Peptide Mapping is a term used to describe a variety of methods applied to establishing the identity of a protein, hydrolysate, or a collection of peptides (as in a multiple-peptide vaccine product). Peptide Mapping may also be useful in proteomics and biomarker research. Blue Stream applies multiple methodologies to our Peptide Mapping programs. A partial selection of our Peptide Mapping capabilities includes:
Peptide Mapping: Optimized enzymatic digest with optimized chromatographic separation
Product variability is determined by detecting a peptide peak shift as compared to a reference standard. Such a study may be qualified or ICH validated and used as a "Peptide Map Fingerprint" as a part of a broader cGMP Lot Release Program.
Peptide Mapping: LC/MS
LC/MS Peptide Mapping may be used in conjunction with the optimized chromatographic Peptide Map. However, it may also be used together with a standard enzymatic digest or chemical hydrolysis, and standard chromatographic separation prior to MS analysis. The LC/MS method is often used for the verification of known amino acid sequence as compared to a theoretical sequence. This method is also useful for determination of site of glycosylation, disulfide bridge mapping, methionine oxidation, etc.
Peptide Mapping: LC/MS/MS
LC/MS/MS Peptide Mapping may be used to yield detailed information on each enzymatic peptide; its amino acid sequence, post translational modifications (PTM), glycan identification and structure, etc.
Blue Stream's Agilent 6538 QTOF allows for peptide mapping at better resolution with higher throughput.
Other applications of Peptide Mapping methods include:
- Peptide-profile analysis of multiple-peptide vaccine products
- Establishment of comparative peptide profiles (RM release programs) for hydrolysates used in cell culture/manufacturing
- Biomarker analysis
- Protein identification in complex mixtures (proteomics support)
- Comparative analysis of Follow-on Biologics
For LC/MS/MS Peptide Mapping, Blue Stream has incorporated Agilent's HPLC-Chip Cube technology, which is a biocompatible, nano-flow chromatographic system. The HPLC-Chip integrates columns, connection capillaries, and nanospray emitter directly on the polymer chip surface eliminating peak dispersion and providing uncompromised chromatographic performance. This is an excellent tool for high sensitivity applications.
Blue Stream will develop, qualify, and ICH-validate a Peptide Mapping method to yield Sponsor-specified scope of information. Please contact Blue Stream for additional information.