Figure 1. Schematic diagram of Lentivirus Titer p24 ELISA Kit
Lentivirus Titer p24 ELISA Kit Pro can remove free p24 and only detect p24 protein on lentivirus capsid
Fewer dilutions with linear range from 31.25 to 2,000 pg/ml
The complete titration can be finished within 3 hours
With Intra-assay CV ≤ 10% and Inter-assay CV ≤ 15%
Ideal for the entire process of lentivirus vector production, capable of testing samples at various stages such as plasmid transfection, clarification filtration, chromatographic purification, and final product release
High stability p24 protein standard with scientific calibration
Flexible format, only use the amount as it’s needed
Using the easy-to-detect ELISA method, the values can be read through a microplate reader
Cat. No. | Product Name | Quantity | Price | Order |
---|---|---|---|---|
L00938
|
Lentivirus Titer p24 ELISA Kit |
|
$498.00 96 Tests |
|
L00974
|
Lentivirus Titer p24 ELISA Kit Pro |
|
$548.00 96 Tests |
|
Figure 1. Schematic diagram of Lentivirus Titer p24 ELISA Kit
Figure 2. Detection steps of L00938 and L00974
Effectively reduce interference from free p24
Figure 3. Different concentrations of p24 protein were added to the diluent buffer, the results show that Lentivirus Titer p24 ELISA Kit Pro (GenScript, L00974) barely detects free p24 added in the diluent.
Specific detection of p24 on lentiviral particles
Figure 4. Different concentrations of p24 protein were added to the purified lentivirus samples, the results show that the p24 protein concentration detected by the Lentivirus Titer p24 ELISA Kit Pro (GenScript, L00974) is consistent with the actual p24 concentration on the lentivirus particles.
Recovery is a fundamental biophysical characteristic to evaluate immunoassay. Ideally, the recovery rate should be ultimately close to 100%. 100% recovery means there is no interference from the diluent or matrix in the assay. Normally, an 80% to 120% recovery rate can be regarded as there being relatively slight interference from the diluent or matrix. From the assay in triplicates, the recovery rates of GenScript Lentivirus Titer p24 ELISA kits are all in the range of 80% to 120%, which indicates lentivirus titration with this ELISA kit is accurate and reproducible.
Table 1. Recovery rate of Lentivirus Titer p24 ELSIA kit in triplicates
Batch No. | Repeats | Average Conc. pg/mL | Recovery Rate |
---|---|---|---|
#1 | 3 | 1945.30 | 86% |
#2 | 3 | 1787.64 | 108% |
#3 | 3 | 1752.76 | 90% |
A. High intra-assay precision
The following report includes data from a representative intra-assay precision validation. Samples at low, medium, and high concentration levels were tested using GenScript p24 ELISA kits. Each sample was tested 10 times, and the coefficient of variation (CV) was calculated for each sample.
Table 2. Intra-assay CV calculation with three-batch Lentivirus Titer p24 ELISA Kit
Batch No. | Repeats | Average Conc. pg/mL | %CV |
---|---|---|---|
#1 | 10 | 215.55 | 3% |
#1 | 10 | 829.45 | 4% |
#1 | 10 | 1537.42 | 4% |
#2 | 10 | 209.60 | 5% |
#2 | 10 | 792.76 | 2% |
#2 | 10 | 1431.42 | 3% |
#3 | 10 | 188.09 | 6% |
#3 | 10 | 759.99 | 4% |
#3 | 10 | 1377.71 | 3% |
B. High inter-assay precision
The following report includes data from a representative inter-assay precision validation. Samples at low, medium, and high concentration levels were tested using GenScript p24 ELISA kits. Each sample was tested 10 times, and the coefficient of variation (CV) was calculated for each sample.
Table 3. Inter-assay CV calculation with three-batch Lentivirus Titer p24 ELISA Kit
Batch Amount | Repeats | Average Conc. pg/mL | %CV |
---|---|---|---|
3 | 3x10 | 204.41 | 7% |
3 | 3x10 | 794.07 | 5% |
3 | 3x10 | 1448.85 | 6% |
Utilizing p24 ELISA kit for lentivirus titration during lentivirus purification and production can greatly benefit lentivirus purification process control and optimization. The outstanding characteristics of GenScript Lentivirus Titer p24 ELISA kit that outshines the competitors are its extremely wide linear range from 31.25 to 2,000 pg/ml and short incubation time. With the advantage of a wide linear range, fewer dilutions of the sample would be needed to get the sample titer into the linear range. Much fewer errors that might be caused by dilution can be avoided.
Table 4a. Lentivirus titration during lentivirus purification and production using GenScript Lentivirus Titer p24 ELISA Kit
Process | OD Value | Testing p24 Conc.(pg/mL) | Dilution Factor | Actual p24 Conc. (LP/mL) | CV % |
---|---|---|---|---|---|
Supernatant | 3.151 | 2445.918 | 100 | Not Valid | / |
1.225 | 444.343 | 3000 | 1.67E+10 | ||
Nucleic Acid Removal | 3.12 | 2369.055 | 100 | Not Valid | / |
1.242 | 451.997 | 3000 | 1.69E+10 | ||
Centrifugation | 3.103 | 2328.92 | 100 | Not Valid | / |
0.94 | 321.172 | 3000 | 1.20E+10 | ||
Clarification and Filtration | 3.136 | 2405.64 | 100 | Not Valid | / |
0.926 | 315.765 | 3000 | 1.18E+10 | ||
Ultrafiltration and Diafiltration | 2.183 | 1040.264 | 3000 | 3.90E+10 | 3 |
0.152 | 36.234 | 90000 | 4.08E+10 | ||
Clarification and Filtration | 1.994 | 890.168 | 3000 | 3.34E+10 | 6 |
0.142 | 32.441 | 90000 | 3.65E+10 | ||
Gel Filtration | 1.496 | 578.496 | 3000 | 2.17E+10 | / |
0.106 | 18.902 | 90000 | Not Valid | ||
Ultrafiltration and Diafiltration | 2.623 | 1501.37 | 3000 | 5.63E+10 | 3 |
0.185 | 47.906 | 90000 | 5.39E+10 | ||
Final Product | 3.155 | 2459.286 | 3000 | Not Valid | / |
0.799 | 265.916 | 90000 | 2.99E+11 |
Table 4b. Lentivirus titration during lentivirus purification and production using p24 ELISA Kit from other vendor
Process | OD Value | Testing p24 Conc.(pg/mL) | Dilution Factor | Actual p24 Conc. (LP/mL) | CV % |
---|---|---|---|---|---|
Supernatant | 86.687 | 1.223 | 12000 | 1.30E+10 | / |
291.899 | 3.143 | 2000 | Not Valid | ||
Nucleic Acid Removal | 100.254 | 1.375 | 12000 | 1.50E+10 | / |
282.872 | 3.073 | 2000 | Not Valid | ||
Centrifugation | 73.79 | 1.073 | 12000 | 1.11E+10 | / |
275.974 | 3.019 | 2000 | Not Valid | ||
Clarification and Filtration | 69.953 | 1.028 | 12000 | 1.05E+10 | / |
279.996 | 3.051 | 2000 | Not Valid | ||
Ultrafiltration and Diafiltration | 234.108 | 2.675 | 10000 | Not Valid | / |
39.453 | 0.653 | 60000 | 2.96E+10 | ||
Clarification and Filtration | 212.127 | 2.484 | 10000 | Not Valid | / |
36.317 | 0.612 | 60000 | 2.72E+10 | ||
Gel Filtration | 165.17 | 2.048 | 10000 | 2.06E+10 | 5 |
29.342 | 0.522 | 60000 | 2.20E+10 | ||
Ultrafiltration and Diafiltration | 275.001 | 3.011 | 10000 | Not Valid | / |
69.346 | 1.021 | 60000 | 5.20E+10 | ||
Final Product | 38.46 | 0.64 | 480000 | 2.31E+11 | / |
244.786 | 2.765 | 80000 | Not Valid |
*The data highlighted in red were out of the linear range of the ELISA kit used.
During the process development for the lentivirus purification, 18 samples from 9 processes were collected and titrated. The experimental results found that: 1. The dilution factor of GenScript Lentivirus Titer p24 ELISA Kit is lower than that of the competing p24 Detection Kit. 2. In the process of diluting the lentivirus into the linear range of p24 ELISA kits, the success rate of 18 samples using the GenScript Lentivirus Titer p24 ELISA Kit with only one dilution was 67%, while the success rate was 56% when using the competing p24 ELISA kit.
There are two ways to express virus titration: functional (infectious) titration measured by transduction or physical titration measured by virus particles. Physical titration measures how much virus is present in a sample, typically calculated based on the level of proteins (such as p24). Functional titration measures how many viruses can infect cells, usually 100-1000 times lower than physical titration.
For Lentivirus Titer p24 ELISA Kit (GenScript, L00938), the diluted 1× Wash Solution from the kit can be used for sample dilution, with the same effect. For Lentivirus Titer p24 ELISA Kit Pro (GenScript, L00974), during the separation of free p24, it is not necessary to dilute the lentivirus supernatant sample. If dilution is needed, use cell culture medium (10% FBS). In the subsequent ELISA testing stage, the diluted 1× Wash Solution from the kit can be used for dilution. If using sample or formulation buffer as the diluent for standards and samples, it is necessary to test the recovery rate through pre-experiments to determine whether it meets expectations. Other diluents such as ultrapure water, DMEM without added FBS, cannot guarantee normal output results. If you need support for validation, you can contact us.
Lentivirus Titer p24 ELISA Kit Pro (GenScript, L00974), only detects p24 protein on the lentivirus capsid, not interfered by free p24 protein, and it takes about 3 hours to obtain results. Lentivirus Titer p24 ELISA Kit (GenScript, L00938) detects total p24 protein, including free p24 protein and p24 protein on the lentiviral capsid, and it takes about 2.5 hours to obtain results.
Both L00938 and L00974 don’t contain any human-derived materials and have low biological risk.
GenScript Lentivirus Titer p24 ELISA kits have a pre-coated 96-well microplate, which contains 8 wells x 12 strips. We recommend diluting the standard curve gradient according to our manual and preparing 8 working solutions for the standard. For accurate quantification, it's suggested to test both standards and samples in duplicate, enabling the testing of 40 samples per kit.
Reference
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---|---|---|---|---|
L00350
|
ToxinSensor™ Chromogenic LAL Endotoxin Assay Kit |
|
$199.00 |
|
L00351
|
ToxinSensor™ Gel Clot Endotoxin Assay Kit |
|
$147.00 1 Kit |
|
L00886
|
Benz-Neburase™ Nuclease ELISA Kit |
|
$498.00 96 Test |
|
L00942
|
AAV2 Titer Capsid ELISA Kit |
|
$498.00 96 Tests |
|
L00976
|
BSA ELISA Kit, 2G |
|
$498.00 96 Tests |
|
Z03695
|
Benz-Neburase™, tag-free |
|
$49.00 |
|
Z03708
|
Benz-Neburase™ GMP, tag free |
|
$75.00 |
|