Integrated Protein Digestion HPLC
The benefits of using Perfinity iDP with the immobilized trypsin column are:
- Fast trypsin digestion in 1 to 4 minutes
- Continuous sample analysis on reusable trypsin column
- Reduced chymotrypsin activity and deamidation Insulin was digested within 1 minute using the trypsin column as shown in the figure at right.
Insulin was digested within 1 minute using the trypsin column as shown in the figure at right.* Optimal digestion time depends on proteins.
The optimized trypsin digestion buffer in combination with the trypsin column enables highly efficient digestion.
The figure to the right shows a comparison between the buffer. The optimized digestion buffer provided a higher peptide recovery rate with a short digestion time.
Two desalting columns allow parallel processing of samples. One desalting column is used to trap digested peptides, while peptides from the previously injected sample are eluted from the other desalting column.
This parallel sample processing allows analysis of up to 200 samples/day.
The figure to the right is an example of parallel sample processing, which increases throughput while maintaining reproducibility.
The Perfinity iDP software supports the total workflow from method development to analysis of samples
step 1 Method Settings
After selecting a method, users can quickly create and optimize methods by entering a few parameters, such as digestion time, column dimensions, and gradient time. Perfinity iDP methods are then automatically generated for running samples.
step 2 Batch file creation
Batch files for sample analysis arequickly generated. Users enter minimal information such as methods, data file names, and injection volumes.
step 3 Seamless connection with LabSolutions
Analysis results can be viewed in the LabSolutions post-run window, which can be accessed through the Perfinity iDP software.