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Description
This kit is intended for determining the presence of Trypsin-analog in biological product preparation processes, capable of identifying residual enzymes such as TrypLE™ from Thermo Fisher Scientific, Recombinant Trypsin from Sartorius, and HyrTryp™ from Cytiva.
Trypsin-analog are a non-animal origin high-purity recombinant fungal enzyme that is obtained from fermentation production of Fusarium oxysporum DSM2672. Trypsin-analog cleaves peptides on the C-terminal side of lysine and arginine residues as standard trypsin solutions do, which can be used to dissociate a variety of adherent cells, including but not limited to HEK293, A549, Normal Human Epidermal Keratinocytes (NHEKs) and Embryonic Stem Cell (ESC).
This kit is based on the solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect residual Trypsin-analog in the samples. A sheep polyclonal antibody specific to Trypsin-analog was employed in the assay to capture any remaining Trypsin-analog in the samples. Both the Calibration Standard (or test sample) and the HRP (Horseradish Peroxidase) labeled anti-Trypsin-analog 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 reaction, HRP catalyzed the oxidation of TMB by H2O2 to produce a blue 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 Trypsin-analog concentration in the Calibration Standard and the samples. The concentration of Trypsin-analog in the samples can be calculated using the dose-response curve.
| Components | Shipping Condition | Unit Size | Detection Method |
| Trypsin-analog Calibration Standard | 2-8℃ | 96 Tests | Sandwich ELISA |
| Anti-Trypsin-analog Microtiter Strips | |||
| Diluent | |||
| Wash Buffer Concentrate (10×) | |||
| Anti-Trypsin-analog:HRP (100×) | |||
| TMB Substrate | |||
| Stop Solution | |||
| Sealing Film |
Specifications
| Linearity & Range | 0.16-10.24 ng/mL, R2>0.990 |
| Accuracy | Recovery Rate= 94.0%-99.1% |
| LLOQ | 0.16 ng/mL |
| ULOQ | 10.24 ng/mL |
| Detection limit | 0.0319 ng/mL |
| Repeatability | 3.6%-4.4% (CV≤20%) |
| Specificity | No cross-reactivity with commonly protein analogs (e.g., recombinant human trypsin, recombinant bovine trypsin and recombinant porcine trypsin) and common engineering cell lines (e.g., MDCK, CHO, Vero, HEK293, E.coli, P.pastoris and Sf9 cells) |
| Robustness | Incubate at 25℃±5℃, CV≤ 20% |
| Instrument suitability not limited to Thermo Multiskan FC, Bio-Tek Synergy2 |
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