Isoelectric focusing in immobilized pH gradients with carrier ampholytes added for high-resolution phenotyping of bovine β-lactoglobulins: Characterization of a new genetic variant

1988 ◽  
Vol 9 (9) ◽  
pp. 609-613 ◽  
Author(s):  
Ingolf Krause ◽  
Johann Buchberger ◽  
Günther Weiß ◽  
Monika Pflügler ◽  
Henning Klostermeyer
Keyword(s):  
High Resolution ◽  
Isoelectric Focusing ◽  
Genetic Variant ◽  
Ph Gradients ◽  
Carrier Ampholytes

An improved capillary isoelectric focusing-mass spectrometry method for high-resolution characterization of monoclonal antibody charge variants

2022 ◽  
Author(s):  
Tian Xu ◽  
Linjie Han ◽  
Alayna M. George Thompson ◽  
Liangliang Sun
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
High Resolution ◽  
Isoelectric Focusing ◽  
Capillary Isoelectric Focusing ◽  
Spectrometry Method ◽  
Mass Spectrometry Method ◽  
Charge Variants

Routine and high-resolution characterization of monoclonal antibody (mAb) charge variants is vital for controlling mAb quality as therapeutics. Capillary isoelectric focusing-mass spectrometry (cIEF-MS) has emerged as a powerful tool for...

Characterization of synthetic carrier ampholytes for isoelectric focusing

1975 ◽  
Vol 109 (2) ◽  
pp. 341-356 ◽  
Author(s):  
Pier Giorgio Righetti ◽  
Mariangela Pagani ◽  
Elisabetta Gianazza
Keyword(s):  
Isoelectric Focusing ◽  
Carrier Ampholytes

Chromatofocusing and isoelectric focusing in immobilized pH gradients compared for characterization of human hemoglobin variants.

1989 ◽  
Vol 35 (3) ◽  
pp. 425-430 ◽  
Author(s):  
R Paleari ◽  
C Arcelloni ◽  
R Paroni ◽  
I Fermo ◽  
A Mosca
Keyword(s):  
Los Angeles ◽  
Isoelectric Focusing ◽  
Ph Gradient ◽  
Resolving Power ◽  
Human Hemoglobin ◽  
Ph Gradients ◽  
Hemoglobin Variants

Abstract We compared the performance of two highly resolving methods, chromatofocusing (CRF) and isoelectric focusing in immobilized pH gradients (IPGF), for the separation of human hemoglobin variants. Lysates containing 13 different hemoglobins, including variants of clinical and geographical importance, and four electrophoretically "silent" variants (Hb Brockton, Hb Cheverly, Hb Köln, and Hb Waco) were analyzed. Both techniques showed a good intrarun precision (CV = 0.87% for CRF, 0.27% for IPGF) and high and similar resolving power (0.010 pH units, with the pH gradients used in this work). The use of an ultranarrow IPGF range (pH 7.15-7.35; pH gradient = 0.019 pH/cm) allowed the resolution between Hb Brockton, Hb Köln, and Hb A. In some cases (Hb D-Los Angeles, Hb F, Hb Waco), the variants were separated from Hb A in different orders, depending on which technique was used, probably because of the different analytical principles of the two methods. As a second-level test, both procedures are informative for characterization of human hemoglobin variants.

Characterization of proteins by sequential isoelectric focusing on immobilized pH gradients and electrospray mass spectrometry

1995 ◽  
Vol 16 (1) ◽  
pp. 1381-1384 ◽  
Author(s):  
Umberto Breme ◽  
Jer??me Breton ◽  
Carlo Visco ◽  
Gaetano Orsini ◽  
Pier Giorgio Righetti
Keyword(s):  
Mass Spectrometry ◽  
Isoelectric Focusing ◽  
Electrospray Mass Spectrometry ◽  
Ph Gradients

scholarly journals A Mini Review on Capillary Isoelectric Focusing-Mass Spectrometry for Top-Down Proteomics

2021 ◽  
Vol 9 ◽  
Author(s):  
Tian Xu ◽  
Liangliang Sun
Keyword(s):  
Mass Spectrometry ◽  
High Resolution ◽  
Isoelectric Focusing ◽  
Large Scale ◽  
Protein Characterization ◽  
Capillary Isoelectric Focusing ◽  
Top Down ◽  
Esi Ms ◽  
Highly Sensitive

Mass spectrometry (MS)-based top-down proteomics (TDP) requires high-resolution separation of proteoforms before electrospray ionization (ESI)-MS and tandem mass spectrometry (MS/MS). Capillary isoelectric focusing (cIEF)-ESI-MS and MS/MS could be an ideal method for TDP because cIEF can enable separation of proteoforms based on their isoelectric points (pIs) with ultra-high resolution. cIEF-ESI-MS has been well-recognized for protein characterization since 1990s. However, the widespread adoption of cIEF-MS for the characterization of proteoforms had been impeded by several technical challenges, including the lack of highly sensitive and robust ESI interface for coupling cIEF to MS, ESI suppression of analytes from ampholytes, and the requirement of manual operations. In this mini review, we summarize the technical improvements of cIEF-ESI-MS for characterizing proteoforms and highlight some recent applications to hydrophobic proteins, urinary albumin variants, charge variants of monoclonal antibodies, and large-scale TDP of complex proteomes.

Sign in / Sign up

Export Citation Format

Share Document