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Product Description
Porcine Trypsin ELISA Kit is a solid-phase, double-antibody sandwich Enzyme-Linked Immunosorbent Assay (ELISA) for quantitative detection of residual porcine trypsin in biological products and downstream purification samples. A sheep polyclonal anti-trypsin capture antibody is coated on microtiter strips. Samples and calibration standards are added to the coated wells, followed by a biotinylated anti-trypsin detection antibody and Streptavidin-HRP. HRP catalyzes TMB oxidation to produce a colorimetric signal (read at 450 nm with a 620–650 nm reference). The kit includes lyophilized calibration standards, pre-coated strips, reagents for detection (biotinylated antibody and Streptavidin-HRP), TMB substrate and stop solution. The assay is intended for research use only to detect residual recombinant or pancreas-derived porcine trypsin in harvested bulk, in-process samples, drug substance and drug product.
Technical Specifications
| Parameter | Details |
|---|---|
| Format | Sandwich ELISA (solid-phase) |
| Sample volume per well | 100 µL |
| Detection wavelength | 450 nm (reference 620–650 nm) |
| Linearity & Range | 0.040–2.56 ng/mL (R² ≥ 0.990) |
| Lower Limit of Quantification (LLOQ) | 0.04 ng/mL |
| Calibration standards (nominal concentrations) | ST1 2.56 ng/mL, ST2 1.28 ng/mL, ST3 0.64 ng/mL, ST4 0.32 ng/mL, ST5 0.16 ng/mL, ST6 0.08 ng/mL, ST7 0.04 ng/mL, NCS 0 ng/mL |
| Calibration model (example) | 4-parameter logistic (4-PL); example fit parameters provided (A=2.16282, B=-1.00528, C=1.54458, D=-0.01333, R²=0.99996) |
| Specificity / Cross-reactivity | No cross-reactivity observed with CHO, MDCK, Vero, HEK293, E. coli and Hansenula polymorpha |
| Plate washing | Use 1× Wash Buffer; typical wash volume 340 µL/well; wash 3 times between steps (5 times before TMB) |
| Incubation times (typical per protocol) | Sample incubation: 1 hour at RT (500 rpm); Anti-Trypsin:Biotin incubation: 45 min at RT; Streptavidin-HRP incubation: 30 min at RT; TMB incubation: 15 min at RT; post-stop incubation: 5 min |
| Plate layout / replicates | Recommend 2–3 replicates per sample and inclusion of spiked recovery controls |
| Storage of kit | Store kit at 2–8 °C. Check expiration date on labels. |
| Storage of opened components | Anti-Trypsin microtiter strips: 2–8 °C (in bag with desiccant) up to 90 days; Reconstituted Trypsin Calibration Standard: < -20 °C up to 90 days (≤3 freeze-thaw cycles) |
| pH recommendation for samples | Recommended sample pH 6.0–8.0 |
Features
- High sensitivity: LLOQ 0.04 ng/mL and linear range down to 0.04 ng/mL enables detection of low residual trypsin levels.
- High specificity: Sheep polyclonal anti-trypsin antibodies with no observed cross-reactivity to common host cell lines and microorganisms (CHO, MDCK, Vero, HEK293, E. coli, Hansenula).
- Complete kit format: Includes pre-coated strips, lyophilized standards, detection reagents (biotinylated antibody, Streptavidin-HRP), TMB substrate and stop solution for a 96-test kit.
- Quantitative readout: 4-parameter logistic calibration recommended for accurate quantitation; absorbance read at 450 nm with reference wavelength.
- Designed for downstream process monitoring: Suitable for testing residual recombinant porcine trypsin and pancreas-derived trypsin in in-process and final product samples.
Applications
- Determination of residual porcine trypsin in downstream purification of biologics
- Testing harvested bulk, in-process samples, drug substance and drug product for residual trypsin
- Process development and validation for recombinant protein production where trypsin is used (e.g., cell dissociation, activation steps)
- Research applications requiring quantitation of porcine trypsin contamination
Kit Contents
Trypsin Calibration Standard (lyophilized)PNB006, 2 bottles; store lyophilized per label; reconstituted standard < -20 °C (up to 90 days)
Anti-Trypsin Microtiter Strips (pre-coated)PNA006, 8 wells × 12 strips; store in vacuumed bag with desiccant at 2–8 °C; use within 90 days after opening
Reconstitution SolutionPNC003, 1 × 240 μL; room temperature handling; used to dissolve calibration standard
Wash Buffer Concentrate (10×)PNF001, 3 × 25 mL; dilute 10× with ultra-pure water to 1×; store 2–8 °C
Anti-Trypsin:Biotin (100×)PNG006, 1 × 140 μL; centrifuge before use; dilute 100× in 1× Wash Buffer; prepare fresh
Streptavidin-HRP (100×)PNH002, 1 × 140 μL; centrifuge before use; dilute 100× in 1× Wash Buffer; prepare fresh
TMB SubstratePND002, 1 × 12 mL; equilibrate to RT 20 min before use; TMB should be colorless
Stop Solution (1 M HCl)PNI002, 1 × 6 mL; corrosive — avoid contact with eyes, skin, clothing
Sealing FilmPNK001, 3 pieces; use to prevent evaporation during incubations
* Total: 96 tests/Kit | Method: ELISA (double-antibody sandwich)
Attention
• For research use only (not for clinical or diagnostic use).
• Store kit at 2–8 °C. Check expiration dates on labels.
• Reconstituted calibration standard: store below -20 °C up to 90 days; avoid more than 3 freeze-thaw cycles.
• Anti-Trypsin microtiter strips: store in sealed bag with desiccant at 2–8 °C; use within 90 days after opening.
• Prepare 1× Wash Buffer freshly by diluting 10× concentrate with ultra-pure water; if cloudy, warm at 37 °C until clear.
• Dilute Anti-Trypsin:Biotin and Streptavidin-HRP 100× in 1× Wash Buffer immediately before use; centrifuge concentrates before dilution.
• Use TMB substrate only if colorless; equilibrate to RT for 20 minutes before use.
• Stop Solution is 1 M HCl — corrosive. Avoid contact with skin, eyes and clothing; handle in appropriate PPE.
• Avoid samples containing sodium azide (NaN3) because it will deactivate HRP and cause underestimation of trypsin levels.
• Recommend sample pH between 6.0 and 8.0; outside this range may produce abnormal results.
• Avoid repeated freeze-thaw of samples; long-term storage recommended at -70 °C.
• Do not mix different kit lots in the same assay.
• Maintain accurate pipetting (minimum recommended volume 5 µL for dilutions) and avoid bubbles.
• Trypsin Calibration Standard Solutions, Biotinylated Antibody Solution and Streptavidin-HRP Solution are recommended for single use due to stability concerns.
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's linear range and sensitivity?
The assay linear range is 0.040–2.56 ng/mL with an LLOQ of 0.04 ng/mL (example R² ≥ 0.990).
Q2: How should I store opened strips and reconstituted standards?
Store opened anti-trypsin microtiter strips in the sealed bag with desiccant at 2–8 °C for up to 90 days. Reconstituted Trypsin Calibration Standard should be stored below -20 °C for up to 90 days and subjected to no more than three freeze-thaw cycles.
Q3: What if I observe high background or abnormal OD values?
Common causes include cross-contamination, insufficient washing, equipment contamination, or environmental contamination. Use freshly prepared reagents, increase wash volume or wash cycles, clean equipment with 75% ethanol, and ensure proper plate sealing and handling to reduce background.
Q4: Which sample types and pH are suitable?
Suitable for harvested bulk, in-process samples, drug substance and drug product. Recommended sample pH is between 6.0 and 8.0. Remove insoluble material by centrifugation or filtration and avoid repeated freeze-thaw cycles.
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.
