HPLC-Based Quality Peptide Mapping Assays Service

HPLC-based quality peptide mapping assays integrates the separation capabilities of HPLC with the qualitative and quantitative capabilities of MS. Utilizing the LC-MS/MS platform, which combines HPLC with tandem high-resolution MS, peptides are effectively separated by HPLC and their mass and charge are measured by MS. This allows for the identification of peptide segments corresponding to each chromatographic peak, forming a characteristic peptide fingerprint map.HPLC- based quality peptide mapping assays provides comprehensive information about the sample.

 

MtoZ Biolabs has established an quality peptide mapping assays platform by combining a HPLC with a high-resolution mass spectrometer, and offers comprehensive HPLC peptide mapping analysis services.

 

Experimental Instruments

1. Liquid Chromatograph: UltiMate3000

2. High-Resolution Mass Spectrometer: AB SCIEX 5600+

 

Applications 

1. Assessing batch-to-batch stability and identifying differences between batches and standard samples using mirror images, with precise identification of inconsistent regions.

2. Qualitative analysis of protein variants, localization of disulfide bonds, analysis of glycosylation sites, and determination of protein modification sites.

 

Case Study

After the sample was separated by HPLC, the data were collected by mass spectrometer and the peptide mapping was obtained as follows:

 



Figure 1. Total Ion Flow Diagram of The Test Sample

 

The original data of MS were retrieved and analyzed by software, and the corresponding relationship between peptide information and chromatographic peak was identified as follows:

 



Figure 2. The Corresponding Table of Peptide Information and Chromatographic Peak of The Test Sample

 



Figure 3. Sequence Overlays

 

Deliverables

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. Detailed Information on HPLC-Based Quality Peptide Mapping Assays

4. Mass Spectrometry Images

5. Raw Data