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Product Description
This SHENTEK MDCK HCP ELISA Kit is a one-step, solid-phase, double-antibody sandwich ELISA for quantitative detection of residual host cell proteins (HCPs) from MDCK (Madin–Darby canine kidney) cells. A sheep polyclonal anti-MDCK HCP affinity-purified capture antibody is coated on microtiter strips and an HRP-conjugated sheep anti-MDCK HCP detection antibody (used simultaneously with standards/samples) enables colorimetric detection with TMB/H2O2 substrate. After incubation and washing, HRP catalyzes TMB oxidation (blue, max absorbance ~655 nm) and reaction is stopped to give a yellow product read at 450 nm (with 620–650 nm reference). Sample HCP concentrations are interpolated from a 4‑parameter logistic (4‑PL) calibration curve constructed from supplied calibration standards.
Designed for research use only (not for clinical use). Supplied as a 96-test kit with pre-coated microtiter strips, lyophilized calibration standard, HRP-conjugated detection antibody (100×), buffers and substrates. Typical application includes in-process testing and final product testing for MDCK-derived biologics including vaccines and influenza virus products.
Technical Specifications
| Parameter | Details |
|---|---|
| Kit format / throughput | 96 tests/Kit (8 wells × 12 strips) |
| Assay principle | One-step double-antibody sandwich ELISA, colorimetric detection (TMB, read at 450 nm with 620–650 nm reference) |
| Linearity & Range | 2–200 ng/mL (4‑PL curve), R2 ≥ 0.990 (typical R2 reported 0.9994) |
| Lower limit of quantitation (LLOQ) | 2 ng/mL |
| Calibration | Eight-point calibration curve (ST1–ST8) with 4‑parameter logistic (4‑PL) regression; example 4‑PL parameters: A=9.57058, B=-1.04748, C=399.32097, D=0.00304, R2=0.9994 |
| Typical OD data (reference) | Example OD450–620nm averages: 200 ng/mL = 3.237; 100 ng/mL = 1.931; 50 ng/mL = 1.089; 25 ng/mL = 0.610; 10 ng/mL = 0.312; 5 ng/mL = 0.210; 2 ng/mL = 0.148; 1 ng/mL = 0.133; 0 ng/mL = 0.111 |
| Specificity / Cross-reactivity | No cross-reactivity observed with E. coli, Pichia pastoris, or Sf9 cell HCPs |
| Sample types | In-process samples, harvested bulk, drug substance, drug product from MDCK-based processes |
| Recommended sample pH | pH 6.5–8.5 (outside this range may cause abnormal results) |
| Incubation conditions (typical) | Simultaneous addition of 1×Anti-MDCK:HRP and samples/standards; incubate on microplate thermoshaker at 600 rpm for 3 hours at room temperature; wash ×5; add TMB and incubate 15 minutes at RT; add stop solution and read |
| Readout | Absorbance at 450 nm with reference at 620–650 nm; OD450–reference used for curve fitting |
| Reagent stability (opened) | Anti‑MDCK HCP microtiter strips: store in bag with desiccant at 2–8 °C up to 60 days; reconstituted calibration standard: 2–8 °C up to 30 days |
Features
- One-step simultaneous detection: Detection antibody (HRP-conjugated) is added simultaneously with standards/samples to simplify workflow and reduce steps.
- High sensitivity: LLOQ 2 ng/mL with a linear dynamic range up to 200 ng/mL using 4‑PL fitting.
- MDCK-specific antibody coverage: Sheep polyclonal affinity-purified antibodies raised against MDCK HCPs provide broad coverage of host cell proteins from MDCK cells.
- Ready-to-use coated strips: Microtiter strips pre-coated with capture antibody in sealed bag with desiccant for ease of use and storage.
- Minimal cross-reactivity: Demonstrated no cross-reactivity with common non-MDCK host cells (E. coli, P. pastoris, Sf9).
Applications
- Quantification of residual host cell proteins (HCPs) in products manufactured with MDCK host cells
- Quality control testing of vaccines and influenza virus products produced in MDCK cells
- In-process monitoring (harvested bulk, intermediates)
- Testing of drug substance and drug product for MDCK-derived HCP contamination
Kit Contents
MDCK HCP Calibration Standard (lyophilized)Lyophilized; reconstitute with 500 μL Reconstitution Solution; reconstituted standard store 2–8 °C up to 30 days
Anti‑MDCK HCP Microtiter Strips (pre-coated)8 wells × 12 strips in sealed bag with desiccant; store at 2–8 °C; after opening store in bag with desiccant at 2–8 °C up to 60 days
Reconstitution Solution2 × 1.5 mL; room temperature equilibration before use
Diluent2 × 25 mL; store at 2–8 °C
Wash Buffer Concentrate (10×)2 × 25 mL; dilute 1:10 with ultrapure water before use; store at 2–8 °C
Anti‑MDCK:HRP (100×)1 × 120 μL HRP‑conjugated affinity‑purified sheep antibody; dilute 100× in Diluent before use; centrifuge before use; store at 2–8 °C
TMB Substrate1 × 12 mL; sealed, protect from light, equilibrate to RT before use; store at 2–8 °C
Stop Solution (1 M HCl)1 × 6 mL; corrosive—avoid contact with skin/eyes; store at 2–8 °C
Sealing Film3 pieces; room temperature storage
* Total: 96 tests/Kit | Method: ELISA (One-step double-antibody sandwich, HRP/TMB colorimetric)
Attention
• For Research Use Only (not for clinical diagnostic use).
• Equilibrate kit to room temperature for 20 minutes before use; return reagents to 2–8 °C after use.
• Use fresh 1×Wash Buffer and 1×Anti‑MDCK:HRP prepared immediately before use; prepare Anti‑MDCK:HRP by diluting 100× concentrate in Diluent and use immediately.
• Do not use Wash Buffer Concentrate or Diluent if cloudy or contains precipitates; heat to 37 °C to dissolve if necessary.
• Avoid repeated freeze-thaw cycles of samples; for long-term storage use ≤ -70 °C.
• Samples should be clear/transparent; remove particulates by centrifugation or filtration prior to assay.
• Maintain sample pH between 6.5 and 8.5 to avoid abnormal results.
• Do not mix reagents from different lot numbers.
• TMB Substrate should be colorless before use; discard if colored.
• Stop Solution is 1 M HCl — corrosive. Avoid contact with skin, eyes, and clothing.
• Avoid sodium azide (NaN3) in samples or buffers as it will inactivate HRP and lead to underestimated HCP levels.
• Read plate within 30 minutes after stopping reaction.
• Recommended minimum pipetting volume for accurate dilutions: 5 μL.
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: Is this kit suitable for clinical diagnostic use?
No. This kit is for Research Use Only (RUO) and is not intended for clinical diagnostic use.
Q2: How do I reconstitute the calibration standard?
Add 500 μL of the provided Reconstitution Solution to the lyophilized MDCK HCP Calibration Standard, gently invert 3–5 times, let stand 5 minutes until clear. Use the reconstituted standard to prepare the ST1–ST8 dilution series as described in the manual.
Q3: What is the assay sensitivity and dynamic range?
The assay LLOQ is 2 ng/mL and the validated dynamic range is 2–200 ng/mL using a 4‑parameter logistic (4‑PL) calibration curve.
Q4: What precautions should I take for samples and reagents?
Ensure samples are free of particulates (centrifuge/filtrate), maintain pH 6.5–8.5, avoid sodium azide, minimize freeze-thaw cycles, prepare 1× reagents fresh, and follow recommended incubation and washing steps carefully.
Research Use Only
✔
Research Use Only (RUO); not for clinical diagnostic use – 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.
