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Product Description
One-step double-antibody sandwich ELISA for quantification of residual CHO host cell proteins (HCPs) in CHO-expressed biologics. The kit uses affinity-purified sheep anti-CHO HCP capture antibody coated on microtiter strips and an HRP-conjugated polyclonal anti-CHO HCP detection antibody. Calibration standards and HRP-labeled detection antibody are added simultaneously to the coated wells, incubated, washed, then developed with TMB substrate (HRP catalyzes oxidation of TMB). After stopping the reaction with 1 M HCl, absorbance is read at 450 nm (with 620–650 nm reference). OD450 values correlate with HCP concentration and sample concentrations are calculated from a 4-parameter logistic (4PL) standard curve. No special sample pretreatment required; dilute samples into the calibration range as needed.
Technical Specifications
| Parameter | Details |
|---|---|
| Assay format | One-step double-antibody sandwich ELISA, 96-well format |
| Linearity & Range | 1–128 ng/mL, R² > 0.990 |
| Lower Limit of Quantification (LLOQ) | 1 ng/mL |
| Typical calibration model | 4-parameter logistic (4PL) fit (example: A=5.81069, B=-0.97488, C=200.99833, D=-0.00435, R²=0.99999) |
| Detection/Wavelength | Read at 450 nm (reference 620–650 nm) |
| Sample incubation time | 3 hours at room temperature (500–600 rpm recommended on thermoshaker) |
| Substrate incubation time | 15 minutes at room temperature (TMB) |
| Specificity | No cross-reactivity observed with MDCK, Vero, HEK293T, E. coli, P. pastoris and Sf9 cells |
| Sample types | Cell culture fluid, in-process samples, drug substance and drug product (clear/transparent samples; remove insolubles by centrifugation/filtration) |
| Calibration standards | 6-point calibration: 128, 64, 16, 4, 2, 1 ng/mL plus NCS (0 ng/mL) |
| Storage of kit | Store kit at 2–8 °C |
| Reconstituted standard stability | Reconstituted CHO HCP Calibration Standard: store at 2–8 °C up to 30 days or ≤ -20 °C with no more than 3 freeze-thaw cycles |
| Opened strip stability | Anti-CHO HCP microtiter strips: store sealed with desiccant at 2–8 °C for up to 60 days |
Features
- Sheep-derived polyclonal antibodies: Affinity-purified sheep anti-CHO HCP capture antibody and sheep polyclonal detection antibody for broad coverage of CHO HCPs.
- One-step assay workflow: Calibration standards and HRP-conjugated detection antibody are added simultaneously to coated wells to simplify setup.
- High sensitivity and broad dynamic range: LOQ 1 ng/mL with linear range up to 128 ng/mL and strong 4PL curve fit (R² > 0.99).
- Validated specificity: No cross-reactivity with several common host cell lines and expression systems (MDCK, Vero, HEK293T, E. coli, P. pastoris, Sf9).
- Ready-to-use plate strips: Pre-coated 8-well strips provided in a sealed bag with desiccant to reduce waste and allow flexible assay sizes.
- Standardized calibration: Six standardized calibration points and recommended 4PL analysis make quantitation straightforward.
Applications
- Quantitation of residual host cell proteins (HCPs) in CHO-expressed biologics (monoclonal antibodies, recombinant proteins, vaccines)
- In-process monitoring of upstream and downstream purification steps
- Release testing and process development research (research use only)
- Sample suitability testing and spiking/recovery studies for method validation
Kit Contents
Anti-CHO HCP Microtiter Strips (pre-coated)Store sealed with desiccant at 2–8 °C; after opening store strips in bag with desiccant at 2–8 °C up to 60 days
CHO HCP Calibration Standard (lyophilized) (PNB005) 2 bottlesReconstitute with 500 µL Reconstitution Solution; reconstituted standard: 2–8 °C up to 30 days or ≤ -20 °C (≤3 freeze-thaw cycles)
Reconstitution Solution (PNC002) 1×1.5 mL2–8 °C (use to reconstitute calibration standard)
Diluent (PNE004) 2×25 mL2–8 °C
Wash Buffer Concentrate (10×) (PNF001) 2×25 mL2–8 °C; dilute 1:10 with ultra-pure water before use; may crystallize at low temperature—dissolve at 37 °C if needed
Anti-CHO:HRP (100×) (PNN002) 1×120 µL2–8 °C; dilute 100× with Diluent immediately before use; centrifuge before use; discard after experiment (prepare fresh)
TMB Substrate (PND005) 1×12 mLStore at 2–8 °C; equilibrate to room temperature 20 min before use; keep away from light; discard if colored
Stop Solution (PNI002) 1×6 mL (1 M HCl)Store at 2–8 °C; handle with care (corrosive)
Sealing Film (PNK001) 3 piecesRoom temperature (for covering plate during incubation)
* Total: 96 tests/Kit | Method: ELISA (One-step double-antibody sandwich)
Attention
• For research use only (not for clinical diagnostic use).
• Use sterile disposable tips, tubes and reservoirs. Wipe bench and equipment with 75% ethanol as recommended.
• Do not mix reagents from different lot numbers.
• Reconstituted CHO HCP Calibration Standard and 1× Anti-CHO:HRP are unstable; prepare fresh and discard after use.
• Avoid samples containing sodium azide (NaN3) because it inactivates HRP and will underestimate HCP levels.
• Stop Solution is 1 M HCl—avoid direct contact with eyes, skin, and clothing; follow appropriate safety procedures.
• Recommend sample pH between 6.5 and 8.5; extreme pH may compromise measurements.
• Avoid repeated freeze–thaw cycles of samples; long-term sample storage recommended at -70 °C.
• Equilibrate TMB to room temperature for 20 minutes before use and protect from light during storage.
• Centrifuge Anti-CHO:HRP (100×) before dilution to avoid reagent loss; dilute 100× immediately prior to assay.
• Complete plate reading within 30 minutes after stopping the reaction.
• Avoid drying wells before substrate addition; seal plate to prevent evaporation during incubation.
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 sensitivity and dynamic range of this kit?
The kit LLOQ is 1 ng/mL with a linear quantification range of 1–128 ng/mL (4PL fit, R² > 0.99).
Q2: Can this kit detect HCPs from non-CHO host cells?
This kit is specifically developed for CHO-derived HCPs. It has been tested with no cross-reactivity to MDCK, Vero, HEK293T, E. coli, P. pastoris and Sf9, so it is not intended to quantify HCPs from non-CHO hosts.
Q3: How should samples and standards be stored?
Samples should be stored at ≤ -70 °C for long-term storage to avoid degradation and minimize freeze–thaw cycles. Reconstituted calibration standard: store at 2–8 °C up to 30 days or ≤ -20 °C with no more than 3 freeze–thaw cycles.
Q4: What are the key incubation times and temperatures for the assay?
Sample/standard incubation: 3 hours at room temperature on a thermoshaker (500–600 rpm). TMB substrate incubation: 15 minutes at room temperature. Read absorbance at 450 nm (reference 620–650 nm) after adding stop solution.
Research Use Only
✔
For 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.
