DT3C is a recombinant protein mainly used for in vitro evaluation of internalization efficiency of cancer cell towards monoclonal antibodies (mAb), thereby enabling the screening of more effective monoclonal antibodies. This protein consists of a diphtheria toxin (DT) protein without a receptor binding domain and Streptococcus protein G.
Application Areas of DT3C:
- In vitro evaluation of antibody internalization efficiency
- ADC drug research
- Antibody screening and evaluation
Technical Advantages of DT3C:
Compared to currently common techniques for evaluating antibody internalization efficiency, such as acid buffer elution, mAb-ZAP, pHrodo technology, the DT3C recombinant protein assay offers the advantages of simplicity in operation and precise quantification. It exhibits broad application prospects in the field of antibody internalization.
- Simple preparation: mAb-DT3C conjugates can be formed by incubating at room temperature for 30 minutes.
- Multi species applicability: It can can combine with any IgG from different species to form mAb-DT3C conjugates.
- Rapid evaluation: The internalization efficiency of mAb on cell surfaces can be quickly and accurately detected using techniques such as flow cytometry or fluorescence microscopy.
- High internalization efficiency: The internalization efficiency of antibodies is significantly higher than that of Mab ZAP.
In vitro detection of ADC antibody internalization efficiency
Reference: M. Yamaguchi et al. Biochemical and Biophysical Research Communications 454 (2014) 600-603.
DT3C Recombinant Protein from CUSABIO:
The DT3C recombinant protein from CUSABIO is expressed from Escherichia coli expression system. It's verified by SDS-PAGE and detected by SEC-HPLC, with a purity of over 90%.
After incubating different concentrations of DT3C protein with CCR8 antibody at 37°C for 30 minutes, the mixture was added to a culture plate and incubated for 48 hours. The range of EC50 values are 0.5795-1.400 μg/mL and 0.5608-1.318 μg/mL respectively.
How to use DT3C to detect antibody internalization efficiency?
- Cell culture conditions: Cultivate the cells to be tested under appropriate culture conditions.
- Antibody and DT3C conjugation: Incubate mAb and DT3C at room temperature for 30 minutes to form mAb-DT3C conjugates.
- Cell treatment: Add mAb-DT3C conjugates to the cell culture medium to interact with cells.
- Evaluation of internalization efficiency: Detect the internalization efficiency of mAb on the cell surface using flow cytometry, fluorescence microscopy, or other methods.