing antibodies in patients serum
- to determine protein-protein interaction or to purify virus preparations by affinity chromatography or immunoprecipitations
- to analyze virus particle localizations by immunofluorescence.
Some of the following antibodies are also available in sample sizes of 10µg (lyophilized).
Potential Binding Sites of the ADK1a/1b Antibody
The ADK1a antibody used for the AAV1 Titration ELISA as well as the ADK1b specifically recognize assembled AAV1 capsids, hence epitopes that result from a specific conformational assembly of the capsid proteins VP1, VP2 and VP3. In the publication cited below the contact sites and footprint residues identified for ADK1a and ADK1b are described and the results are summarized in table 1. Multiple contact sites and footprint residues have been identified, that are very likely to be part of the binding site. The amino acids of each binding site are located in different parts of the protein chains and are recognized as the epitope of the antibody only in the assembled capsid where they are in close proximity to each other and in the correct conformation.
Neutralizing activity of the ADK1 Antibody
For AAV1 the neutralization epitope has been identified and might give an additional indication about the binding residues of the neutralizing antibody ADK1 used for the corresponding AAV1 Titration ELISAs.
Find more information on the neutralizing activity of the ADK1 antibody in the following publication:
| anti-AAV1 (intact particle) mouse monoclonal, ADK1a, lyophilized, purified | Cat. No. 610150 |
| anti-AAV1 (intact particle) mouse monoclonal, ADK1a, lyophilized, purified, sample | Cat. No. 610150S |
| anti-AAV1 (intact particle) mouse monoclonal, ADK1a, Biotin Conjugate | Cat. No. 615150 |
AAV1 is also recognized by ADK8 (see AAV8).
Potential Binding Sites of the A20 Antibody
The A20 and A20R antibodies used for the AAV2 Titration ELISAs (PRATV & PRAAV2R) specifically recognize assembled AAV2 capsid, hence epitopes that result from a specific conformational assembly of the capsid proteins VP1, VP2 and VP3. Several publications cited below describe the analysis of binding sites for the A20 antibody using different techniques. Multiple amino acids have been identified, that are very likely to be part of the binding site, especially those that were identified with more than one method (green boxes). The amino acids of each binding site are located in different parts of the protein chains and are recognized as the epitope of the antibody only in the assembled capsid where they are in close proximity to each other and in the correct conformation.
Neutralizing Activity of the A20 & A20R Antibody
The A20 (Cat. No.61055) and the A20R antibody (Cat. No.610298S) were tested for neutralization using different antibody concentrations. After the HeLa cells were seeded in a 96-well plate they were incubated for 48h with the AAV2 particles containing a GFP reporter and the indicated concentration of the A20 or A20R antibody, respectively.
The microscopic analysis of the cells clearly demonstrates the neutralizing activity of the A20 and A20R antibody by decreasing GFP expression with increasing antibody concentration.
Find more information on the neutralizing activity of the A20 antibody in the following publications:
| anti-AAV2 (intact particle) mouse monoclonal, A20, lyophilized, purified | Cat. No. 61055 |
| anti-AAV2 (intact particle) mouse monoclonal, A20, lyophilized, purified, sample | Cat. No. 61055S |
anti-AAV2 (intact particle) mouse monoclonal, A20, Biotin Conjugate | Cat. No. 61555 |
| anti-AAV2 (intact particle) mouse monoclonal, A20, supernatant | Cat. No. 65155 |
| anti-AAV2 (intact particle) mouse recombinant, A20R, lyophilized, purified | Cat. No. 610298 |
| anti-AAV2 (intact particle) mouse recombinant, A20R, lyophilized, purified, sample | Cat. No. 610298S |
| anti-AAV2 (intact particle) mouse recombinant, A20R, Biotin Conjugate | Cat. No. 615298 |
AAV3 is recognized by A20 (see AAV2) and ADK8 (see AAV8).
| anti-AAV4 (intact particle) mouse monoclonal, ADK4, lyophilized, purified | Cat. No. 610147 | ||
| anti-AAV4 (intact particle) mouse monoclonal, ADK4, lyophilized, purified, sample | Cat. No. 610147S | ||
| anti-AAV4 (intact particle) mouse monoclonal, ADK4, Biotin Conjugate | Cat. No. 615147 |
Potential Binding Sites of the ADK5a/5b Antibody
The ADK5a antibody used for the AAV5 Titration ELISA as well as the ADK5b specifically recognize assembled AAV5 capsids, hence epitopes that result from a specific conformational assembly of the capsid proteins VP1, VP2 and VP3. In the publication cited below the contact sites and footprint residues identified for ADK5a and ADK5b are described and the results are summarized in table 1. Multiple contact sites and footprint residues have been identified, that are very likely to be part of the binding site. The amino acids of each binding site are located in different parts of the protein chains and are recognized as the epitope of the antibody only in the assembled capsid where they are in close proximity to each other and in the correct conformation.
Neutralizing Activity of the ADK5a & ADK5b Antibody
The ADK5a (Cat. No. 610148) and the ADK5b antibody (Cat. No. 610149) were tested for neutralization using different antibody concentrations. After the HeLa cells were seeded in a 96-well plate they were incubated for 48h with the AAV5 particles containing a GFP reporter and the indicated concentration of the ADK5a or ADK5b antibody, respectively.
The microscopic analysis of the cells clearly demonstrates the neutralizing activity of the ADK5a or ADK5b antibody by decreasing GFP expression with increasing antibody concentration.
Find more information on the neutralizing activity of the ADK5b antibody in the following publication:
| anti-AAV5 rabbit polyclonal, serum | Cat. No. 610137 |
| anti-AAV5 (intact particle) mouse monoclonal, ADK5a, lyophilized, purified | Cat. No. 610148 |
| anti-AAV5 (intact particle) mouse monoclonal, ADK5a, lyophilized, purified, sample | Cat. No. 610148S |
| anti-AAV5 (intact particle) mouse monoclonal, ADK5a, Biotin Conjugate | Cat. No. 615148 |
| anti-AAV5 (intact particle) mouse monoclonal, ADK5b, lyophilized, purified | Cat. No. 610149 |
| anti-AAV5 (intact particle) mouse monoclonal, ADK5b, lyophilized, purified, sample | Cat. No. 610149S |
Neutralizing Activity of the ADK6 Antibody
The ADK6 antibody (Cat. No. 610159) was tested for neutralization using different antibody concentrations. After the HeLa cells were seeded in a 96-well plate they were incubated for 48h with theAAV6 particles containing a GFP reporter and the indicated concentration of the ADK6 antibody.
The microscopic analysis of the cells clearly demonstrates the neutralizing activity of the ADK6 antibody by decreasing GFP expression with increasing antibody concentration.
Find more information on the neutralizing activity of the ADK6 antibody in the following publication:
| anti-AAV6 (intact particle) mouse monoclonal, ADK6, lyophilized, purified | Cat. No. 610159 | |
| anti-AAV6 (intact particle) mouse monoclonal, ADK6, lyophilized, purified, sample | Cat. No. 610159S | |
| anti-AAV6 (intact particle) mouse monoclonal, ADK6, biotin conjugate | Cat. No. 615159 |
AAV6 is also recognized by ADK1 (see AAV1).
AAV7 is recognized by ADK8 (see AAV8).
Potential Binding Sites of the ADK8 Antibody
The ADK8 antibody used for the AAV8 Titration ELISA (PRAAV8) specifically recognizes assembled AAV8 capsid, hence epitopes that result from a specific conformational assembly of the capsid proteins VP1, VP2 and VP3. The publication cited below shows the result of the Cryo-EM analysis of binding sites for the ADK8 antibody. In this publication one potential epitope has been identified which is likely to be part of the ADK8 binding site.
Neutralizing Activity of the ADK8 Antibody
The ADK8 antibody (Cat. No. 610160) was tested for neutralization using different antibody concentrations. After the HeLa cells were seeded in a 96-well plate they were incubated for 48h with theAAV8 particles containing a GFP reporter and the indicated concentration of the ADK8 antibody.
The microscopic analysis of the cells clearly demonstrates the neutralizing activity of the ADK8 antibody by decreasing GFP expression with increasing antibody concentration.
Find more information on the neutralizing activity of the ADK8 antibody in the following publications:
| anti-AAV8 (intact particle) mouse monoclonal, ADK8, lyophilized, purified | Cat. No. 610160 |
| anti-AAV8 (intact particle) mouse monoclonal, ADK8, lyophilized, purified, sample | Cat. No. 610160S |
| anti-AAV8 (intact particle) mouse monoclonal, ADK8, biotin conjugate | Cat. No. 615160 |
| anti-AAV8 (intact particle) mouse monoclonal, ADK8, supernatant | Cat. No. 651160 |
| anti-AAV8/9 (intact particle) mouse monoclonal, ADK8/9, supernatant | Cat. No. 651161 |
Neutralizing Activity of the ADK9 & ADK9-1R Antibody
The ADK9 (Cat. No. 690162) and the ADK9-1R antibody (Cat. No. 610178) were tested for neutralization using different antibody concentrations. After the HeLa cells were seeded in a 96-well plate they were incubated for 48h with the AAV9 particles containing a GFP reporter and the indicated concentration of the ADK9 or the ADK9-1R antibody, respectively.
The microscopic analysis of the cells clearly demonstrates the neutralizing activity of the ADK9 and ADK9-1R antibody by decreasing GFP expression with increasing antibody concentration.
Find more information on the neutralizing activity of the ADK9 antibody in the following publications:
Neutralization epitope of the ADK9 Antibody
For AAV9 the neutralization epitope has been identified and might give an additional indication about the binding residues of the neutralizing antibody ADK9 used for the corresponding AAV9 Titration ELISAs.
| anti-AAV-8/9 (intact particle) mouse monoclonal, ADK8/9 supernatant | Cat. No. 651161 |
| anti-AAV9 (intact particle) mouse monoclonal, ADK9, supernatant | Cat. No. 651162 |
| anti-AAV9 (intact particle) mouse monoclonal, ADK9, liquid, purified | Cat. No. 690162 |
| anti-AAV9 (intact particle) mouse monoclonal, ADK9, liquid, purified, sample | Cat. No. 690162S |
anti-AAV9 (intact particle) mouse recombinant, ADK9-1R, lyophilized, purified | Cat. No. 610178 |
anti-AAV9 (intact particle) mouse recombinant, ADK9-1R, lyophilized, purified, sample | Cat. No. 610178S |
| anti-AAV9 (intact particle) mouse monoclonal, ADK9, biotin conjugate | Cat. No. 615162 |
AAVrh10 is recognized by ADK8 (see AAV8).
AAV12 is recognized by ADK1a (see AAV1).
AAVrh74 is recognized by ADK8 (see AAV8) and ADK8/9 (see AAV8 & AAV9).
Anc80 is recognized by A20 (see AAV2) and ADK8 (see AAV8).
Anti-AAV Replicase proteins
The antibodies react with selected Rep (replicase) proteins in human AAV-infected cells.
| anti-AAV2 Replicase mouse monoclonal, 76.2, lyophilized, purified | Cat. No. 61073 |
| anti-AAV2 Replicase mouse monoclonal, 226.7, supernatant | Cat. No. 65172 |
| anti-AAV2 Replicase mouse monoclonal, 259.5, lyophilized, purified | Cat. No. 61071 |
| anti-AAV2 Replicase mouse monoclonal, 303.9, lyophylized, purified | Cat. No. 61069 |
