B Cell-based Mass Spectrometry Assay

Mass spectrometry is an efficient and accurate analytical technique for biomolecules, particularly for proteins and peptides study. With cutting-edge instruments and experienced experts, Creative Biolabs offers mass spectrometry assay services for cancer epitope identification and characterization.

Background

Mass spectrometry is a cutting-edge tool for protein component analysis. This technique can determine the mass of various molecules, measuring the mass-to-charge ratio of their ions, the relative molecular mass of a compound, and the relative abundance and mass of isotopes of an element that can be interpreted to amino acid sequences and post-translational modification information of the epitope peptide. With tremendous improvement in the ionization approach, various mass spectrometry-based assays are used for determining antigen epitopes interacting with antibodies.

Fig.1 Immuno-MALDI mass spectrometry workflow.¹

Mass Spectrometry-based Cancer Epitope Analysis Service at Creative Biolabs

Mass spectrometry technique can determine both the amino acid sequences and structure of the epitope that binds to an antibody in solution. Creative Biolabs provides integrated cancer epitope analysis services, including sample preparation, mass spectrometry, and data analysis.

Workflow

Methodology

Mass Spectrometry-based Read-out Methods Available

The above sensitive, accurate, and reliable methods all have been used successfully for epitope mapping, while MALDI, ESI, and ToF are the main ionization methods used for mass spectrometric analysis. Except for mass determination, we also offer MS/MS to determine partial amino acid sequences.

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry
Electrospray ionization (ESI) mass spectrometry
Time-of-flight (ToF) mass spectrometry
Quadrupole time-of-flight (Q-ToF) mass spectrometry
Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometry
Ion traps mass spectrometry
Triple quadrupole mass spectrometry

Fig.2 The instrument of LC/MS. (Bohnert, et al., 2012)

Chemical Modification Methods

Chemical modification assay combined with mass spectrometry technology is used to identify the antigen-antibody binding interface on antigen targets for epitope analysis. The chemical modification methods include but are not limited to below:

Hydrogen/deuterium exchange (HDX)
Glycine ethyl ester labeling
Fast photochemical oxidation of proteins (FPOP)
Chemical cross-linking

Our Deliveries

By comparing the amino acid sequence of the mass spectrometric peptide with and without antibodies, we can successfully determine the binding epitope of the antibody. We deliver the results, including:

Report of relative label uptake on the protein sequence and three-dimension structure;
Indicate binding location on a 3D structure model;
Determine the linear or conformational epitope.

Highlights

Highlights (Creative Biolabs Original)

Creative Biolabs' professional team can offer clients comprehensive cancer epitope analysis services. Additionally, we can help you plan the research program that best fits your purpose and advances your research. Please contact us and let us know what we can do for you.

References

Froehlich, Bjoern.; et al. Systematic Optimization of the iMALDI Workflow for the Robust and Straightforward Quantification of Signaling Proteins in Cancer Cells. Proteomics. Clinical applications. 2020,14: 2000034.
Bohnert, Simone.; et al. Metabolomics in postmortem cerebrospinal fluid diagnostics: a state-of-the-art method to interpret central nervous system-related pathological processes. International Journal of Legal Medicine. 2012, 135(1): 183-191.

For Research Use Only | Not For Clinical Use