Related Experiment Video
Updated: Jul 5, 2026
11:01
Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Published on: January 18, 2020
Purification of immunoglobulin G
1Lancaster University, Lancaster, United Kingdom.
Current Protocols in Immunology
|April 25, 2008
Summary
Purify immunoglobulin G (IgG) using cost-effective ammonium sulfate precipitation and size-exclusion chromatography. Alternative methods like protein A/G or anti-rat antibody affinity chromatography and ion-exchange chromatography are also discussed for antibody purification.
Area of Science:
- Biochemistry
- Immunology
- Analytical Chemistry
Background:
- Immunoglobulin G (IgG) purification is crucial for research and therapeutic applications.
- Various chromatographic techniques exist for antibody purification, each with advantages and limitations.
Purpose of the Study:
- To describe and compare different methods for purifying IgG antibodies.
- To provide protocols for specific antibody purification strategies.
Main Methods:
- Ammonium sulfate precipitation followed by size-exclusion (SE) chromatography.
- Protein A and Protein G affinity chromatography.
- Anti-rat antibody affinity chromatography.
- Ion-exchange (IEX) chromatography.
Main Results:
- Ammonium sulfate precipitation with SE chromatography is the most economical IgG purification method.
- Protein A/G affinity chromatography offers speed but limited subclass effectiveness for rat antibodies.
- Anti-rat antibody affinity chromatography provides a specific protocol for rat antibody purification.
- IEX chromatography is suitable for purifying intact antibodies and fragments.
Conclusions:
- Multiple validated methods exist for IgG purification, catering to different needs regarding cost, speed, and antibody type.
- The choice of purification method depends on factors like antibody subclass, species, and required purity.
- Detailed protocols are provided for effective antibody purification strategies.
Related Concept Videos
01:20Immunoprecipitation
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
01:29Types Of Column Chromatography
The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
Gel Filtration Chromatography
When the...