rChymoSelect™ MS Grade Protease Kit
Recombinant Chymotrypsin for Peptide Mapping and Proteomics
- High specificity for Tyr, Phe and Leu
- Eliminates non-specific cleavages
- Free from trypsin contamination
- Resistant to autoproteolysis for cleaner peptide maps
- Consistent lot-to-lot cleavage performance
- Does not require calcium for stability
- Engineered and optimized for peptide-mapping workflows
Catalog Number:
Size
Catalog Number: CS333204
Catalog Number: CS333205
Chymotrypsin Optimized for Mass Spec: Cleavage Specificity, Stability, Reproducibility
Chymotrypsin has long served as a critical orthogonal protease to trypsin in peptide mapping and proteomics. However, traditional chymotrypsins purified from bovine pancreas have the following shortcomings: high non-specific cleavages, susceptibility to autoproteolysis, calcium dependency and inconsistent lot-to-lot performance. These deficiencies complicate mass spec workflows, especially in regulated biotherapeutic characterization.
Our novel recombinant chymotrypsin, called rChymoSelect™ MS Grade Protease Kit, addresses all these pain points and sets a new benchmark for performance. This chymotrypsin belongs to a different class of chymotrypsin-like proteases than bovine chymotrypsin. Expressed in Pichia pastoris and purified to >98% homogeneity, it is engineered for high specificity, stability and reproducibility. It selectively cleaves at the C-terminus of Tyrosine, Phenylalanine and Leucine while avoiding non-specific cleavage at residues such as Alanine, Methionine, Histidine, Threonine, Valine, Serine, Glycine and others. rChymoSelect™ MS Grade Protease does not cleave after Tryptophan, enabling accurate analysis of tryptophan oxidation, a critical post-translational modification in biotherapeutic proteins.
Unlike conventional chymotrypsin, rChymoSelect™ MS Grade Protease Kit is resistant to autoproteolysis, independent of calcium for stability, and free of trypsin contamination. rChymoSelect™ MS Grade Protease Kit is not just a better chymotrypsin; it’s the first recombinant chymotrypsin designed specifically for mass spec-based peptide mapping and proteomics, delivering unmatched quality of peptide maps and consistency across every digest.
rChymoSelect™ MS Grade Protease Kit vs. Traditional Chymotrypsin
rChymoSelect™ MS Grade Protease Kit reduces non-specific cleavage, improving peptide analysis. Panel A. UV chromatograms comparing traditional chymotrypsin and rChymoSelect™ MS Grade Protease Kit digestion of a model substrate. Traditional chymotrypsin produces a complex peptide mixture with numerous semi- and non-specific cleavage products (blue arrows). In contrast, rChymoSelect™ MS Grade Protease Kit yields a simplified peptide profile with increased peak intensity for specific cleavages. Panel B. MS-based quantification of cleavage specificity shows that traditional chymotrypsin cleaves at multiple unintended sites. rChymoSelect™ MS Grade Protease Kit demonstrates high fidelity for phenylalanine (F), tyrosine (Y) and leucine (L) while avoiding cleavage at tryptophan.
rChymoSelect™ MS Grade Protease Kit exhibits resistance to autoproteolysis under standard digestion conditions. UPLC chromatograms show that traditional chymotrypsin generates numerous autoproteolytic fragments (blue arrows) following a 2-hour incubation at 25°C in 50mM Tris-HCl (pH 7.5). In contrast, rChymoSelect™ MS Grade Protease Kit remains intact under the same conditions, demonstrating enhanced resistance to autoproteolysis.
rChymoSelect™ MS Grade Protease Kit enables simpler analysis of W311 oxidation in panitumumab by avoiding cleavage after tryptophan. Mass spectrometry analysis of peptides containing W311 (tryptophan at position 311 in the heavy chain of panitumumab) following digestion with either traditional chymotrypsin or rChymoSelect™ MS Grade Protease Kit. Traditional chymotrypsin produces multiple overlapping peptides containing W311, complicating quantitation and retention time assignment. In contrast, rChymoSelect™ MS Grade Protease Kit generates cleaner fragmentation patterns, simplifying data interpretation and enhancing quantification accuracy for W311 oxidation.
rChymoSelect™ MS Grade Protease Kit extends sequence coverage in regions not covered by trypsin. Trypsin cleaves at lysine (K) and arginine (R), which sometimes can produce peptides that are either too short (when cleavage sites are clustered) or too long (in R/K-scarce, aromatic-rich regions), both of which may escape detection in MS analysis. For example, in the panitumumab heavy chain, trypsin digestion results in discontinuous sequence coverage across residues 317–347. In contrast, rChymoSelect™ MS Grade Protease Kit efficiently cleaves at phenylalanine (F), tyrosine (Y) and leucine (L), generating a well-sized, contiguous peptide in this region, complementing tryptic digestion and enhancing overall sequence coverage.
Key Differences Between Traditional Bovine Chymotrypsin and Recombinant rChymoSelect™ MS Grade Protease Kit.
|
Feature |
Traditional Chymotrypsin (Bovine) |
rChymoSelect™ MS Grade Protease Kit |
|
Source |
Bovine pancreas (animal-derived) |
Pichia pastoris (recombinant) |
|
Cleavage Specificity |
Tyr, Phe, Trp + non-specific sites |
Tyr, Phe, Leu, (Met*) |
|
Trp Cleavage |
Yes |
No |
|
Autoproteolysis |
High |
None** |
|
Calcium Dependency |
Yes |
No |
| Trypsin Contamination |
Possible |
None |
| Lot-to-Lot Reproducibility |
Moderate |
High |
| MS Compatibility |
Variable, lower peptide quality |
Optimized for clean peptide maps |
*Protein-dependent.
**Under recommended digestion conditions.
Protocols
No protocols available
Specifications
Catalog Number:
Contenido
| Item | Part # | Presentación |
|---|---|---|
rChymoSelect™ MS Grade Protease (Lyophilized) |
CS333201 | 1 × 25μg |
10X Digestion Buffer |
CS333203 | 1 × 1ml |
Limited Use Label License
BY USE OF THIS MATERIAL, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
Researcher may use this material for research use only; no transfer, or commercial use of this material is allowed. Commercial use means any and all uses of this material by a party in exchange for consideration, including, but not limited to
- (1) use in further material manufacture; and
- (2) resale of the material, whether or not such material is resold for use in research. Researcher shall have no right to modify or otherwise create variations or derivatives of the material. No other use of this material is authorized without the prior express written consent of Promega. Notwithstanding the foregoing, researcher may use this material in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the material. No other use of this material is authorized without the prior express written consent of Promega.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
Contenido
| Item | Part # | Presentación |
|---|---|---|
rChymoSelect™ MS Grade Protease (Lyophilized) |
CS333202 | 1 × 100μg |
10X Digestion Buffer |
CS333203 | 2 × 1ml |
Limited Use Label License
BY USE OF THIS MATERIAL, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
Researcher may use this material for research use only; no transfer, or commercial use of this material is allowed. Commercial use means any and all uses of this material by a party in exchange for consideration, including, but not limited to
- (1) use in further material manufacture; and
- (2) resale of the material, whether or not such material is resold for use in research. Researcher shall have no right to modify or otherwise create variations or derivatives of the material. No other use of this material is authorized without the prior express written consent of Promega. Notwithstanding the foregoing, researcher may use this material in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the material. No other use of this material is authorized without the prior express written consent of Promega.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
Resources
Featured Resource
Reinventing Chymotrypsin: A Novel Recombinant Analog for Accurate and Reliable Peptide Mapping of Biotherapeutic Proteins
rChymoSelect™ MS Grade Protease Kit demonstrates markedly improved cleavage specificity and resistance to autoproteolysis compared to its commercial counterparts. The novel, non-bovine chymotrypsin offers enhanced peptide mapping performance, which enables broader sequence coverage and more confident identification of post-translational modifications such as tryptophan oxidation.
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