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Product Description
SHENTEK Deoxyribonuclease I (DNase I) ELISA Kit is a 96-well, sandwich-format immunoassay for quantitative measurement of DNase I impurities in bioprocess samples and finished products (e.g., mRNA vaccines and therapeutics). The assay uses sheep polyclonal anti-DNase I capture antibody coated on microtiter strips, a biotinylated anti-DNase I detection antibody, and streptavidin-HRP for signal generation. TMB substrate is oxidized by HRP and measured at 450 nm (with 620–650 nm reference); concentration is determined using a 4-parameter logistic (4-PL) calibration curve. The kit is intended for Research Use Only (RUO) and supports testing from in-process samples through drug substance and drug product.
Technical Specifications
| Parameter | Details |
|---|---|
| Kit format | 96 tests/Kit |
| Assay principle | Double-antibody sandwich ELISA (capture antibody coated strips; biotinylated detection antibody; streptavidin-HRP; TMB substrate) |
| Linearity & Range | 0.5–32 ng/mL, R² ≥ 0.990 |
| Lower limit of quantitation (LLOQ) | 0.5 ng/mL |
| Calibration curve model | 4-parameter logistic (4-PL) |
| Typical 4-PL parameters (example) | A = 2.50521; B = -0.70719; C = 20.59525; D = -0.04360; R² = 0.99946 |
| Measurement wavelength | 450 nm (reference 620–650 nm) |
| Assay incubation times (per protocol) | Sample incubation: 1 hour (RT, 400–600 rpm); Biotinylated antibody: 45 min (RT); Streptavidin-HRP: 30 min (RT); TMB reaction: 15 min (RT); Post-stop incubation: 5 min |
| Specificity / Cross-reactivity | No cross-reactivity with T7 RNA Polymerase, RNase Inhibitor, BsaI; no cross-reactivity with host cell proteins from P. pastoris, MDCK, Vero, HEK293, Sf9, CHO, E. coli |
| Sample matrix / intended samples | In-process samples, harvested bulk, drug substance and drug product (e.g., mRNA production workflows) |
| Acceptable sample pH | pH 6.5–8.0 |
| Minimum recommended pipetting volume | 5 µL (recommended minimum for dilutions) |
| Storage | Store kit at 2–8 °C; protect light-sensitive components (TMB, streptavidin-HRP, biotinylated conjugate) |
Features
- High sensitivity and defined quantitation range: LLOQ 0.5 ng/mL with linear range 0.5–32 ng/mL and high curve fit (R² ≈ 0.999)
- High specificity: No cross-reactivity with key enzymes used in mRNA production (T7 RNA Polymerase, RNase inhibitor, BsaI) or common host cell proteins
- Ready-to-use coated strips: Microtiter strips pre-coated with affinity-purified sheep anti-DNase I capture antibody for consistent plate-to-plate performance
- Complete reagent set for ELISA: Includes calibration standard, diluent, wash buffer concentrate, biotinylated detection antibody, streptavidin-HRP, TMB substrate, and stop solution
- Designed for bioprocess monitoring: Suitable for quantitation of DNase I from in-process samples to final drug product, including mRNA workflows
Applications
- Quantitation of DNase I impurities during mRNA vaccine and therapeutic production
- Testing in-process samples, harvested bulk, drug substance and drug product
- Process development and release testing (research use)
- Method validation and spike-recovery studies for DNase I monitoring
Kit Contents
DNase I Calibration Standard (PNB016)1 × 350 µL; store at 2–8 °C; refer to tube label for details
Anti-DNase I Microtiter Strips (PNA016)8-well × 12 strips; strips pre-coated and packaged with desiccant; store in sealed bag at 2–8 °C (opened strips stable up to 90 days)
Diluent (PNE008)2 × 25 mL; store at 2–8 °C
Wash Buffer Concentrate (10×) (PNF001)2 × 25 mL; store at 2–8 °C; dilute 1:10 with ultrapure water before use
Anti-DNase I: Biotinylated Conjugate (100×) (PNG008)1 × 120 µL; sealed, protect from light; store at 2–8 °C; dilute 1:100 with Diluent fresh before use
Streptavidin-HRP (100×) (PNH002)1 × 140 µL; sealed, protect from light; store at 2–8 °C; dilute 1:100 with Diluent fresh before use
TMB Substrate (PND004)1 × 12 mL; sealed, protect from light; equilibrate to room temp 20 min before use; store at 2–8 °C
Stop Solution (PNI002)1 × 6 mL (1 M HCl); store at 2–8 °C; handle with goggles and protective equipment
Sealing Film (PNK001)3 pieces; use to cover strips during incubations; store at 2–8 °C
* Total: 96 tests/Kit | Method: ELISA
Attention
• For Research Use Only (not for clinical or diagnostic use).
• Store kit and components at 2–8 °C. Do not mix reagents from different lot numbers.
• Handle Stop Solution (1 M HCl) with care; avoid contact with eyes/skin; wear goggles and protective clothing.
• Protect light-sensitive reagents (TMB, streptavidin-HRP, biotinylated conjugate) from light.
• Avoid repeated freeze–thaw cycles of samples; for long-term storage keep samples at ≤ −70 °C.
• Samples should have pH between 6.5 and 8.0; values outside this range may cause abnormal results.
• Use fresh ultrapure water (temperature below 37 °C) for reagent preparation; if diluent or wash buffer has precipitate, heat to 37 °C until clear.
• Minimum recommended pipetting volume is 5 µL for dilution accuracy; avoid bubbles and foaming when pipetting.
• Centrifuge concentrated conjugates (100×) before use and prepare working solutions fresh; single-use recommended for some prepared solutions.
• Do not use samples containing sodium azide (NaN3) as it will inactivate HRP and underestimate DNase I levels.
• Complete plate reading within 30 minutes after stopping the reaction.
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What is the assay sensitivity and dynamic range?
The assay linear range is 0.5–32 ng/mL with an LLOQ of 0.5 ng/mL. Typical calibration fits a 4-parameter logistic model (example R² ≈ 0.999).
Q2: How should samples be prepared and stored prior to testing?
Samples must be clear/transparent; remove insoluble material by centrifugation or filtration. Avoid repeated freeze–thaw cycles; for long-term storage store at ≤ −70 °C. Adjust sample dilution to fit the calibration curve and ensure pH between 6.5–8.0.
Q3: Can this assay cross-react with enzymes used in mRNA production or common host cell proteins?
No. The kit has been tested and shows no cross-reactivity with T7 RNA Polymerase, RNase Inhibitor, BsaI, or host cell proteins from P. pastoris, MDCK, Vero, HEK293, Sf9, CHO, and E. coli.
Q4: What should I do if I observe high background signal?
Possible causes include reagent or equipment contamination, insufficient washing, or environmental contamination. Use freshly prepared reagents, increase wash volume/times, clean equipment with 75% ethanol, and avoid cross-contamination of wells.
Research Use Only
✔
Research Use Only (RUO) – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.
✔
Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
