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Product Description
This is a solid-phase, one-step double-antibody sandwich ELISA kit for quantitative detection of residual host cell proteins (HCPs) from Spodoptera frugiperda (Sf9) insect cells. A sheep polyclonal anti-Sf9 HCP antibody is coated on microtiter strips to capture HCPs; HRP-conjugated anti-Sf9 antibody is used as detection reagent. Calibration Standards and samples are added together with the HRP-conjugated antibody, followed by incubation, washing, TMB substrate development (HRP catalyzes oxidation of TMB producing color change) and stop solution addition. Absorbance is read at 450 nm with long-wavelength correction (620–650 nm). Concentrations are calculated using a 4-parameter logistic (4-PL) fit to the calibration curve. The kit is intended for research use only and is not for clinical diagnostic use.
Technical Specifications
| Parameter | Details |
|---|---|
| Package | 96 tests/Kit |
| Assay format | One-step double-antibody sandwich ELISA (capture antibody coated strips; simultaneous addition of HRP-conjugated detection antibody and samples/standards) |
| Linearity & Range | 3–243 ng/mL; 4-PL fit |
| LLOQ | 3 ng/mL |
| Calibration model | 4-parameter logistic (4-PL); example curve parameters provided (A, B, C, D) with R2≥0.990 |
| Typical calibration curve performance | Example R2 = 1.000 (reference data provided); reference standard concentrations: 0, 1, 3, 9, 27, 81, 243 ng/mL with corresponding OD values |
| Wavelength | Read at 450 nm with correction wavelength 620–650 nm |
| Incubation conditions | Sample + 1×Anti-Sf9:HRP incubation on thermoshaker: 3 hours at room temperature (600 rpm recommended); TMB substrate incubation: 30 minutes at room temperature |
| Specificity | No cross-reactivity reported with MDCK, Vero, HEK293T, CHO, E. coli and P. pastoris |
| Sample pH range | pH 6.5–8.5 (samples outside this range may cause abnormal results) |
| Wash buffer | Wash Buffer Concentrate (10×) supplied; dilute 1:10 with ultra-pure water to prepare 1×Wash Buffer; recommended 5 washes, 340 µL per well |
| Control & replicates | Recommend 2–3 replicates per sample and inclusion of NCS (negative control) and spiked recovery controls (SRC) for validation |
| Stability of opened components | Microtiter strips: store in bag with desiccant at 2–8 °C up to 30 days; reconstituted calibration standard: short-term 2–8 °C, long-term aliquot and store below -20 °C (avoid >3 freeze-thaw cycles) |
| Safety | Stop Solution is 1 M HCl (corrosive) — avoid contact with eyes, skin, clothing; avoid samples containing sodium azide (inactivates HRP) |
Features
- One-step format: Simultaneous addition of HRP-labeled detection antibody and standards/samples reduces hands-on steps and assay time.
- High sensitivity: Low limit of quantification (LLOQ) of 3 ng/mL with linear range up to 243 ng/mL.
- Broad antibody coverage: Sheep polyclonal anti-Sf9 HCP antibody provides broad recognition of Sf9 host cell proteins; coverage assessed by mainstream methods.
- Ready-to-use coated strips: Microtiter strips pre-coated with affinity-purified capture antibody in sealed bags with desiccant for convenience and reduced preparation time.
- Validated specificity: No cross-reactivity observed with common cell lines and microbial expression hosts tested (MDCK, Vero, HEK293T, CHO, E. coli, P. pastoris).
- Complete kit: Contains standards, detection antibody (HRP), diluent, wash buffer concentrate, substrate (TMB), stop solution and sealing film for a full ELISA workflow.
Applications
- Quantification of residual host cell proteins (HCPs) in products manufactured using Sf9 insect cells
- Testing of recombinant proteins produced in Sf9 cells
- Quality control of vaccines produced using baculovirus/Sf9 systems
- Residual HCP measurement for recombinant AAV vectors produced in BEVS
- In-process samples, harvested bulk, drug substance and drug product testing during R&D
Kit Contents
Sf9 HCP Calibration Standard (PNB011)Lyophilized; reconstitute with Reconstitution Solution; short-term 2–8 °C, aliquot and store < -20 °C for long-term (avoid >3 freeze-thaw cycles)
Anti-Sf9 HCP Microtiter Strips (PNA012)8 wells × 12 strips; store sealed in bag with desiccant at 2–8 °C; use within 30 days after opening
Reconstitution Solution (PNC002)2 × 1.5 mL; store at 2–8 °C
Diluent (PNE004)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 ultra-pure water before use
Anti-Sf9:HRP (100×) (PNN006)1 × 120 µL; affinity-purified sheep antibody conjugated to HRP; centrifuge before use; dilute 100× in Diluent immediately before use; store at 2–8 °C
TMB Substrate (PND004)1 × 12 mL; sealed, keep away from light; equilibrate to RT before use
Stop Solution (PNI002)1 × 6 mL; 1 M HCl; corrosive — store at 2–8 °C and handle with appropriate PPE
Sealing Film (PNK001)3 pieces; store at 2–8 °C
* Total: 96 tests/Kit | Method: One-step sandwich ELISA (solid-phase; HRP/TMB detection)
Attention
• For Research Use Only (not for clinical diagnostic use).
• Store the kit at 2–8 °C. Check expiration dates on labels.
• Equilibrate kit components to room temperature (≈20 minutes) before use; return to 2–8 °C after use.
• Reconstituted Sf9 HCP Calibration Standard: short-term 2–8 °C; aliquot and store < -20 °C for long-term; avoid >3 freeze–thaw cycles.
• Prepare 1×Anti-Sf9:HRP freshly by diluting the 100× stock; use immediately.
• TMB Substrate must be colorless before use; discard if colored. Protect from light during substrate incubation.
• Stop Solution is 1 M HCl — corrosive; avoid contact with eyes, skin and clothing; use appropriate PPE.
• Avoid samples containing sodium azide (NaN3) as it will inactivate HRP and lead to underestimation of HCP levels.
• Do not mix reagents from different lot numbers.
• Use fresh sterile/ultra-pure water (temperature ≤37 °C) for buffer preparation.
• Avoid drying wells before substrate addition and avoid bubbles when pipetting as they affect OD readings.
• Validate the assay and sample dilution strategy before routine use; recommended for qualified technicians only.
• Use single-use consumables; wipe benches and equipment with 75% ethanol before and after use to reduce contamination.
Quality Management & Certifications
Quality System
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Download CertificateQuality Advantages
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Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What is the assay sensitivity and dynamic range?
The assay linear range is 3–243 ng/mL with an LLOQ of 3 ng/mL. Calibration is performed using a 4-parameter logistic (4-PL) curve.
Q2: How should the Sf9 HCP Calibration Standard be stored after reconstitution?
Reconstituted standard can be stored short-term at 2–8 °C. For long-term storage, aliquot and store below -20 °C and avoid more than three freeze–thaw cycles.
Q3: Can this kit be used for samples from other host cells (e.g., CHO, HEK293)?
This kit is developed specifically for Sf9 host cell proteins. The kit was reported to show no cross-reactivity with MDCK, Vero, HEK293T, CHO, E. coli and P. pastoris, so it is not intended for quantifying HCPs from other host cell lines.
Q4: What precautions should be taken with sample buffers or additives?
Avoid samples containing sodium azide as it inactivates HRP. Ensure sample pH is between 6.5 and 8.5. If samples are turbid or contain particulates, clarify by centrifugation or filtration and validate sample dilutions.
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.
