Peptide Mapping Service

Peptide mapping is a widely used approach for analyzing biopharmaceuticals, including antibody, protein and peptides. The common peptide mapping process only uses trypsin for protein digestion, and the average coverage rate of identified protein is about 60%. In order to obtain the full sequence information of the targeted protein, MtoZ Biolabs uses up to 6 kinds of proteases for protein digestion (Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C and Lys-N). After protein digestion, sequences of peptide are mapped for full-length sequence of biopharmaceuticals. MtoZ Biolabs has also developed LC-MS/MS based peptide mapping technology, ensuring accurate peptide mapping analysis. This technology can be used to analyze the sequence of biopharmaceuticals and confirm complete expression of recombinant protein. With well-established technology and professional expertise, we can guarantee 100% peptide coverage and accurate data analysis.

 

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

Figure 1. Peptide Mapping Analysis of Biopharmaceuticals

 

Analysis Workflow 

1. Proper enzymes are selected for enzyme digestion and identification of target proteins.

2. Peptide analysis through LC-MS/MS or UV LC-MS/MS.

3. The total sequence of peptide coverage/peptide are mapped and compared with the theoretical sequence.

4. Delivery of report.

 

Service Advantages

1. Combination of specific and nonspecific proteases, achieving coverage of 100% of any protein (polypeptide).

2. Confirmation of each peptide is based on the MS/MS fingerprint of the sample, and is more accurate than MS analysis.

3. Protein sample 100% sequence validation service.

 

Sample Submission Requirements 

1. Gel and solution samples are all acceptable for peptide mapping service, but special pretreatment is required for gel sample.

2. The required amount of protein sample: 10-20ug.

3. Protein sample with purity of >80% is suggested.

 

Case Study 

In the each and every experiment, we will try to improve identification efficiency and reduce the cost in accordance with the analytical principle. In this case, the customer sent us an in vitro expressed antibody for peptide mapping analysis and comparison with the theoretical sequence. After receiving the sample, we first selected 3 proteases for digestion and designed subsequent sequence analysis experiments based on the theoretical sequences provided by our customers. Part of the experimental data are shown below, which proved that the antibody obtained was consistent with the theoretical sequence.

 

 

In the final report, we will also provide detailed data on the coverage of the peptides as shown in the following graph.

 

Figure 2. Case Study-Peptide Mapping Results

 

Deliverables

1. Experiment Procedure

2. Parameters of Liquid Chromatography and Mass Spectrometer

3. MS Raw Data Files

4. Peptide Mapping Results

5. Bioinformatics Analysis