Protein Digestion for HPLC & MS Peptide Mapping

Choose between optimized heat-stable trypsin, chymotrypsin, or proteinase K enzymatic digestion for sequence-specific cleavage of your protein. The enzyme, on magnetic or non-magnetic beads, is activated at elevated temperatures for accelerated digestion under protein denaturing conditions.

• Trypsin cleaves the C-terminal side of lysine and arginine amino acid residues
• Chymotrypsin cleaves the peptide bonds formed by aromatic residues such as tyrosine, phenylalanine, and tryptophan
• Pepsin preferentially cleaves hydrophobic, preferably aromatic, residues in P1 and P1' positions. It will also preferentially cleave at the carboxyl side of phenylalanine and leucine and at the carboxyl side of glutamic acid residues
• Proteinase K cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked alpha amino groups

• SMART Digest User Manual

PCR options

SMART Digest Bulk Fractionation Kits

SMART Digest Kits: low pH for MAM workflows

PCR options

Magnetic bulk options

Bulk options

SMART Digest IA Kits perform trypsin digestion with immunoaffinity capture in a single well. The immunoaffinity moiety (streptavidin, protein A, or protein G) is typically employed to increase sensitivity by purifying your low-level proteins from complex biological matrices.

• SMART Digest ImmunoAffinity (IA) Kit User Manual

Obtain high-quality analytical results from mRNA digestions using SMART Digest RNase Kits. A significant advance in sample preparation for oligonucleotide mapping and quantitation, these kits provide fast and simple mRNA digestion with high reproducibility, high sensitivity, and high levels of data quality in a format that is compatible with automation.

200μL protein solution (100 μg/mL) at 70 °C
* IgG in plasma (17.5mg/mL total protein) at 70 °C

• SMART Digest compared to classic in-solution digestion of rituximab for in-depth peptide mapping characterization

The chromatogram overlay below is from three separate SMART digestions from the same mAb, conducted by three individual operators. The 15 marked peptides in each sample were used for inter-user/inter-day RSD value calculations.

• Digestion of rituximab for in-depth peptide mapping characterization

Peptide mapping and quantitation are fundamental workflows used in the development of biotherapeutics to ensure structural integrity, drug efficacy and patient safety. Current sample preparation techniques are complex, laborious, prone to error, and not amenable to automation impacting reanalysis rates, transfer, and implementation of methods at different sites. 

SMART Digest protein digestion provides a simple process on magnetic bead technology which allows it to be automated on platforms like the  Thermo Scientific King Fisher Duo to enable easy, fast, and highly reproducible results.