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mAB Analytic via Protein A Columns
Analysis of Immunoglobulins (Antibodies)
Affinity Chromatography with Protein A
Immunoglobulins (antibodies) are proteins that are produced in humans and animals in response to certain substances (antigens). These include immunoglobulin G (IgG) and monoclonal antibodies (mAb's), which are of increasing importance in today's medical research and applications.
The analysis or purification of antibodies is often carried out by protein A affinity chromatography, as protein A has the ability to bind many of the immunoglobulins reversibly. Under physiological conditions, protein A and its recombinant derivatives bind to the Fc part of the antibodies and their aggregates. The immunoglobulins are eluted by lowering the pH value.
The main field of application of protein A affinity chromatography media is capturing* during the purification process of antibodies. In industry, this is carried out in large self-packed columns. Protein A HPLC columns are used, among other things, to determine the concentration of mAb's during cell line selection or in the upstream of bioprocess optimisation. Furthermore, small amounts of immunoglobulins can also be purified for analytical purposes.
[English:] Figure 1: [English:] Schematic illustration of an antibody
Applications
Affinity chromatography Purification of IgG with Tosoh TSKgel Protein A-5PW
Peak identities
1. impurities 2. IgG
Test conditions
Column: TSKgel Protein A-5PW, 35x4.6mm, 20µm
Mobile phase binding: 20 mmol/L sodium phosphate, pH 7.4
Mobile phase elution: 20 mmol/L sodium phosphate, pH 2.5
Gradient: 0-0.5 min binding buffer, 0.5-1.1 min elution buffer, 1.1-2.0 min binding buffer
Flow rate: 2 mL/min
Detection. UV, 280 nm
Sample: 20 µL CHO cell culture supernatant with IgG (0.5 mg/mL)
Downloads
Manufacturer overview
Sepax
HPLC - Affinity chromatography:
- MAbPurix
Thermo Fisher
HPLC - affinity chromatography:
- MAbPac
Tosoh
HPLC - Affinity chromatography:
- TSKgel Protein A-5PW, 20µm
- TOYOPEARL AF-rProtein A HC-650F
- Tosoh TOYOPEARL AR-rProtein A Increased productivity for downstream processin of biotherapeutics Applications
- Tosoh TSKgel Protein A-5PW Reproducible and Robust Antibody Titer Analysis Applications
- Tosoh TSKgel Protein A-5PW Fast mAb Titer Determination Applications
- TOSOH Comparison of Toyopearl AF-rProtein a HC-650F binding capacity at various bed heights and constant linear velocity
- Tosoh TOYOPEARL AF-rProtein A HC-650F Improve Performance of the Protein A Capturing Step Applications
- Tosoh TOYOPEARL AR-rProtein A HC-650F Protein A chromatography - The process economics driver in mab manufacturing Applications
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