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Related Concept Videos

NMR Spectroscopy Of Amines01:19

NMR Spectroscopy Of Amines

11.1K
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...
11.1K
NMR Spectroscopy of Aromatic Compounds01:14

NMR Spectroscopy of Aromatic Compounds

6.3K
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.
6.3K
NMR Spectroscopy of Benzene Derivatives01:34

NMR Spectroscopy of Benzene Derivatives

11.2K
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...
11.2K
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

3.2K
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...
3.2K
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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Diffusion Profiling of Therapeutic Proteins by Using Solution NMR Spectroscopy.

Bradley T Falk1, Yingkai Liang2, Mark A McCoy1

  • 1Mass Spectrometry and Biophysics, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ, 07033, USA.

Chembiochem : a European Journal of Chemical Biology
|December 6, 2018
PubMed
Summary

Nuclear Magnetic Resonance (NMR) diffusion methods create a "diffusion profile" to analyze therapeutic protein behavior in solution. This technique characterizes protein size and association states, aiding in drug development.

Keywords:
NMR spectroscopycharacterizationproteinssolution chemistrytranslational self-diffusion

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Area of Science:

  • Biophysics
  • Biochemistry
  • Pharmaceutical Science

Background:

  • Therapeutic protein development requires understanding structural and behavioral changes.
  • Nuclear Magnetic Resonance (NMR) spectroscopy is valuable for studying protein interactions and higher-order structures.
  • These higher-order structures can influence protein biological function and safety.

Purpose of the Study:

  • To introduce and validate a novel NMR diffusion method for characterizing therapeutic proteins.
  • To define a
  • diffusion profile
  • for interpreting protein behavior in solution.
  • To assess the utility of diffusion profiling as a complementary characterization technique.

Main Methods:

  • Utilized NMR diffusion techniques to analyze protein samples.
  • Developed the concept of a
  • diffusion profile
  • to represent protein solution behavior.
  • Compared diffusion profiling with traditional biophysical and NMR methods.

Main Results:

  • Diffusion profiles effectively capture complex diffusion behaviors in solution.
  • The method provides insights into the distribution of protein sizes and association states.
  • Diffusion profiling demonstrates a capacity to interpret intricate protein solution dynamics.

Conclusions:

  • Diffusion profiling is a valuable tool for characterizing therapeutic proteins in solution.
  • This method complements existing biophysical and NMR techniques.
  • Diffusion profiling enhances the understanding of protein behavior relevant to biological function and safety.