Related Experiment Video
Updated: Jun 17, 2026
10:50
Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System
Published on: May 1, 2019
Cyanogen bromide splitting of human immunoglobulin M
1Division of Clinical Immunology and Rheumatology, Department of Medicine, University of Alabama in Birmingham, University Station, Birmingham, Ala. 35294 USA.
Biochimica Et Biophysica Acta
|December 17, 2009
Summary
Researchers fragmented human Waldenstrom
Area of Science:
- Immunology
- Biochemistry
Background:
- Waldenstrom macroglobulinemia is a B-cell lymphoproliferative disorder characterized by the overproduction of IgM paraproteins.
- Understanding the structure of IgM is crucial for developing targeted therapies.
Purpose of the Study:
- To elucidate the primary structure of human Waldenstrom IgM (kappa) (Dau).
- To map the cyanogen bromide (CNBr) fragments of IgM, its mu chain, and its F(c)5mu and Fab micro fragments.
Main Methods:
- Cyanogen bromide (CNBr) cleavage of IgM and its fragments.
- Fractionation using gel filtration, ion exchange chromatography, and paper electrophoresis.
- Characterization via polyacrylamide disc electrophoresis, peptide mapping, amino acid composition, and primary structure determination.
Main Results:
- Two CNBr fragments were generated from the kappa chain, indicating one methionine residue.
- Five fragments were isolated from the S-sulfonated mu chain.
- The total number of CNBr fragments was determined to be eight or nine.
- A tentative arrangement of fragment order and disulfide bond locations was proposed.
Conclusions:
- The study provides insights into the primary structure of human Waldenstrom IgM.
- The fragmentation and characterization of CNBr pieces offer a framework for understanding IgM molecular architecture and disulfide bond organization.
More Related Videos
13:14Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
10:33Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow
Published on: January 8, 2014
Related Concept Videos
01:13¹H NMR: Complex Splitting
A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied first.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied first.
01:22Mass Spectrometry: Alkyl Halide Fragmentation
Chlorine isotopes exist as 35Cl and 37Cl in a 3:1 ratio, while bromine isotopes exist as 79Br and 81Br in a 1:1 ratio. The mass spectrum of alkyl halides typically produces two distinct molecular ion peaks, the molecular ion peak, [M], and the molecular ion plus two, [M + 2] peak. The relative heights of these two peaks are proportional to the isotopic abundance ratios of the halide. For example, 2‐chloropropane and 1‐bromopropane display two peaks with relative peak heights in a 3:1 and 1:1...
01:37NMR Spectroscopy of Benzene Derivatives
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 constants depend...
01:21Chemical Ionization (CI) Mass Spectrometry
The molecular ion peak of a molecule in the mass spectrum provides vital information for molecular identification. However, conventional electron impact ionization can lead to the rapid dissociation of some molecular ions before they reach the detector. A milder ionization method is required to increase the lifetime of such ionized analyte molecules. Chemical ionization (CI) is a gas-phase protonation reaction useful for mass-analyzing analyte molecules that are easily protonated to yield the...
02:41Formation of Halohydrin from Alkenes
An alkene, such as propene, reacts with bromine in the presence of water to yield a halohydrin. Halohydrins contain a halogen and a hydroxyl group attached to adjacent carbons. When the halogen is bromine, it is called a bromohydrin, while a chlorohydrin has chlorine as the halogen.
02:46Halogenation of Alkenes
Halogenation is the addition of chlorine or bromine across the double bond in an alkene to yield a vicinal dihalide. The reaction occurs in the presence of inert and non-nucleophilic solvents, such as methylene chloride, chloroform, or carbon tetrachloride.
Consider the bromination of cyclopentene. Molecular bromine is polarized in the proximity of the π electrons of cyclopentene. An electrophilic bromine atom adds across the double bond, forming a cyclic bromonium ion intermediate.
Consider the bromination of cyclopentene. Molecular bromine is polarized in the proximity of the π electrons of cyclopentene. An electrophilic bromine atom adds across the double bond, forming a cyclic bromonium ion intermediate.