The kit employs a solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect residual CHO K1 host cell proteins in the samples. Polyclonal antibody specific to CHO K1 HCPs was employed in the assay to capture any remaining HCPs in the samples. Both the Calibration standards (or test samples) and the HRP (Horseradish Peroxidase) labeled anti-CHO K1 HCPs antibody were simultaneously added to the microtiter plate, which coated with the affinity purified capture antibody and followed by incubation and washing. Then TMB (3,3',5,5' -tetramethylbenzidine) substrate was added into the reaction, HRP catalyzed the oxidation of TMB by H2O2 to produce a blue product (maximum absorption peak at 655 nm). 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 standards and the samples. The concentration of CHO K1 HCPs in the samples can be calculated using the dose-response curve.
| 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: Biotinylated Conjugate (100×) | |||
| Streptavidin-HRP (100×) | |||
| TMB Substrate | |||
| Stop Solution | |||
| Sealing Film |
This kit utilizes 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 samples. Polyclonal antibody specific to E.coli (Protein expression strains) HCPs was employed in the assay to capture any remaining HCPs in the samples. Both the Calibration Standard and test samples were simultaneously added to the microtiter plate coated with affinity purified capture antibody, and followed by incubation and washing. The biotinylated antibody was added to the microtiter plate to bind the HCPs, then reacted with streptavidin labeled with HRP (Horseradish Peroxidase). TMB (3,3',5,5' -tetramethylbenzidine) substrate was added into reaction, HRP catalyzed the oxidation of TMB by H2O2 to produce a blue colored product (maximum absorption peak at 655 nm). Then the stop solution was added to terminate the enzymatic reaction, resulting in a yellow colored 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 samples. The concentration of HCPs in the sample can be calculated using the dose-response curve.
| Linearity & Range | 1-243 ng/mL, R²>0.990 |
| Accuracy | Recovery Rate= 87.1%-108.3% |
| LLOQ | 1.5 ng/mL |
| LOD | 0.0113 ng/mL |
| Repeatability | 3.5%-4.9% (CV≤20%) |
| 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±3℃, CV≤ 20% |
| Instrument suitability not limited to Thermo Multiskan FC, Bio-Tek Synergy2 |
SHENTEK® Residual Host Cell DNA Sample Preparation Kit
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