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Related Concept Videos

NMR Spectroscopy Of Amines01:19

NMR Spectroscopy Of Amines

11.2K
In proton NMR spectroscopy, primary amines and secondary amines showcase their N–H protons as a broad signal in the chemical shift range between δ 0.5 and 5 ppm. The exact position in this range depends on several factors, including sample concentration, hydrogen bonding, and the type of solvent used. Since amine protons undergo fast proton exchange in solution, the protons are labile and therefore do not participate in any splitting with adjacent protons. Thus, the observed peak is...
11.2K
NMR Spectroscopy of Aromatic Compounds01:14

NMR Spectroscopy of Aromatic Compounds

6.4K
Aromatic compounds can be identified or analyzed using proton NMR and carbon‐13 NMR. Typically, aromatic hydrogens or hydrogens directly bonded to the aromatic rings are strongly deshielded by the aromatic ring current. Therefore, they absorb in the range of 6.5–8.0 ppm in proton NMR spectra. For instance, aromatic hydrogens directly bonded to the benzene ring absorb at 7.3 ppm. However, aromatic hydrogens of larger rings absorb farther upfield or downfield than the ideal range.
6.4K
NMR Spectroscopy of Benzene Derivatives01:34

NMR Spectroscopy of Benzene Derivatives

11.3K
Simple unsubstituted benzene has six aromatic protons, all chemically equivalent. Therefore, benzene exhibits only a singlet peak at δ 7.3 ppm in the 1H NMR spectrum. The observed shift is far downfield because the aromatic ring current strongly deshields the protons. Any substitution on the benzene ring makes the aromatic protons nonequivalent, and the protons split each other. The peak is, therefore, no longer a singlet and the splitting pattern and their associated coupling...
11.3K
NMR Spectroscopy: Chemical Shift Overview01:15

NMR Spectroscopy: Chemical Shift Overview

3.3K
The position of the absorption signal of a sample is reported relative to the position of the signal of tetramethylsilane (TMS), which is added as an internal reference while recording spectra. The difference between the absorption frequencies of the sample and TMS (in Hz) is divided by the spectrometer operating frequency (in MHz) to obtain a dimensionless quantity called the chemical shift. It is reported on the δ (delta) scale and expressed in parts per million.
For instance, the proton...
3.3K
NMR Spectroscopy: Spin–Spin Coupling01:08

NMR Spectroscopy: Spin–Spin Coupling

3.3K
The spin state of an NMR-active nucleus can have a slight effect on its immediate electronic environment. This effect propagates through the intervening bonds and affects the electronic environments of NMR-active nuclei up to three bonds away; occasionally, even farther. This phenomenon is called spin–spin coupling or J-coupling. Coupling interactions are mutual and result in small changes in the absorption frequencies of both nuclei involved. While nuclei of the same element are involved...
3.3K
NMR and Mass Spectroscopy of Carboxylic Acids01:30

NMR and Mass Spectroscopy of Carboxylic Acids

5.3K
In ¹H NMR spectroscopy, acidic protons (–COOH) of carboxylic acids are highly deshielded and absorb far downfield, at around 9–12 ppm. The chemical shift value depends on the concentration and solvent used.
While α protons of carboxylic acids absorb at 2–2.5 ppm, β protons absorb further upfield.
Carboxylic acids are easily identified by dissolving them in deuterium oxide, which results in a rapid exchange of the acidic protons with deuterium. This leads to the...
5.3K

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Overlapping Peptide Library to Map Qa-1 Epitopes in a Protein
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Mapping Antibody Epitopes by Solution NMR Spectroscopy: Practical Considerations.

Luca Simonelli1, Mattia Pedotti1, Marco Bardelli1

  • 1Institute for Research in Biomedicine, Universita' della Svizzera italiana (USI), Bellinzona, Switzerland.

Methods in Molecular Biology (Clifton, N.J.)
|May 2, 2018
PubMed
Summary
This summary is machine-generated.

Nuclear Magnetic Resonance (NMR) spectroscopy enables detailed identification of protein epitopes, which are crucial for antibody interactions and vaccine development. This method provides residue-level insights into antibody-antigen interfaces, aiding research and drug design.

Keywords:
AntibodyAntigenChemical shift mappingChemical shift perturbationEpitope mappingSolution NMR

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Immunology

Background:

  • Epitope identification is vital for basic research, pharmaceutical development, and vaccine design.
  • Antibody-antigen interactions form critical interfaces for biological recognition.
  • Understanding these interactions at a molecular level is key to advancing these fields.

Purpose of the Study:

  • To describe the application of Nuclear Magnetic Resonance (NMR) spectroscopy for the residue-level characterization of protein epitopes.
  • To provide insights into experimental protocols and practical considerations for using NMR in epitope mapping.
  • To highlight the advantages and limitations of NMR-based epitope identification.

Main Methods:

  • Utilizing Solution Nuclear Magnetic Resonance (NMR) spectroscopy.
  • Characterizing intermolecular interfaces at the residue level.
  • Focusing on antibody-antigen interactions for epitope mapping.

Main Results:

  • Demonstrated the capability of NMR spectroscopy for detailed epitope identification.
  • Provided a practical guide to experimental protocols for NMR-based epitope mapping.
  • Outlined the benefits and challenges associated with this NMR approach.

Conclusions:

  • NMR spectroscopy is a powerful tool for residue-level characterization of protein epitopes.
  • The described methods facilitate precise mapping of antibody-antigen binding sites.
  • This technique supports advancements in immunology, drug discovery, and vaccine development.