One-step ELISA Kit to quantify and characterize host cell proteins (HCPs) in products manufactured by expression in E.coli originated from BL21, such as interleukin (IL), recombinant human interferon (rhIFN), recombinant human tumor necrosis factor (rhTNF), growth-promoting factor (EGF/FGF/PDGF), et.al.
| Components | Shipping Condition | Unit Size | Detection Method |
| E.coli HCP-E Calibration Standard | 2-8℃ | 96 Tests | Sandwich ELISA |
| Anti-E.coli HCP-E Microtiter Strips | |||
| Reconstitution Solution | |||
| Diluent | |||
| Wash Buffer Concentrate (10×) | |||
| Anti-E.coli HCP-E:HRP (100×) | |||
| TMB Substrate | |||
| Stop Solution | |||
| Sealing Film |
This kit employs a solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect residual E.coli (Protein expression strains) host cell proteins (HCPs) in the sample. Polyclonal antibody specific to E.coli (Protein expression strains) HCPs was employed in the assay to capture any remaining HCPs in the sample. Both the Calibration standards (or test sample) and the HRP (Horseradish Peroxidase) labeled with anti-E.coli (Protein expression strains) HCP antibody were simultaneously added to the pre-coated microtiter plate, and followed by incubation and washing steps. Then TMB (3,3',5,5'-tetramethylbenzidine) substrate added for reaction, HRP catalyzed the oxidation of TMB by H2O2 to produce a blue product (maximum absorption peak at 655 nm). Afterwards the stop solution is added to terminate the enzymatic reaction, resulting in a yellow color product (maximum absorption peak at 450 nm). The absorbance values at 450 nm wavelength were positively correlated with the HCPs concentration in the Calibration standard and the sample. The concentration of E.coli (Protein expression strains) in the sample can be calculated using the dose-response curve.
| Linearity & Range | 1.5-243 ng/mL, R²>0.990 |
| Accuracy | Recovery Rate= 83.8%-110.2% |
| LLOQ | 1.5 ng/mL |
| LOD | 1.19 ng/mL |
| Repeatability | 4.5%-8.6% (CV≤15%) |
| Antibody Coverage (IMBS-2D) | 70.6%-100% |
| Antibody Coverage (IMBS-LC/MS) | 88.8% |
| Specificity | No cross-reactivity with CHO, Vero, HEK293T, Hansenula polymorpha HCPs |
| Robustness | Incubate at 25±5℃, CV≤ 15% |
| Instrument suitability not limited to Thermo Multiskan FC, Bio-Tek Synergy2 |
SHENTEK® Residual Host Cell DNA Sample Preparation Kit
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