A proof-of-concept analysis of carbohydrate-deficient transferrin by imaged capillary isoelectric focusing and in-capillary immunodetection

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Imaged 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...

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Capillary Isoelectric Focusing

Beale SC (1998) Capillary electrophoresis. Analytical Chemistry 70: 279. (This journal prints every two years a review on capillary electrophoresis. The article is written by an expert in the Reld and covers more or less all developments in CE including CGE of the preceding years.) Grossmann PD and Colburn JC (eds) (1996) Capillary Electrophoresis: Theory and Practice. San Diego: Academic Press...

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Determination of carbohydrate-deficient transferrin using capillary zone electrophoresis.

BACKGROUND Current methods for carbohydrate-deficient transferrin (CDT) often suffer from low precision, complexity, or risk of false positives attributable to genetic variants. In this study, a new capillary zone electrophoresis (CZE) method for CDT was developed. METHODS CZE was performed on a P/ACE 5000 using fused-silica capillaries [50 microm (i.d.) x 47 cm] and the CEOFIX CDT buffer sys...

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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...

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Quantification of hemoglobin variants by capillary isoelectric focusing.

Capillary isoelectric focusing (cIEF) was used to identify and quantify major and minor hemoglobin (Hb) variants. Whole blood (approximately 10 microL required) hemolysate was analyzed with a commercial instrument equipped with a 50 microns (i.d.) x 27 cm coated capillary filled with 20 g/L ampholytes (pH 6-8) in 4 g/L methylcellulose (MC). Cathode and anode solutions were 20 mol/L NaOH and 100...

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ژورنال

عنوان ژورنال: BioTechniques

سال: 2020

ISSN: 0736-6205,1940-9818

DOI: 10.2144/btn-2019-0058