Agilent 3100/2100 Antibody Charge Variant Analysis Application Note

This document describes a method for monitoring antibody charge variants using Agilent 3100 OFFGEL fractionation and high sensitivity protein detection with the Agilent 2100 Bioanalyzer. This method can be used for quality control in the production of monoclonal antibodies in the biopharmaceutical industry.

Brand: Agilent
Category: Other
Document type: Application Note
Language: English
Pages: 4
File format: PDF
File size: 771 KB
Published: 12 June, 2012
Updated: 28 September, 2023
Model: 2100 Bioanalyzer
MD5 checksum: 13C3BF533DF3A7EB9D0078654F570A0D
PDF: Agilent 3100/2100 Antibody Charge Variant Analysis Application Note PDF

About this manual

This document details a two-dimensional separation method for analyzing monoclonal antibody (mAb) charge heterogeneity. The first dimension utilizes Agilent 3100 OFFGEL Fractionation, an isoelectric focusing technique, to separate proteins based on charge. The second dimension employs the Agilent 2100 Bioanalyzer with the High Sensitivity Protein 250 Assay to separate and detect the resulting fractions by size.

What's inside

Abstract
Introduction
Materials and Methods
Results and Discussion
Figure 1
Figure 2

Specifications

Resolution	0.1 pH
Sample Load	100 pg
Molecular Weight Difference	4 kDa
Variant Percentage	10%
Detection Range	picogram range

Frequently asked questions

What technique is used for the first dimension of separation?

Agilent 3100 OFFGEL Fractionation, an isoelectric focusing technique.

Which assay is used for the second dimension analysis?

The Agilent High Sensitivity Protein 250 Assay is used for size separation and detection.

What is the purpose of adding Tween-20 to the buffer?

Tween-20 is added to enhance the solubility of the proteins during focusing.

Download PDF (771 KB)