Capillary Isoelectric Focusing-Mass Spectrometry Method for the Separation and Online Characterization of Intact Monoclonal Antibody Charge Variants
نویسندگان
چکیده
منابع مشابه
Monoclonal Antibody Charge Heterogeneity Analysis by Capillary Isoelectric Focusing on the Agilent 7100 Capillary Electrophoresis System
Due to the importance of monoclonal antibodies (mAb) as therapeutic agents, there is a growing demand for high resolution analytical methods to characterize these complex molecules. One important property of an mAb is its charge state that may change during and after manufacturing. A very useful technique to measure mAb charge heterogeneity is capillary isoelectric focusing (cIEF). This Applica...
متن کاملCapillary isoelectric focusing-mass spectrometry for shotgun approach in proteomics.
We report on capillary isoelectric focusing-mass spectrometry (CIEF-MS) of complex peptide mixtures in the absence of carrier ampholytes. Furthermore, the use of low concentrations of carrier ampholytes as mere spacers is investigated. Carrier ampholytes are complex mixtures of amphoteric compounds with high buffering capacity. Since all peptides are amphoteric compounds by themselves, the use ...
متن کاملOn-line capillary isoelectric focusing hyphenated to native electrospray ionization mass spectrometry for the characterization of interferon-γ and variants.
The on-line hyphenation of Capillary IsoElectric Focusing (CIEF) with ElectroSpray Ionization Mass Spectrometry (ESI/MS) has been carried out in a non-denaturing detection mode at the CIEF-MS interface. This CIEF-MS coupling methodology relied on the use of 40% glycerol-water medium as anti-convective agent in the CE capillary and the addition of 10 mM ammonium acetate buffer, pH 5, as a volati...
متن کاملImaged Capillary Isoelectric Focusing for Charge-Variant Analysis of Biopharmaceuticals
level: interMediAte Analyzing charge variants of therapeutic proteins is critical for characterizing and monitoring quality attributes of antibodies. Charge variants include deamidation, formation of N-terminal pyroglutamate, aggregation, isomerization, sialylated glycans, antibody fragmentation, and glycation at the lysine residues. In some cases, such changes affect binding, biological activi...
متن کاملComparison of Different Brands of Carrier Ampholytes for Monoclonal Antibody Charge Heterogeneity Analysis by Capillary Isoelectric Focusing
This Application Note compares the performance of four commercially available brands of wide-range pH 3 to 10 carrier ampholytes, Pharmalyte, Servalyt, HR, and SH AESlyte, for high-resolution capillary isoelectric focusing on fluorocarbon-coated capillaries. The carrier ampholytes-specific background was increased for Servalyt, and slightly increased for SH AESlyte in comparison to Pharmalyte a...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Analytical Chemistry
سال: 2018
ISSN: 0003-2700,1520-6882
DOI: 10.1021/acs.analchem.7b04608