Peptide Coverage/Peptide Spectrum Match (PSM) Analysis Service

Protein sample identification generally only uses trypsin for protein digestion, with peptide coverage identified around 60%. To obtain the complete sequence information of the target protein, MtoZ Biolabs will use 6 commonly used proteinases in protein identification (trypsin, chymotrypsin, Asp-N, Glu-C, Lys-C, and Lys-N) to digest and identify the target protein. By obtaining more fragmented peptide fragments and stitching them together, 100% of the protein sequence is determined. MtoZ Biolabs employs the existing mass spectrometry technology platform to provide peptide mapping based on LC-MS/MS, which can be used to confirm whether the recombinant protein is fully expressed, and to detect whether the recombinant protein expression process has broken. Mainly used for the confirmation of the primary structure of proteins, by using multiple proteolytic reactions and LC-MS techniques for peptide sequencing, combined with proprietary analysis software, it can analyze the mass spectrometry data of any non-specific proteolytic products. Thus achieving 100% coverage of the measured target protein sequence.

Bock, T. et al. Front. Biosci. 2012.

Analysis Workflow

1. First Use 6 Kinds of Proteinases (Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C, and Lys-N) to Digest and Identify the Target Protein

2. Then Analyze the Sample Using Nano LC-MS/MS or UV LC-MS/MS

3. Compare the Full Sequence of Obtained Peptide Coverage/Peptidogram with the Theoretical Sequence you Provide

4. Obtain the Comparison Results

Service Advantages

1. The Combination of Specific and Non-specific Proteases, which can Achieve 100% Coverage of Any Protein/Peptide Identification

2. Each Peptide's Confirmation Includes the Quality of the Sample and the MS/MS Fingerprint Spectrum of the Sample

Sample Submission Requirements

1. Gel Samples and Solution Samples can be Used for the Above Sequencing

2. The Higher the Purity of the Sample, the Better, we Recommend Sample with Over 80% Protein

3. The Amount of Target Protein Sample Requires 10-20 μg

4. Protein Gel Samples Can also be Used for Peptide Coverage Determination, but the Sample Requires Special Pretreatment

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including: