E.coli (Clonal strains) HCP ELISA Kit (One-step), BP-QP-E030-96
Escherichia coli (Clonal Strains) serves as the core prokaryotic host for biotherapeutic products including mRNA vaccines, viral vector-based gene therapies, and CAR-T products.
Product overview
Escherichia coli (Clonal Strains) serves as the core prokaryotic host for biotherapeutic products including mRNA vaccines, viral vector-based gene therapies, and CAR-T products. Host cell protein (HCP) residues generated during production and purification are critical process-related impurities. These trace contaminants may induce immunogenicity risks and compromise the purity and safety of finished products. HCP residual testing serves as a core assay item for process characterization, in-process control and product release. Critical raw materials including antigens and antibodies of this kit are independently manufactured by Zhuhai Biori Biotechnology Co., Ltd. Comprehensive method-performance validation of the kit has been conducted in accordance with the United States Pharmacopeia, Chinese Pharmacopoeia, European Pharmacopoeia and other regulations, satisfying the technical requirements for HCP testing stipulated by domestic and international biopharmaceutical regulatory authorities. The limit of detection (LOD) of the kit is approximately 0.249 ng/mL, and the limit of quantitation (LOQ) is approximately 0.861 ng/mL.
Features and applications
- Product type: One-step ELISA Kit
- Target Expression System: E.coli (DH5α, Top10, JM109, Stbl3)
- Assay Range: 1-243 ng/mL
- Time to result: <2.5h
- LOD: ~0.249 ng/mL
- LOQ: ~0.861 ng/mL
- Specificity: No cross-reactivity with Pichia pastoris, Sf9, MDCK and HEK293 HCP.
- The E.coli (Clonal Strains) HCP ELISA Kit (One-step) is intended for quantitative determination of host cell proteins (HCPs) in biotherapeutic products including mRNA vaccines, viral vector-based gene therapies, and CAR-T products.
- The kit can quantify residual HCP in purified intermediates, drug substance, and drug product, and can be used for process optimization, process monitoring, and product release.
- This is a generic kit and is not limited to a specific purification process. It broadly recognizes and detects HCPs originating from cloning strains DH5α, Top10, JM109, and Stbl3. For HCPs originating from other cloning strains, sample suitability should be verified.
- The kit is for research use only and not intended for clinical use.
Components
| Component | Quantity | Product No. |
|---|---|---|
| E.coli-C HCP Calibration Standards | 2 vials | BPA-002 |
| Anti-E.coli-C HCP Microplate Strips | 96 wells | BPB-002 |
| Anti-E.coli-C HCP:HRP (100×) | 120 μL × 1 tube | BPC-002 |
| TMB Substrate | 12 mL × 1 vial | BPD-001 |
| Stop Solution | 6 mL × 1 vial | BPE-001 |
| Diluent | 50 mL × 1 vial | BPF-001 |
| Wash Concentrate (20×) | 50 mL × 1 vial | BPG-001 |
| Plate Sealing Film | 2 pieces | BPI-001 |
| Instruction Manual | 1 copy | / |
Performance data
Calibration Curve (Assay Range)
Description: The standards supplied with the kit and one blank sample were tested. Each concentration was tested in triplicate, and three independent runs were performed. The Calibration curve was fitted using a four-parameter logistic model.
Conclusion: In three replicate experiments, the calibration curves fitted by the 4PL model showed R² ≥ 0.99, and both RE and CV were ≤ 15% at all concentration points (≤ 20% for the highest and lowest concentrations). The assay range of the kit is 1-243 ng/mL.
Accuracy
Description: A calibration curve was prepared over the validated range. Five quality control (QC) levels—ULOQ, high (H), medium (M), low (L), and LLOQ—were prepared, and each level was tested in triplicate.
Conclusion: The CV and RE of the measured concentrations were ≤20%.
Repeatability
Description: A calibration curve was prepared over the validated range. High, medium, and low QC samples were prepared and tested in eight parallel replicates.
Table 1 Repeatability Test Results
| Theoretical Concentrate | HQC-200ng/mL | MQC-100ng/mL | LQC-3ng/mL |
|---|---|---|---|
| No. of Tests | 8 | 8 | 8 |
| Mean Measured Concentration(ng/mL) | 189.4 | 95.1 | 3.4 |
| Concentration CV(%) | 3% | 3% | 5% |
Conclusion: The CV of the measured concentrations were ≤20%.
Inter-Analyst, Inter-Day, Inter-Instrument Precision
Description: Prepare calibration curve together with HQC, MQC, LQC samples for testing of inter-analyst, inter-day and inter-instrument precision.
Conclusion: The inter-analyst, inter-day and inter-instrument CV values were all below 20%, meeting the acceptance criteria.
Specificity
Description: A calibration curve was prepared over the validated range, and potential interferents were prepared in parallel. The low (L) QC level was selected as the final spike concentration.
Table 2 Specificity Test Results
| Sample | Sample concentration | Measured concentration(ng/mL) | Spike recovery |
|---|---|---|---|
| Pichia pastoris HCP | 1μg/mL | <LLOQ | 88% |
| HEK 293 HCP | 1μg/mL | <LLOQ | 83% |
| Sf9 HCP | 1μg/mL | <LLOQ | 82% |
| MDCK HCP | 1μg/mL | <LLOQ | 90% |
Conclusion: The kit showed no cross-reactivity with Pichia pastoris, Sf9, MDCK, HEK 293 host cell proteins at 1 µg/mL and the spike recoveries were within the acceptable range.
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | Store at 2-8°C. |
| Shipping | Refer to product label and shipping documentation. |
Downloads
Need this product lyophilized, in bulk, or in a custom format?
The feasibility assessment is free of charge. Send the assay brief and target format.
