Cation exchange–HPLC and mass spectrometry reveal C-terminal amidation of an IgG1 heavy chain

2007 ◽  
Vol 360 (1) ◽  
pp. 75-83 ◽  
Author(s):  
Keith A. Johnson ◽  
Kari Paisley-Flango ◽  
Bruce S. Tangarone ◽  
Thomas J. Porter ◽  
Jason C. Rouse
Keyword(s):  
Mass Spectrometry ◽  
Cation Exchange ◽  
Heavy Chain ◽  
Cation Exchange Hplc

scholarly journals Mass Spectrometry: A Tool for Enhanced Detection of Hemoglobin Variants

2008 ◽  
Vol 54 (1) ◽  
pp. 69-76 ◽  
Author(s):  
Peter Kleinert ◽  
Marlis Schmid ◽  
Karin Zurbriggen ◽  
Oliver Speer ◽  
Markus Schmugge ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cation Exchange ◽  
Single Point ◽  
Point Mutations ◽  
Reversed Phase ◽  
Chromatographic Methods ◽  
Globin Chains ◽  
Database Evaluation ◽  
Dna Variations ◽  
Cation Exchange Hplc

Abstract Background: More than 900 hemoglobin (Hb) variants are currently known. Common techniques used in Hb analysis are electrophoretic and chromatographic assays. In our laboratory, we routinely apply chromatographic methods. To ascertain whether Hb variants are missed with our procedures, we additionally analyzed all samples with mass spectrometry (MS). Methods: Database evaluation was performed using all entries made in the Hb variant database HbVar, and possible Hb variants were calculated based on DNA variations. During a 5-year period, we analyzed 2105 lysates with cation-exchange HPLC (PolyCAT A column) and reversed-phase HPLC and additionally with electrospray ionization or MALDI-TOF MS. Globin chains were identified by their molecular masses. Results: Database evaluation revealed that 43.2% of all possible Hbα- and β-chain variants were found to date (considering only single-point mutations). Currently, 68.2% of the possible charge difference variants and only 28.7% of the neutral variants are found. Among 2105 Hb samples we identified 4 samples with Hb variants that were detected only with the MS method; 2 were new Hb variants (Hb Zurich-Hottingen and Hb Zurich-Langstrasse). With cation-exchange HPLC, 1 sample was found to be a β-thalassemia and was identified by MS to be a β-variant (Hb Malay). More common variants, such as Hb C, Hb D, and Hb E, and thalassemias could not be detected with the MS method. Conclusions: Application of MS improves the sensitivity of Hb analysis. The combination of MS with electrophoretic and chromatographic methods is optimal for the detection of Hb variants.

scholarly journals The use of LC tandem mass spectrometry as part of a workflow for the screening and identification of hemoglobin variants. Characterization of Hb Ullevaal as an example

2017 ◽  
Vol 41 (3) ◽  
Author(s):  
Matthias Weber ◽  
Julia J.M. Eekels
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Cation Exchange ◽  
Peripheral Blood ◽  
Globin Gene ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
South Florida ◽  
Tandem Mass ◽  
Genetic Sequencing

AbstractBackground:About 2/3 of the hemoglobin (Hb) variants do not show a charge difference to the wildtype entity but most of them differ in hydrophobicity. In addition to cation exchange chromatography, globin differentiation by liquid chromatography-tandem mass spectrometry (MS) was introduced. Hb Ullevaal was chosen as one example to demonstrate the performance of the approach.Methods:Screening for Hb variants was performed using cation exchange HPLC. For globin separation reversed phase-LC/MS was performed. Tryptic digests of variants were separated on RP-HPLC with or without CID-fragmentation and database search for identification of mutation bearing fragments. Sequencing of the β-globin gene has been performed.Results:HbS, HbC, HbE, Hb South Florida and Hb Ullevaal show typical and distinct patterns in the globin LC/MS according to the theoretical protein data. The tryptic digest of Hb Ullevaal resulted in the identification of the respective mutated peptide βT9, which was confirmed by genetic sequencing.Conclusions:By the application of globin-LC/MS two more dimensions for the Hb identification are added, hydropathicity and protein mass. With this workflow as screening procedure for Hb variants it is expected to be able to detect and identify the majority of variants with the exception of highly unstable variants, which cannot be determined in the peripheral blood at all. A negative result makes the presence of a significant Hb variant in the peripheral blood improbable.

scholarly journals Signature Ions in MS/MS Spectra for Dansyl-Aminohexyl-QQIV Adducts on Lysine

Molecules ◽  
2020 ◽  
Vol 25 (11) ◽  
pp. 2659
Author(s):  
Lawrence M. Schopfer ◽  
Oksana Lockridge
Keyword(s):  
Mass Spectrometry ◽  
Heavy Chain ◽  
Plasma Proteins ◽  
Mass Spectrometry Data ◽  
Related Protein ◽  
Apolipoprotein A ◽  
Proteome Discoverer ◽  
Modified Peptides ◽  
Fluorescent Tags ◽  
Human Butyrylcholinesterase

Bacterial transglutaminase was used to label human plasma proteins with fluorescent tags. Protein lysines were modified with dansyl-epsilon-aminohexyl-Gln-Gln-Ile-Val-OH (dansylQQIV), while protein glutamines were modified with dansyl cadaverine. Labeled proteins included human butyrylcholinesterase, apolipoprotein A-1, haptoglobin, haptoglobin-related protein, immunoglobulin heavy chain, and hemopexin. Tryptic peptides were analyzed by LC-MS/MS on an Orbitrap Fusion Lumos mass spectrometer. Modified residues were identified in Protein Prospector and Proteome Discoverer searches of mass spectrometry data. The MS/MS fragmentation spectra from dansylQQIV-modified peptides gave intense peaks at 475.2015, 364.1691, 347.1426, 234.0585, and 170.0965 m/z. These signature ions are useful markers for identifying modified peptides. Human butyrylcholinesterase retained full activity following modification by dansylQQIV or dansyl cadaverine.

Inductively coupled plasma mass spectrometry coupled to cation exchange chromatography for the determination of trace nickel in alkaline electrolyte

2020 ◽  
Vol 35 (7) ◽  
pp. 1295-1299
Author(s):  
Lukas Miner ◽  
Diane Beauchemin
Keyword(s):  
Mass Spectrometry ◽  
Cation Exchange ◽  
Inductively Coupled Plasma ◽  
Exchange Chromatography ◽  
Alkaline Solutions ◽  
Alkaline Electrolyte ◽  
Cation Exchange Chromatography ◽  
Inductively Coupled ◽  
Plasma Mass Spectrometry

Cone corrosion by alkaline solutions with flow injection is prevented using cation-exchange chromatography coupled to inductively coupled plasma mass spectrometry.

Determination of Plasma Metformin by a New Cation-Exchange HPLC Technique

1999 ◽  
Vol 21 (3) ◽  
pp. 330 ◽  
Author(s):  
Anna Rita Bonfigli ◽  
Silvana Manfrini ◽  
Franco Gregorio ◽  
Roberto Testa ◽  
Ivano Testa ◽  
...  
Keyword(s):  
Cation Exchange ◽  
Hplc Technique ◽  
Cation Exchange Hplc

B6 vitamers: cation exchange HPLC

1990 ◽  
Vol 1 (12) ◽  
pp. 659-663 ◽  
Author(s):  
J.Dennis Mahuren ◽  
Stephen P. Coburn
Keyword(s):  
Cation Exchange ◽  
Cation Exchange Hplc

HbG-Honolulu interferes with some cation-exchange HPLC HbA1c assays

2016 ◽  
Vol 54 (3) ◽  
Author(s):  
Wan Ling Cheng ◽  
Siew Fong Neo ◽  
Suru Chew ◽  
Sunil Kumar Sethi ◽  
Tze Ping Loh
Keyword(s):  
Cation Exchange ◽  
Cation Exchange Hplc
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