Articles by Agilent Technologies - BioPharm International

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Articles by Agilent Technologies

Precise Determination of Protein Molecular Weight

Sep 24, 2013

This Application Note shows the determination of molecular weight, size, and relative concentration of protein samples after SEC with the Agilent 1260 Infinity Bio-inert Quaternary LC System with Multi-Detector GPC/SEC.

High-resolution Analysis of Charge Heterogeneity in Monoclonal Antibodies Using pH-gradient Cation Exchange Chromatography

Jun 25, 2013

The authors describe pH-based separation of acidic and basic charge variants for mAbs using the Agilent 1260 Infinity Bio-inert Quaternary LC System and an Agilent BioMAb PEEK 4.6 x 250 mm IEX column, featuring a special particle coating designed for charge-based separation of MAbs.

Characterization of Glycosylation in the Fc Region of Therapeutic Recombinant Monoclonal Antibody

May 28, 2013

In this note, we describe a fast and efficient liquid chromatography-TOF mass spectrometry approach to characterize an IgG derived from CHO cells. An Agilent Poroshell 300SB-C3, 5 um superficially porous IgG1 column was used to obtain accurate glycoform masses of an intact IgG1, with a subsequent analysis of papain-digeste3d IgG1 to obtain site-specific glycosylation profile information of the Fc region for further characterization of IgG1 heterogeneity. Additionally, a subsequent mAb-glyco chip LC/MS analysis was used to build a glycan accurate mass database for additional validation of the glycans identified in the TOF mass analysis.

Analysis of Polyethylene Glycol (PEG) and a Mono and Di-PEGylated Therapeutic Protein Using HPLC and Q-TOF Mass Spectrometry

May 28, 2013

A method using LC/MS is described for improving the analysis PEGylated peptides and proteins using the post-column addition of a charge stripping agent to obtain simpler mass spectra for detailed structural characterization.

Analysis of Erythropoietin Glycopeptides and Glycoforms by HILIC and RP Chromatography

May 28, 2013

Reversed-phase UHPLC/HPLC is routinely used for peptide mapping, but if the digest contains hydrophilic peptides, such as glycopeptides, valuable information can be missed. This note demonstrates peptide mapping of digested glycoprotein erythropoietin (EPO) protein using an Agilent ZORBAX Rapid Resolution High Definition 300-HILIC 1.8 ?m LC column and a TOF MS. Taking advantage of the high organic solvent system of the mobile phase for HILIC (hydrophilic interaction chromatography), the glycopeptides are effectively dissolved, retained and separated by the HILIC column. This app note includes the separation, sequence coverage and comparison of reversed-phase and HILIC data to demonstrate the utility of HILIC as an orthogonal and complementary approach to RPLC/MS for peptide analysis.

Protein Separation with pH Gradients Using Composite Buffer Systems

Apr 30, 2013

Tips and tricks on how to correctly apply the Agilent Buffer Advisor Software for the generation of pH gradients over a wide range using composite buffer systems.

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