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Product Description
Solid-phase, one-step double-antibody sandwich Enzyme‑Linked Immunosorbent Assay (ELISA) for quantitative determination of residual host cell proteins (HCPs) from HEK293-derived host cells. The assay uses an affinity‑purified polyclonal capture antibody coated on microtiter strips and an HRP‑conjugated anti‑HEK293 HCP polyclonal detection antibody added simultaneously with calibration standards or samples. HRP catalyzes TMB oxidation to produce a colorimetric signal (read at 450 nm with long‑wavelength correction 620–650 nm). Sample HCP concentrations are determined by fitting a 4‑parameter logistic (4‑PL) calibration curve and applying the appropriate dilution factor.
Technical Specifications
| Parameter | Details |
|---|---|
| Assay format | One-step double-antibody sandwich ELISA (simultaneous addition of sample/standard and HRP conjugate) |
| Intended use | Determination of residual host cell proteins (HCPs) in products expressed in HEK293-derived host cells (e.g., recombinant proteins, cell and gene therapy products). For Research Use Only (RUO). |
| Package size | 96 tests/Kit |
| Linearity & Range | 6–540 ng/mL, R² ≥ 0.990 |
| Lower limit of quantification (LLOQ) | 6 ng/mL |
| Calibration standards | Concentrations: 0, 6, 18, 54, 135, 270, 540 ng/mL (prepared from lyophilized calibration standard) |
| Typical calibration curve (4‑PL) | A = 5.71467; B = -1.13651; C = 615.49064; D = 0.01444; R² = 0.99993 |
| Specificity | No cross-reactivity observed with E. coli HCP, P. pastoris HCP, Sf9 HCP, CHO HCP and Vero HCP |
| Assay incubation conditions | Sample/standard + 1× Anti‑HEK293:HRP incubate on microplate thermoshaker at room temperature, 500–600 rpm for 3 hours; TMB incubation 15 min at room temperature |
| Wavelength | Read at 450 nm with correction wavelength 620–650 nm (OD450‑620 recommended) |
| Washing | Wash 5 times with 340 µL of 1× Wash Buffer per well |
| Stop solution | 1 M HCl (Stop Solution), read within 30 minutes after termination |
| Sample requirements | Samples should be clear and transparent; remove insoluble material by centrifugation or filtration. Recommended sample pH 6.5–8.5. Avoid sodium azide in samples. |
| Storage (opened components) | Microtiter strips: store in vacuumed bag with desiccant at 2–8 °C up to 60 days; Reconstituted calibration standard: store at -20 °C, limit to ≤3 freeze‑thaw cycles |
Features
- High sensitivity: LLOQ 6 ng/mL enables detection of low levels of residual HEK293 HCPs.
- HEK293-specific detection: Affinity‑purified polyclonal antibodies raised against HEK293 HCPs provide specificity with no cross-reactivity to common host cell lines tested (E. coli, P. pastoris, Sf9, CHO, Vero).
- One-step assay workflow: Simultaneous addition of calibration standard/sample and HRP-conjugated detection antibody simplifies setup and reduces hands-on time.
- Pre-coated strip format: Microtiter strips pre-coated with capture antibody reduce reagent preparation and ensure lot-to-lot consistency (store with desiccant).
- Robust quantitative analysis: Calibration by 4‑parameter logistic regression with demonstrated high correlation (R² ≥ 0.99).
- Designed for bioprocess QC: Applicable to in-process samples, drug substance and drug product testing for HEK293‑derived processes.
Applications
- Quantitation of residual host cell proteins (HCPs) in products expressed in HEK293-derived cell lines
- Quality control testing for recombinant protein production using HEK293 hosts
- Assessment of HCP clearance during downstream purification
- Testing of in-process samples, drug substance and drug product for HEK293-derived manufacturing
- Assay development and validation during bioprocess development (method validation required)
Kit Contents
HEK293 HCP Calibration Standard (PNB008) — lyophilizedReconstitute with 500 µL Reconstitution Solution; reconstituted standard store at -20 °C (≤3 freeze‑thaw cycles)
Anti‑HEK293 HCP Microtiter Strips (PNA027) — 8 wells × 12 stripsPre-coated strips in vacuum bag with desiccant; store at 2–8 °C, reseal after use (up to 60 days for opened strips)
Reconstitution Solution (PNC002) — 2 × 1.5 mLRoom temperature for reconstitution; store per kit instructions (2–8 °C)
Diluent (PNE015) — 2 × 25 mLStore at 2–8 °C
Wash Buffer Concentrate (10×) (PNF001) — 1 × 25 mLStore at 2–8 °C; dilute 1:10 with ultrapure water before use
Anti‑HEK293:HRP (100×) (PNN017) — 1 × 120 µLStore at 2–8 °C (follow label); dilute 100× with Diluent immediately before use; centrifuge briefly before use
TMB Substrate (PND004) — 1 × 12 mLKeep sealed, protected from light; equilibrate to room temperature 20 min before use
Stop Solution (PNI002) — 1 × 6 mL (1 M HCl)Store at 2–8 °C; avoid contact with skin/eyes/clothing
Sealing Film (PNK001) — 3 piecesStore at 2–8 °C
* Total: 96 tests/Kit | Method: ELISA (One-step double-antibody sandwich)
Attention
• For Research Use Only (not for clinical diagnostic use).
• Use by qualified technicians only; validate assay before routine sample testing.
• Use sterile disposable tips, tubes and reservoirs; avoid cross-contamination (wipe work surfaces with 75% ethanol).
• Do not mix reagents from different lot numbers.
• Prepare 1×Wash Buffer by diluting 10× concentrate with ultrapure water; if cloudy, heat to 37 °C until clear.
• Centrifuge Anti‑HEK293:HRP (100×) briefly before dilution to avoid loss of reagent.
• HEK293 HCP Calibration Standard and freshly prepared 1× Anti‑HEK293:HRP are recommended for single use due to stability concerns.
• Minimum recommended pipetting volume is 5 µL; avoid excessive foaming during dilution.
• Stop Solution is 1 M HCl — avoid direct contact with eyes, skin and clothing; follow appropriate safety procedures.
• TMB Substrate must be colorless before use; discard if colored.
• Avoid samples containing sodium azide (NaN3) because it inactivates HRP and will lead to underestimation of HCP levels.
• Do not allow wells to dry before substrate addition; read plate within 30 minutes after stopping the reaction.
• Store unused microtiter strips sealed with desiccant to prevent contamination.
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 (LLOQ) and working range?
The assay LLOQ is 6 ng/mL with a linear range of 6–540 ng/mL (R² ≥ 0.990).
Q2: How should I store the kit and opened components?
Store the kit at 2–8 °C. Opened microtiter strips should be kept in the vacuum bag with desiccant at 2–8 °C (up to 60 days). Reconstituted calibration standard should be stored at -20 °C and limited to ≤3 freeze‑thaw cycles.
Q3: Can this kit detect HCPs from non‑HEK293 host cells?
This assay is specific for HEK293 HCPs and shows no cross-reactivity with E. coli, P. pastoris, Sf9, CHO or Vero HCP preparations. It is not intended for quantitation of non‑HEK293 host cell proteins.
Q4: What sample conditions or components can interfere with the assay?
Samples should be clear and free of insoluble material (centrifuge or filter as needed). Recommended pH is 6.5–8.5. Avoid sodium azide in samples as it deactivates HRP and causes underestimation of HCP levels.
Research Use Only
✔
Research Use Only (RUO) — Not for clinical use. Users should perform method validation per applicable guidelines (e.g., USP <1132>, EP <2.6.34>, ICH Q2(R2)). – 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.
