AAV2 and AAVX Titer Capsid ELISA Kits

Features

Multiple Serotype Coverage

• AAVX ELISA Kit detects various AAV serotypes, including but not limited to AAV1, 2, 5, 6, 8, and 9

In-House Raw Material Production

• Antibody raw materials and standards ensure high purity, specificity, consistency, and stability

Superior Kit Performance

• Wide linear range
• High precision and accuracy

Fast Detection Time

• Complete testing is achievable within 2.5 hours

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Learn more about AAV Titer Capsid ELISA Kit

• Assay Principal

  AAV Titer Capsid ELISA Kit is based on the sandwich ELISA method.

  GenScript AAV Titer Capsid Protein ELISA Kit employs two monoclonal antibodies that bind to distinct epitopes on the AAV capsid protein. When standards or samples are added to the capture plate, the capsid proteins are immobilized by the plate-bound capture antibodies. A biotin-conjugated detection antibody is then introduced, which binds to the captured capsid proteins. Streptavidin-HRP is added to interact with the biotin-conjugated antibody. After washing, TMB substrate and stop solution are applied, and the absorbance at 450 nm is measured with a microplate reader. The concentration of capsid proteins in the samples is precisely quantified using a standard curve.

  

  Figure 1. Schematic diagram of AAV Titer Capsid ELISA Kit

Validation Data

• Dilution Linearity

  Table 1a. Dilution Linearity Verification for AAVX Titer Capsid ELISA Kit - Multiple Serotypes (GenScript, L01035)

  Sample No.	Dilution Factor	Measured Conc. (Capsids/mL)	CV
  Sample A	36	2.11E+11	3%
  	72	2.18E+11
  	144	2.10E+11
  	288	2.26E+11
  Sample B	60	2.29E+11	8%
  	120	2.55E+11
  	240	2.74E+11
  	480	2.73E+11
  Sample C	50	1.10E+11	9%
  	100	1.31E+11
  	200	1.33E+11
  	400	1.19E+11
  Sample D	60	1.03E+11	9%
  	120	1.12E+11
  	240	9.39E+10
  	480	5.99E+10
  Sample E	50	9.78E+10	3%
  	100	1.03E+11
  	200	9.87E+10
  	400	5.94E+10

  Table 1b. Dilution Linearity Verification for AAV2 Titer Capsid ELISA Kit
  (GenScript, L00942)

  Sample No.	Dilution Factor	Measured Conc. (Capsids/mL)	CV
  Sample 1	120	1.26E+11	6%
  	240	1.31E+11
  	480	1.20E+11
  	960	1.39E+11
  Sample 2	130	1.37E+11	4%
  	260	1.38E+11
  	520	1.28E+11
  	1040	1.41E+11
  Sample 3	110	1.02E+11	8%
  	220	1.08E+11
  	440	1.03E+11
  	880	1.20E+11
  Sample 4	130	2.70E+11	5%
  	260	2.76E+11
  	520	2.65E+11
  	1040	2.98E+11
  Sample 5	80	2.00E+11	5%
  	160	2.19E+11
  	320	2.20E+11
  	640	2.24E+11

  The samples were diluted at different ratios and analyzed using the AAVX Titer Capsid ELISA Kit - Multiple Serotypes (GenScript, L01035) and the AAV2 Titer Capsid ELISA Kit (GenScript, L00942) to assess dilution linearity. The results showed that for the same sample, the calculated concentrations at different dilution ratios had a CV of less than 10%, indicating minimal dilution error and negligible interference from the sample matrix.

• High Precision

  Table 2a. Reproducibility Assessment for AAVX Titer Capsid ELISA Kit - Multiple Serotypes

  Batch No.	Repeats	Average Measured Conc. (Capsids/mL)	% CV	Theoretical Conc. (Capsids/mL)	Recovery Rate
  #1	10	4.03E+08	4%	4.00E+08	101%
  	10	1.24E+09	3%	1.20E+09	103%
  	10	4.70E+09	2%	5.00E+09	94%
  #2	10	4.32E+08	4%	4.00E+08	108%
  	10	1.28E+09	6%	1.20E+09	107%
  	10	4.58E+09	6%	5.00E+09	92%
  #3	10	3.94E+08	6%	4.00E+08	99%
  	10	1.24E+09	8%	1.20E+09	103%
  	10	4.50E+09	6%	5.00E+09	90%

  Table 2b. Reproducibility Assessment for AAV2 Titer Capsid ELISA Kit

  Batch No.	Repeats	Average Measured Conc. (Capsids/mL)	% CV	Theoretical Conc. (Capsids/mL)	Recovery Rate
  #1	10	1.92E+08	6%	1.80E+08	107%
  	10	4.98E+08	7%	5.00E+08	100%
  	10	1.95E+09	2%	2.00E+09	98%
  #2	10	1.83E+08	5%	1.80E+08	102%
  	10	4.85E+08	4%	5.00E+08	97%
  	10	1.95E+09	6%	2.00E+09	98%
  #3	10	1.79E+08	6%	1.80E+08	99%
  	10	4.69E+08	5%	5.00E+08	94%
  	10	1.94E+09	6%	2.00E+09	97%

  Table 3a. Inter-Batch Variability Assessment for AAVX Titer Capsid ELISA Kit - Multiple Serotypes

  Batch Amount	Repeats	Average Measured Conc. (Capsids/mL)	% CV	Theoretical Conc. (Capsids/mL)	Recovery Rate
  3	3×10	4.09E+08	6%	4.00E+08	102%
  	3×10	1.26E+09	6%	1.20E+09	105%
  	3×10	4.59E+09	5%	5.00E+09	92%

  Table 3b. Inter-Batch Variability Assessment for AAV2 Titer Capsid ELISA Kit

  Batch Amount	Repeats	Average Measured Conc. (Capsids/mL)	% CV	Theoretical Conc. (Capsids/mL)	Recovery Rate
  3	3×10	1.47E+08	9%	1.80E+08	82%
  	3×10	4.83E+08	5%	5.00E+08	97%
  	3×10	1.88E+09	6%	2.00E+09	94%

  Three different batches of the kits were used to test samples at three concentration levels, with each sample being tested in triplicate. This assessment aimed to evaluate both within-batch and inter-batch precision for the AAVX Titer Capsid ELISA Kit - Multiple Serotypes (GenScript, L01035) and the AAV2 Titer Capsid ELISA Kit (GenScript, L00942). The results indicated that the precision for both kits was less than 10% CV.

• High Recovery Rate

  Table 4a. Recovery Validation for AAVX Titer Capsid ELISA Kit - Multiple Serotypes (GenScript, L01035)

  Spiked Conc. (Capsids/mL)	Batch No.	Recovery Rate
  1.50E+08	#1	98%
  	#2	109%
  	#3	94%

  Table 4b. Recovery Validation for AAV2 Titer Capsid ELISA Kit
  (GenScript, L00942)

  Spiked Conc. (Capsids/mL)	Batch No.	Recovery Rate
  2.00E+08	#1	100%
  	#2	103%
  	#3	93%

  Three different batches of the kits were used to test spiked samples, assessing the recovery rates for the AAVX Titer Capsid ELISA Kit - Multiple Serotypes (GenScript, L01035) and the AAV2 Titer Capsid ELISA Kit (GenScript, L00942). The results indicated that the recovery rates for both kits were within the 80%-120% range.

• Correlation between L01035 and L00942 Detection
  

  Figure 2. Correlation between the AAVX Titer Capsid ELISA Kit - Multiple Serotypes (Catalog No.: L01035) and the AAV2 Titer Capsid ELISA Kit (Catalog No.: L00942) in detecting six identical AAV2 samples, with a correlation coefficient of R² = 0.9971.

• AAV2.7m8 Sample Detection Case Study
  

  Figure 3. Both the GenScript AAVX Titer Capsid ELISA Kit - Multiple Serotypes (Catalog No.: L01035) and the AAV2 Titer Capsid ELISA Kit (Catalog No.: L00942) can detect the modified AAV2.7m8.

• Detection Range of L01035 for Different Serotypes

  Table 5. Typical Assay Range for Titer Determination of Different AAV Serotypes

  Serotype	Typical assay working ranges (Capsids/mL)
  AAV1	2.73E+07 - 1.75E+09
  AAV2	1.32E+08 - 7.00E+09
  AAV5	6.30E+07 - 4.00E+09
  AAV6	3.91E+07 - 2.50E+09
  AAV8	6.27E+07 - 4.00E+09
  AAV9	1.09E+08 - 7.00E+09

  *AAV2 was tested with GenScript’s in-house produced VLPs (Virus-like Particles), while the other serotypes were tested with empty capsids from Progen.

  

  Figure 4. Standard curve for titration of different AAV serotypes

• Specificity of L00942
  

  Figure 5. Specificity of the AAV2 Titer Capsid ELISA Kit (GenScript, L00942). The kit specifically detects AAV2 samples without cross-reacting with AAV1, AAV5, AAV6, AAV8, or AAV9 samples.

FAQ

• 1. Is a positive standard provided in the kit?

  AAV2 Titer Capsid ELISA Kit (GenScript, L00942) provides AAV2 standard, which is VLP (Virus-like particle) produced by GenScript.
  The AAVX Titer Capsid ELISA Kit - Multiple Serotypes (Cat. No. L01035) only provides AAV2 standards and does not provide standards for other serotypes. Standards for other serotypes need to be established by the user.

• 2. If there are no AAV positive standards for other serotypes, can the AAVX Titer Capsid ELISA Kit - Multiple Serotypes be used?

  It is recommended to use a positive standard matching the AAV capsid in your sample to create an accurate quantitative standard curve.
  If standards for other AAV serotypes are unavailable, you can use the AAV2 standard provided with this kit to establish a standard curve and measure the sample concentration. Apply the measured concentration to the conversion formula provided to estimate the titer of the corresponding serotype. Note that this method provides a rough estimate. However, after many testing from GenScript, the correclation is kindly provided as below for references.

  Serotype	Conversion formula
  AAV1	y = 0.2498x - 3E+07
  AAV5	y = 0.8306x - 2E+07
  AAV6	y = 0.3759x - 3E+07
  AAV8	y = 0.5314x - 4E+06
  AAV9	y = 0.4569x + 9E+07

• 3. AAVX Titer Capsid ELISA Kit - Multiple Serotypes has a wide range of detection for different AAV serotypes.

  The kit has been validated for AAV serotypes 1, 2, 5, 6, 8, and 9. The typical working range for each serotype is detailed in the table below :

  Serotype	Typical assay working ranges (Capsids/mL)
  AAV1	2.73E+07 - 1.75E+09
  AAV2	1.32E+08 - 7.00E+09
  AAV5	6.30E+07 - 4.00E+09
  AAV6	3.91E+07 - 2.50E+09
  AAV8	6.27E+07 - 4.00E+09
  AAV9	1.09E+08 - 7.00E+09

  *For validation, AAV2 was tested using GenScript’s VLPs, and other serotypes were tested with empty capsids from Progen.

  The kit’s antibodies also recognize AAV2.7m8, AAVDJ, and AAVrh.1, but these serotypes have not been formally validated. While not all serotypes are validated, users can assess the kit's applicability to other serotypes according to the provided protocol.

• 4. How many samples can be tested with one test kit?

  Each kit is equipped with a pre-coated 96-well ELISA plate, which contains 8 wells x 12 strips. To ensure reliable quantification, prepare 8 standard working solutions as recommended. Testing standards and samples in duplicate is advised. Based on this setup, one kit allows for approximately 40 repeat measurements once the standard curve is applied.

• 5. Are additional wash reagents available?

  Yes, the 20× Wash Solution (GenScript, B00063) can be purchased separately and is suitable for use with this kit.

• 6. Are the measured values outside the linear range valid?

  No, values outside the linear range are invalid. Adjust the dilution factor and retest to obtain accurate results.

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