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Simple Plex AAV2 Cartridge

R&D Systems | Catalog # SPCKB-OT-005905

Simple Plex AAV2 assay kit contains cartridge, sample diluent SD19, and wash buffer. Part # starting with ST01 ships in 2-3 days.
R&D Systems

Key Product Details

Assay Length

90 minutes

Sample Type & Volume Required Per Well*

Bioprocess Samples (25 µL)

Sensitivity

0.9 106 capsids/mL

Assay Range

3.41-13000 106 capsids/mL

Simple Plex AAV2 Cartridge Summary

Simple Plex assay for the detection of Adeno-Associated Virus type 2 (AAV2) intact capsids in bioprocess samples. This assay uses PROGEN's AAV2 (A20R) antibody.
  • Sensitivity: 0.9 x 106 capsids/mL
  • Range: 3.41 x 106 - 1.3 x 1010 capsids/mL

*The Sample Volume represented is based on the amount of sample incorporated into the reaction after taking into account the assay's minimum required dilution for a given matrix. Serial dilution may be necessary to achieve some of the final sample volumes represented.

View all AAV2 Simple Plex »

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Product Specifications

Assay Type

Automated sandwich ELISA

Format

Microfluidic cartridge containing three Glass Nano Reactors (GNR)

Species

Viral

Specificity

This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Each control was tested 16 times in one assay.

Inter-Assay Precision (Precision between assays) Replicates of each control were tested in multiple assays performed by at least three technicians using two lots of reagents.

Bioprocess Samples

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 1 2
n 16 16 58 56
Mean (10<sup>6</sup> capsids/mL) 40 2103 42.2 2045
Standard Deviation 3.5 55.3 4.7 158
CV% 8.8 2.6 11.2 7.7

Recovery for Simple Plex AAV2 Cartridge

Recovery was not necessary as AAV2 shows natural linearity data from a neat sample to a 1:16 dilution.

Linearity

Samples containing and/or spiked with high concentrations of AAV2 were serially diluted with Sample Diluent to produce samples within the dynamic range of the assay.

Scientific Data Images for Simple Plex AAV2 Cartridge

AAV2 Simple Plex Assay

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: AAV2

Adeno-associated virus (AAV) is a small, non-enveloped single-stranded DNA virus belonging to the Pavoviridae family in the genus Dependoparvovirus, which requires a helper virus in order to replicate (1-3). The AAV genome consists of a rep and cap gene which are flanked by inverted terminal repeats (ITRs) (1-4). The rep gene encodes for AAV's non-structural proteins that function in replication, packaging, and integration (2-4). The cap gene encodes the structural proteins (VP1, VP2, VP3) that form the viral capsid made up of 60 protein subunits (2-4). Recombinant AAV vectors are generated by replacing the rep and cap genes and inserting a promoter, followed by a transgene, and then a poly-A (pA) tail, flanked by ITRs (3-4). Recombinant vectors are then packaged by providing the rep and cap genes in trans and helper genes for AAV replication (3). AAV vectors have been studied for their potential in human gene therapy and are considered a safe platform given their low immunogenicity and that they are not associated with any known diseases (1-4). Additionally, recombinant AAV vectors are typically non-integrating (1). There are 13 identified human and non-human primate AAV serotypes and over 100 natural AAV variants (2-4). The AAV serotypes are further classified into six clades (A-F) and two clonal isolates (AAV4 and AAV5) according to the VP1 amino acid (aa) sequence (3). These AAV serotypes share ~65-99% sequence identity and ~95-99% structural identity (3). Research has shown that the different AAV serotypes have varying tissue tropism and transduction efficiency based on tissue origin (2). For instance, AAV8 and AAV9 are optimal for transduction in the liver, while AAV2 is ideal for the kidney (2). AAV2 is the best characterized and most commonly used AAV serotype in preclinical studies and clinical trials (4). AAV vectors have been employed in a number oncology models for their ability to carry and deliver a variety of factors including anti-angiogenic genes, suicide genes, tumor suppressors, cytokines, and monoclonal antibodies (4). Advances in engineering AAV vectors and combining AAV gene delivery with other treatment modalities, like chemotherapeutics, is promising for the future of human gene therapy (2-4).

References

1. Zengel J, Carette JE. Structural and cellular biology of adeno-associated virus attachment and entry. Adv Virus Res. 2020;106:39-84. https://doi.org/10.1016/bs.aivir.2020.01.002

2. Wu Z, Asokan A, Samulski RJ. Adeno-associated virus serotypes: vector toolkit for human gene therapy. Mol Ther. 2006;14(3):316-327. https://doi.org/10.1016/j.ymthe.2006.05.009

3. Drouin LM, Agbandje-McKenna M. Adeno-associated virus structural biology as a tool in vector development. Future Virol. 2013;8(12):1183-1199. https://doi.org/10.2217/fvl.13.112

4. Santiago-Ortiz JL, Schaffer DV. Adeno-associated virus (AAV) vectors in cancer gene therapy. J Control Release. 2016;240:287-301. https://doi.org/10.1016/j.jconrel.2016.01.001

Long Name

Adeno-associated Virus 2

Alternate Names

Adeno-associated Virus 2, capsid, Coat protein, major coat protein

Additional AAV2 Products

Product Documents for Simple Plex AAV2 Cartridge

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Certificate of Analysis

To download a Certificate of Analysis, please enter your Simple Plex catalog number and lot number in the search boxes below.

Product Specific Notices for Simple Plex AAV2 Cartridge

For research use only

Citations for Simple Plex AAV2 Cartridge

View all citations for the Simple Plex platform on Bio-Techne.com.

Please visit our Instrument Citation Database on Bio-Techne.com to search our collection of scientific articles that feature our instruments.

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FAQs for Simple Plex AAV2 Cartridge

Showing  1 - 5 of 6 FAQs Showing All

  • Q: Do the AAV assays recognize both natural and recombinant AAV?

    A: Yes, the AAV assays detect both natural and recombinant AAV. 

  • Q: Do the AAV assays recognize fully-assembled capsids?

    A: Yes, these assays detect fully-assembled capsids. Non-assembled protein or partially assembled capsids will not be detected.

  • Q: Do the Simple Plex AAV assays distinguish between full and empty capsids?

    A: No, the AAV assays do not distinguish between full and empty capsids. Complementary methods are required to distinguish full and empty capsids.

  • Q: Does the AAV2 assay recognize other AAV serotypes?

    A: This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

  • Q: What diluent is provided with the AAV assays?

    A: The AAV assays are supplied with SD19 concentrate. The concentrate should be diluted 1:5 in deionized or dilstilled water prior to use. 

  • Q: What is the minimum required dilution (MRD) for bio-process samples?

    A: All samples require a minimum 2-fold dilution for use in Simple Plex assays, but further dilution may be needed for optimal performace. An appropriate dilution factor for each process matrix should be determined experimentally by assessment of sample linearity and spike recovery. 

  • Q: Do the AAV assays recognize both natural and recombinant AAV?

    A: Yes, the AAV assays detect both natural and recombinant AAV. 

  • Q: Do the AAV assays recognize fully-assembled capsids?

    A: Yes, these assays detect fully-assembled capsids. Non-assembled protein or partially assembled capsids will not be detected.

  • Q: Do the Simple Plex AAV assays distinguish between full and empty capsids?

    A: No, the AAV assays do not distinguish between full and empty capsids. Complementary methods are required to distinguish full and empty capsids.

  • Q: Does the AAV2 assay recognize other AAV serotypes?

    A: This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

  • Q: What diluent is provided with the AAV assays?

    A: The AAV assays are supplied with SD19 concentrate. The concentrate should be diluted 1:5 in deionized or dilstilled water prior to use. 

  • Q: What is the minimum required dilution (MRD) for bio-process samples?

    A: All samples require a minimum 2-fold dilution for use in Simple Plex assays, but further dilution may be needed for optimal performace. An appropriate dilution factor for each process matrix should be determined experimentally by assessment of sample linearity and spike recovery. 

  • Q: Do the AAV assays recognize both natural and recombinant AAV?

    A: Yes, the AAV assays detect both natural and recombinant AAV. 

  • Q: Do the AAV assays recognize fully-assembled capsids?

    A: Yes, these assays detect fully-assembled capsids. Non-assembled protein or partially assembled capsids will not be detected.

  • Q: Do the Simple Plex AAV assays distinguish between full and empty capsids?

    A: No, the AAV assays do not distinguish between full and empty capsids. Complementary methods are required to distinguish full and empty capsids.

  • Q: Does the AAV2 assay recognize other AAV serotypes?

    A: This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

  • Q: What diluent is provided with the AAV assays?

    A: The AAV assays are supplied with SD19 concentrate. The concentrate should be diluted 1:5 in deionized or dilstilled water prior to use. 

  • Q: What is the minimum required dilution (MRD) for bio-process samples?

    A: All samples require a minimum 2-fold dilution for use in Simple Plex assays, but further dilution may be needed for optimal performace. An appropriate dilution factor for each process matrix should be determined experimentally by assessment of sample linearity and spike recovery. 

  • Q: Do the AAV assays recognize both natural and recombinant AAV?

    A: Yes, the AAV assays detect both natural and recombinant AAV. 

  • Q: Do the AAV assays recognize fully-assembled capsids?

    A: Yes, these assays detect fully-assembled capsids. Non-assembled protein or partially assembled capsids will not be detected.

  • Q: Do the Simple Plex AAV assays distinguish between full and empty capsids?

    A: No, the AAV assays do not distinguish between full and empty capsids. Complementary methods are required to distinguish full and empty capsids.

  • Q: Does the AAV2 assay recognize other AAV serotypes?

    A: This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

  • Q: What diluent is provided with the AAV assays?

    A: The AAV assays are supplied with SD19 concentrate. The concentrate should be diluted 1:5 in deionized or dilstilled water prior to use. 

  • Q: What is the minimum required dilution (MRD) for bio-process samples?

    A: All samples require a minimum 2-fold dilution for use in Simple Plex assays, but further dilution may be needed for optimal performace. An appropriate dilution factor for each process matrix should be determined experimentally by assessment of sample linearity and spike recovery. 

  • Q: Do the AAV assays recognize both natural and recombinant AAV?

    A: Yes, the AAV assays detect both natural and recombinant AAV. 

  • Q: Do the AAV assays recognize fully-assembled capsids?

    A: Yes, these assays detect fully-assembled capsids. Non-assembled protein or partially assembled capsids will not be detected.

  • Q: Do the Simple Plex AAV assays distinguish between full and empty capsids?

    A: No, the AAV assays do not distinguish between full and empty capsids. Complementary methods are required to distinguish full and empty capsids.

  • Q: Does the AAV2 assay recognize other AAV serotypes?

    A: This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

  • Q: What diluent is provided with the AAV assays?

    A: The AAV assays are supplied with SD19 concentrate. The concentrate should be diluted 1:5 in deionized or dilstilled water prior to use. 

  • Q: What is the minimum required dilution (MRD) for bio-process samples?

    A: All samples require a minimum 2-fold dilution for use in Simple Plex assays, but further dilution may be needed for optimal performace. An appropriate dilution factor for each process matrix should be determined experimentally by assessment of sample linearity and spike recovery. 

  • Q: Do the AAV assays recognize both natural and recombinant AAV?

    A: Yes, the AAV assays detect both natural and recombinant AAV. 

  • Q: Do the AAV assays recognize fully-assembled capsids?

    A: Yes, these assays detect fully-assembled capsids. Non-assembled protein or partially assembled capsids will not be detected.

  • Q: Do the Simple Plex AAV assays distinguish between full and empty capsids?

    A: No, the AAV assays do not distinguish between full and empty capsids. Complementary methods are required to distinguish full and empty capsids.

  • Q: Does the AAV2 assay recognize other AAV serotypes?

    A: This assay recognizes intact AAV2 capsids. The AAV2 antibody (A20R) used in this assay cross-reacts with AAV3.

  • Q: What diluent is provided with the AAV assays?

    A: The AAV assays are supplied with SD19 concentrate. The concentrate should be diluted 1:5 in deionized or dilstilled water prior to use. 

  • Q: What is the minimum required dilution (MRD) for bio-process samples?

    A: All samples require a minimum 2-fold dilution for use in Simple Plex assays, but further dilution may be needed for optimal performace. An appropriate dilution factor for each process matrix should be determined experimentally by assessment of sample linearity and spike recovery. 

Showing  1 - 5 of 6 FAQs Showing All