Creative Biolabs has extensive experience in epitope mapping by nuclear magnetic resonance (NMR) analysis with the high degree of sophistication and technical expertise. Technicians in Creative Biolabs are always here to support clients’ projects and try to find the best solution for each project.
The antibody is the protein capable of recognizing antigen with high specificity and affinity. The antigen is recognized by atomic interactions between up to six highly variable loops called epitopes. Identification of epitopes is of great importance for the utilization of antibody, especially in the discovery and development of new therapeutics and vaccines design. A variety of methods for epitope mapping has been established up to now.
Nuclear magnetic resonance is particularly suited for determining the boundaries of epitopes recognized by antibodies. NMR analysis for identifying epitopes is based on the different mobility between the amino acid residues of the antigen that interact tightly with the antibody and peripheral residues that do not directly interact with the antibody. Epitopes interacting with the antibody undergoes significant changes in chemical shift. A detailed structural analysis for antigen-antibody complex is provided. Therefore, the nature of how the epitope is recognized could be accurately explained.
Basically, the NMR-based method for the epitope mapping is derived from the comparison between measurements of the spectrum. One is called HOmonuclear Hartmann-Hahn (HOHAHA), the other is called Rotating frame Overhause Effect Spectroscoty (ROSEY). The HOHAHA spectrum reveals the information about the dynamic states of the protons derived from the epitope where there is a chemical bond. ROESY, also known as CAMELOSIN (Cross-relaxation Appropriate for Minimolecules Emulatesd by Locked SPINs) is useful for determining which signals arise from protons that are close to each other in space. ROESY still could be used even if the protons are not literally bonded. Comparison of spectra between the tightly interacting and indirect residues enables the assignment of the mobile segments of the antigen.
Fig.1 The diagram of the workflow for epitope mapping based on NMR analysis.
NMR has greatly affected the way scientists are characterizing specificity and the affinity of a variety of molecules. Creative Biolabs provides a robust NMR platform regarding epitope mapping. We perform complete toolset comprising trusted solutions for every step we take. We now provide the optimized, validated platform in order to minimize and eliminate the trial-and-error phase. If you are meeting up with difficulties in determining epitopes, please feel free tocontact usby sending E-mail. A formal feedback will be sent back as soon as possible. We are always more than ready to reach out.