Protein Digestion for HPLC & MS Peptide Mapping

Fast, simple, reproducible protein digestion in about an hour

Use the latest advance in protein digestion for peptide mapping and quantitation applications. Thermo Scientific SMART Digest Kits make protein digestion:

  • Fast
  • Simple
  • Reproducible
  • Easy to automate

Protein digestion kits use optimized heat-stable enzymes on magnetic or non-magnetic beads. Immunoaffinity (IA) kits couple enzymatic digestion with IA capture on a single bead for high sensitivity. This innovation generates high confidence in your HPLC and LC/MS results, and it can be automated. SMART Digest Kits especially benefit biomarker and biopharmaceutical characterization and quantitation applications.

Fast, easy, reproducible

  • Simple 3-step protocol
  • Process samples in hours instead of days
  • Better reproducibility than in-solution digestion

Immunoaffinity (IA) capture

  • Enzymatic digestion with IA capture
  • Higher throughput
  • Greater sensitivity
     

Automate it
 


SMART Digest protein digestion kits for peptide mapping and peptide quantitation

SMART Digest kits: trypsin, chymotrypsin, or proteinase K protein digestion

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

PCR options

  Catalog Description Each Kit Includes
Order now 60109-101 SMART DIGEST KIT Trypsin 96 PCR tubes containing SMART Digest Resin
Collection Plate
Order now 60109-104 SMART DIGEST KIT Chymotrypsin 96 PCR tubes containing SMART Digest Resin
Collection Plate
Order now 60109-107 SMART DIGEST KIT Proteinase K 96 PCR tubes containing SMART Digest Resin
Collection Plate
Order now 60109-110 SMART DIGEST KIT Pepsin 96 PCR tubes containing SMART Digest Resin
Collection Plate
 
Order now 60109-102 SMART DIGEST KIT Trypsin 96 PCR tubes containing SMART Digest Resin
Collection Plate
96 Well Filter plate
Order now 60109-105 SMART DIGEST KIT Chymotrypsin 96 PCR tubes containing SMART Digest Resin
Collection Plate
96 Well Filter plate
Order now 60109-108 SMART DIGEST KIT Proteinase K 98 PCR tubes containing SMART Digest Resin
Collection Plate
96 Well Filter plate
Order now 60109-111 SMART DIGEST KIT Pepsin 99 PCR tubes containing SMART Digest Resin
Collection Plate
96 Well Filter plate
 
Order now 60109-103 SMART DIGEST KIT Trypsin 96 PCR tubes containing SMART Digest Resin
Collection Plate
SOLAµ 96 Well SPE microelution plate
Order now 60109-106 SMART DIGEST KIT Chymotrypsin 97 PCR tubes containing SMART Digest Resin
Collection Plate
SOLAµ 96 Well SPE microelution plate
Order now 60109-109 SMART DIGEST KIT Proteinase K 98 PCR tubes containing SMART Digest Resin
Collection Plate
SOLAµ 96 Well SPE microelution plate
Order now 60109-112 SMART DIGEST KIT Pepsin 99 PCR tubes containing SMART Digest Resin
Collection Plate
SOLAµ 96 Well SPE microelution plate

Magnetic bulk options

  Catalogue Description Each kit Includes
Order now 60109-101-MB SMART Digest Kit Trypsin
  • Bulk SMART Digest resin
  • SMART Digest buffer
Order now 60109-104-MB SMART Digest Kit Chymotrypsin
Order now 60109-107-MB SMART Digest Kit Proteinase K
       
Order now 60109-102-MB SMART Digest Kit Trypsin
  • Bulk SMART Digest resin
  • SMART Digest buffer
  • Collection plate
  • Filter plate
Order now 60109-105-MB SMART Digest Kit Chymotrypsin
Order now 60109-108-MB SMART Digest Kit Proteinase K
       
Order now 60109-103-MB SMART Digest Kit Trypsin
  • Bulk SMART Digest resin
  • SMART Digest buffer
  • Collection plate
  • SOLAμ SPE plate
Order now 60109-106-MB SMART Digest Kit Chymotrypsin
Order now 60109-109-MB SMART Digest Kit Proteinase K

Non-magnetic bulk options

  Catalogue Description Kits Include
Order now 60109-101-B SMART Digest Kit Trypsin
  • Bulk SMART Digest resin
  • SMART Digest buffer
Order now 60109-104-B SMART Digest Kit Chymotrypsin
Order now 60109-107-B SMART Digest Kit Proteinase K
       
Order now 60109-102-B SMART Digest Kit Trypsin
  • Bulk SMART Digest resin
  • SMART Digest buffer
  • Collection plate
  • Filter plate
Order now 60109-105-B SMART Digest Kit Chymotrypsin
Order now 60109-108-B SMART Digest Kit Proteinase K
       
Order now 60109-103-B SMART Digest Kit Trypsin
  • Bulk SMART Digest resin
  • SMART Digest buffer
  • Collection plate
  • SOLAμ SPE plate
Order now 60109-106-B SMART Digest Kit Chymotrypsin
Order now 60109-109-B SMART Digest Kit Proteinase K

Soluble enzyme option (for use with complex sample sets)

  Catalogue Description Kits Include
Order now 60113-101 SMART Digest Kit Soluble Trypsin
  • SMART Digest soluble trypsin
  • SMART Digest buffer

SMART Digest Bulk Fractionation Kits

  Catalogue Description
Order now 60114-101 SMART Digest Bulk Fractionation Kit, Protein A Non-Magnetic & Soluble Smart
Order now 60115-101 SMART Digest Bulk Fractionation Kit, Protein G Non-Magnetic & Soluble Smart
Order now 60114-102 SMART Digest Bulk Fractionation Kit, Protein A Non-Magnetic with SOLAµ/Collection plate & Soluble Smart
Order now 60115-102 SMART Digest Bulk Fractionation Kit, Protein G Non Magnetic with SOLAµ/Collection plate & Soluble Smart
Order now 60114-103 SMART Digest Bulk Fractionation Kit, Protein A Magnetic with SOLAµ/Collection plate  & SOLUBLE SMART
Order now 60115-103 SMART Digest Bulk Fractionation Kit, Protein G Magnetic with SOLAµ/Collection plate & Soluble smart
Order now 60114-104 SMART Digest Bulk Fractionation Kit, Protein A Magnetic & Soluble Smart
Order now 60115-104 SMART Digest Bulk Fractionation Kit, Protein G Magnetic & Soluble Smart
Order now 60116-101 SMART Digest Bulk Fractionation Kit, Protein A Magnetic standalone
Order now 60117-101 SMART Digest Bulk Fractionation Kit, Protein G Magnetic standalone
Order now 60118-101 SMART Digest Bulk Fractionation Kit Streptavidin Magnetic without Soluble Trypsin
Order now 60119-103 SMART Digest Bulk Fractionation Kit Streptavidin Magnetic with Soluble Trypsin, Collection plate and SOLAu
Order now 60119-104 SMART Digest Bulk Fractionation Kit Streptavidin Magnetic with Soluble Trypsin

SMART Digest Kits: low pH for MAM workflows

PCR options

  Catalogue Description Each Kit Includes
Order now 60109-101-LPH SMART DIGEST low pH KIT Trypsin 96 PCR tubes containing SMART Digest Resin
Collection Plate

Magnetic bulk options

  Catalogue Description Each Kit Includes
Order now 60109-101-MB-LPH SMART DIGEST low pH KIT Trypsin Magnetic beads 96 PCR tubes containing SMART Digest Resin
Collection Plate
Order now 60109-104-MB-LPH SMART DIGEST KIT CHYMOTRYPSIN Magnetic bulk low pH buffer 96 PCR tubes containing SMART Digest Resin
Collection Plate

Bulk options

  Catalogue Description Each Kit Includes
Order now 60109-101-MB-LPH SMART DIGEST low pH KIT Trypsin Magnetic beads 96 PCR tubes containing SMART Digest Resin
Collection Plate
Order now 60109-104-MB-LPH SMART DIGEST KIT CHYMOTRYPSIN Magnetic bulk low pH buffer 96 PCR tubes containing SMART Digest Resin
Collection Plate

SMART Digest IA Kits: trypsin digestion with immunoaffinity (IA) capture (streptavidin, protein A, protein G)

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.

Streptavidin (Av)

  Catalogue Description
Order now 60110-101 SMART Digest IA Kit, Streptavidin non-magnetic
Order now 60110-102 SMART Digest IA Kit, Streptavidin non-magnetic with Thermo Scientific SOLAµ SPE and collection plate
Order now 60110-103 SMART Digest IA Kit, Streptavidin magnetic with SOLAµ SPE and collection plate
Order now 60110-104 SMART Digest IA Kit, Streptavidin magnetic

Protein A

  Catalogue Description
Order now 60111-101 SMART Digest IA Kit, Protein A non-magnetic
Order now 60111-102 SMART Digest IA Kit, Protein A non-magnetic with SOLAµ SPE and collection plate
Order now 60111-103 SMART Digest IA Kit, Protein A magnetic with SOLAµ SPE and collection plate
Order now 60111-104 SMART Digest IA Kit, Protein A magnetic

Protein G

  Catalogue Description
Order now 60112-101 SMART Digest IA Kit, Protein G non-magnetic
Order now 60112-102 SMART Digest IA Kit, Protein G non-magnetic with SOLAµ SPE and collection plate
Order now 60112-103 SMART Digest IA Kit, Protein G magnetic with SOLAµ SPE and collection plate
Order now 60112-104 SMART Digest IA Kit, Protein G magnetic

Learn more about SMART Digest

Compare SMART Digest and in-solution techniques — SMART Digest saves time

In-solution protein digestion

SMART Digest vs. Conventional In-Solution Digest Protocol
Typical SMART Digest Digestion Times
Protein Digestion Time Protein Digestion Time
Insulin 4 min IgG in 50 µL plasma* 75 min
BSA < 5 min Ribonuclease A 150 min
Carbonic anhydrase < 5 min Thyroglobulin 240 min
Lysozyme < 5 min C-reactive protein 240 min
Apo-B 30 IgG 45 min      

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

Improve reproducibility

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.

Automate protein digestion for peptide mapping and quantitation

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.

Peptide mapping workflow

Peptide mapping workflow

Peptide quantitation workflow

Peptide quantitation workflow

SMART Digest webinars

SMART Digest webinar series

SMART Digest webinar series

Learn about four new technologies that dramatically improve peptide mapping workflows and increases confidence. 

  • Webinar 1: Don’t Miss a Thing on Your Peptide Mapping Journey
  • Webinar 2: Take the Smart Route to Protein Digestion
  • Webinar 3: Technologies for Improved Peptide Quantitation Workflow
  • Webinar 4: Peptide Separations with Pinpoint Precision

Attend Protein Digestion Webinar Series


SMART Digest videos

Watch this introductory animation to learn how SMART Digest kits provide easy, fast, and highly reproducible protein digestion.

 Watch SMART Digest video

Ajinomoto Althea is a contract development and manufacturing company. This is their experience using SMART Digest kits.

 Watch Anjimoto Althea testimonial

Save time and get better results with the SMART Digest Kit and automate your digestion with a KingFisher system with a touch of a button.

 Watch quick protocol video

SMART Digest Pepsin Kits tested and loved by Symphogen. Use the latest advance in protein digestion for peptide mapping and quantitation applications

 Watch video


Resources
CMD SchemaApp code