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AAV Antibodies and Titration ELISA

Adeno-Associated Virus (AAV) is a nonpathogenic virus that is widely used as vector in gene therapy. Creative Diagnostics currently provide full range of adeno-associated virus (AAV) antibodies that specifically recognize intact virus capsids as well as antibodies specific to AA-viral subunits and replicases. Titration ELISA kits for various AAV subtype is also available.

Adeno-Associated Virus (AAV) Related Antibodies

Cat. No. Product Name Description
DMAB6351 Anti-AAV1 (intact particle) monoclonal antibody, Clone BEL2b [Biotin] Specifically detects AAV1 intact virus particles, both empty and full capsids. DMAB6351 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. Not suitable for Immunoblotting. In immunofluorescence, a week cross-reactivity was observed with AAV6. No reaction with AAV type 2, 3, 4, 5.
DMAB6350 Anti-AAV1 (intact particle) monoclonal antibody, Clone BEL2b Specifically detects AAV1 intact virus particles, both empty and full capsids. DMAB6350 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. Not suitable for Immunoblotting. In immunofluorescence and dot plot analysis, a week cross-reactivity was observed with AAV6 and AAV12. No reaction with other AAV types.
CABT-B9061 Anti-AAV2 (intact particle) monoclonal antibody, clone B31 [Biotin] Specifically detects intact virus particles, both empty and full capsids. With AAV2 and AAV3 CABT-B9061 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. Not for WB.
CABT-B9062 Anti-AAV2 (intact particle) monoclonal antibody, clone B31 Specifically detects intact virus particles, both empty and full capsids. With AAV2 and AAV3 CABT-B9061 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. Not for WB.
DMAB6346 Anti-AAV2 Replicase monoclonal antibody, Clone A227.8 Recognizes AAV replicase proteins Rep 78, Rep 68, Rep 52 and Rep 40 in AAV-infected cells. In Immunoblotting prominant reaction with Rep 78 and Rep 68, only faint reaction with Rep 52 and Rep 40.
DMAB6345 Anti-AAV2 Replicase monoclonal antibody, Clone A304.10 Recognizes AAV replicase proteins Rep 78, Rep 68, Rep 52 and Rep 40 in human AAV-2-infected cells. Cross-reactivity with AAV serotypes 1, 3, 4, 5, 6, 7, 8, 9 and 10 (4,11 and 12 were negative). A weak cross-reactivity is seen in some human cell lines.
DMAB6347 Anti-AAV2 Replicase monoclonal antibody, Clone A77.4 Recognizes AAV replicase proteins Rep78 and Rep52 in human AAV2-infected cells, no reaction with Rep68 and Rep40. Weak cross-reaction with AAV-serotype 3.
DMAB6348 Anti-AAV2 VP1 monoclonal antibody, Clone B2 DMAB6348 reacts with VP1 of AAV-2; weak cross-reaction with serotypes 1, 3, 4, 5, 6. In immunoprecipitation, an occasional reaction with a non-AAV-derived protein is found.
DMAB6349 Anti-AAV2 VP1/VP2 monoclonal antibody, Clone B610 Recognizes AAV-2 viral capsid proteins VP1 and VP2, which are highly enriched in the nucleus. Weak cross-reaction with serogroups 1, 3, and 6.
DPAB2423 Anti-AAV2 VP1/VP2/VP3 polyclonal antibody Reacts with VP1, VP2 and VP3. Raised against recombinant AAV-2 capsid proteins.
CABT-B9063 Anti-AAV4 (intact particle) monoclonal antibody, clone BEL5 [Biotin] Specifically detects AAV4 intact virus particles, both empty and full capsids. CABT-B9063 recognizes a conformational epitope of assembled capsids not present in denatured and native unassembled capsid proteins. No cross-reaction with other AAV serotypes.
DMAB6352 Anti-AAV4 (intact particle) monoclonal antibody, Clone BEL5 Specifically detects AAV4 intact virus particles, both empty and full capsids. DMAB6352 recognizes a conformational epitope of assembled capsids not present in denatured and native unassembled capsid proteins. No cross-reaction with other AAV serotypes.
DMAB6355 Anti-AAV5 (intact particle) monoclonal antibody, Clone BEL6b [Biotin] Specifically detects intact virus particles, both empty and full capsids. DMAB6355 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. No cross-reaction with other AAV serotypes. The antibody is not suitable for Immunoblotting.
DMAB6353 Anti-AAV5 (intact particle) monoclonal antibody, Clone BEL6b Specifically detects intact virus particles, both empty and full capsids. DMAB6353 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. No cross-reaction with other AAV serotypes. The antibody is not suitable for Immunoblotting.
DMAB6354 Anti-AAV5 (intact particle) monoclonal antibody, Clone BEL6c Specifically detects intact virus particles, both empty and full capsids. DMAB6353 recognizes a conformational epitope of assembled capsids, not present in denatured and native unassembled capsid proteins. No cross-reaction with other AAV serotypes. The antibody is not suitable for Immunoblotting.
DPAB2424 Anti-AAV5 polyclonal antiserum Raised against recombinant AAV5 capsid proteins. The antibody reacts with virus capsid proteins VP1, VP2, VP3 of AAV5; no cross-reaction with AAV types 1 - 4.
CABT-B9064 Anti-AAV6 (intact particle) monoclonal antibody, clone BEL7 Useful for analysis of the AAV assembly process. CABT-B9064 specifically reacts with intact adeno-associated virus particles, empty and full capsids. Recognizes a conformational epitope of assembled capsids. Specific for AAV-6. No cross-reaction with AAV type 1, 2, 3, 4, 5, 7, 8, 9, 10, 11, and 12 was observed. Not for use in WB.
CABT-B9065 Anti-AAV8 (intact particle) monoclonal antibody, clone BEL9 For characterization of different stages of infection and very useful for analysis of the AAV assembly process. CABT-B9065 specifically reacts with intact adeno-associated virus particles, empty and full capsids. Recognizes a conformational epitope of assembled capsids. Not for use in immunoblotting.
CABT-B9066 Anti-AAV8/9 (intact particle) monoclonal antibody, clone BEL9/0 For characterization of different stages of infection and very useful for analysis of the AAV assembly process. CABT-B9066 specifically reacts with intact adeno-associated virus particles, empty and full capsids. Recognizes a conformational epitope of assembled capsids. The antibody cannot be used for immunoblotting using denaturing conditions.
DPAB-AV01 Anti-AAV9 (intact particles) monoclonal antibody, clone BEL0 Useful for analysis of the AAV assembly process. DPAB-AV01 specifically reacts with intact adeno-associated virus particles, empty and full capsids. Recognizes a conformational epitope of assembled capsids. Not for use in WB.

Adeno-Associated Virus (AAV) Titration ELISA


What Is AAV?

The first human adeno-associated virus (AAV) was discovered in 1965, as a contaminant of adenovirus (Ad) preparations. AAV is one of the smallest viruses with a non-enveloped icosahedral capsid of approximately 22 nm. Despite the high seroprevalence of AAV in the human population (approximately 80% of humans are seropositive for AAV2) the virus has not been linked to any human illness.

Gene therapy vectors using AAV can infect both dividing and quiescent cells and persist in an extrachromosomal state without integrating into the genome of the host cell, although in the native virus some integration of virally carried genes into the host genome does occur. These features make AAV a very attractive candidate for creating viral vectors for gene therapy, and for the creation of isogenic human disease models.

AAV Antibodies and Titration ELISA AAV Genome Structure, DNA Replication, and Virus Assembly

AAV has a linear single-stranded DNA (ssDNA) genome of approximately 4.7-kilobases (kb), with two 145 nucleotide-long inverted terminal repeats (ITR) at the termini. The virus does not encode a polymerase and therefore relies on cellular polymerases for genome replication. The ITRs flank the two viral genes – rep (replication) an”yd cap (capsid), encoding non-structural and structural proteins, respectively.

The cap gene, through alternative splicing and initiation of translation, gives rise to three capsid proteins, VP1 (virion protein 1), VP2 and VP3, with molecular weight of 87, 72 and 62 kDa, respectively. These capsid proteins assemble into a near-spherical protein shell of 60 subunits. The rep gene, through the use of two promoters and alternative splicing, encodes four regulatory proteins that are dubbed Rep78, Rep68, Rep52 and Rep40. These proteins are involved in AAV genome replication.

Adeno-associated Virus Vectorology

Historically, most recombinant AAV (rAAV) vectors have been based on serotype 2 (AAV2) that constitutes the prototype of the genus. Many approaches have been taken to improve AAV vector production, but two strategies differing in principles are now the most widely used, depending on how the adenovirus helper factors are provided.

One method relies on wild-type adenovirus infection into the cell lines that stably harbor AAV rep/cap genes, as well as the AAV vector DNA. The other method, the helper-free method, is based on the adenovirus-free transient transfection of all elements that are required for AAV production in host cells such as HEK293 cells. It involves the co-transfection of AAV production cells with 3 plasmids: (1) an AAV2 ITR-containing plasmid carrying the gene of interest; (2), a plasmid that carries the AAV2 Rep-Cap; and (3), a plasmid that provides the helper genes isolated from adenovirus.

AAV Antibodies and Titration ELISA

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