This kit is suitable for quantitative of porcine trypsin in downstream purification processes of biological products, capable of identifying commonly available recombinant porcine trypsin and trypsin derived from porcine pancreas.
Porcine trypsin, a serine protease, specifically cleaves peptide bonds at the C-terminal of lysine and arginine. It is primarily used in the production of recombinant insulin, activation of virus particles, and cell processing in vaccine manufacturing processes.
This kit is based on the solid-phase Enzyme-linked Immunosorbent Assay (ELISA) with a double-antibody sandwich technique to detect residual trypsin in the samples. A sheep polyclonal antibody specific to trypsin was employed in the assay to capture any remaining trypsin in the samples. Both Calibration standards and test samples were added to the microtiter plate pre-coated with capture antibody. After washing out unbound proteins, the Detection Antibody and biotin conjugated anti-Trypsin antibody were added into the well to form the complexe. Following incubation and washing, the horseradish peroxidase conjugated Streptavidin(Streptavidin-HRP) is added. After another incubation and washing step, 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 concentration in the calibration standards and the samples. The concentration of Trypsin in the samples can be calculated using a dose-response curve.
| Components | Shipping Condition | Unit Size | Detection Method |
| Trypsin Calibration Standard | 2-8℃ | 96 Tests | Sandwich ELISA |
| Anti-Trypsin Microtiter Strips | |||
| Reconstitution Solution | |||
| Wash Buffer Concentrate (10×) | |||
| Anti-Trypsin:Biotin (100×) | |||
| Streptavidin-HRP (100×) | |||
| TMB Substrate | |||
| Stop Solution | |||
| Sealing Film |
| Linearity & Range | 0.04-2.56 ng/mL, R2>0.990 |
| Accuracy | Recovery Rate= 85.7%-111.0% |
| LLOQ | 0.04 ng/mL |
| ULOQ | 2.56 ng/mL |
| Detection limit | 0.0144 ng/mL |
| Repeatability | 4.1%-7.7% (CV≤20%) |
| Specificity | No cross-reactivity with commonly protein analogs (e.g., Ovalbumin and BSA from two different manufacturers, respectively), common engineering cell lines and engineering bacteria protein (e.g., MDCK, CHO, Vero, 293T, E.coli, and P.pastoris) |
| Robustness | Incubate at 25℃±5℃, CV≤ 20% |
| Instrument suitability not limited to Thermo Multiskan FC, Bio-Tek Synergy2 |
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