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Human Auto-IgG Purification from High Volume Serum Sample by Protein G Affinity Purification
Serena Sensi1, Andreas Goebel1
1Dept of Musculoskeletal and Ageing Science, University of Liverpool, Liverpool, United Kingdom.
Bio-Protocol
|December 23, 2022
Summary
This study details a protein G affinity chromatography protocol for purifying high-quality human immunoglobulin G (IgG) from serum. This method is efficient for large-scale purification, crucial for applications like animal immunization.
Area of Science:
- Biochemistry
- Immunology
- Protein Chemistry
Background:
- Immunoglobulins (IgG) are vital proteins for immune response.
- Purified human IgG is essential for research, including hybridoma production and animal immunization.
- Existing purification methods have limitations in specificity and scalability.
Purpose of the Study:
- To provide a detailed, validated protocol for high-quality human IgG purification from serum.
- To optimize IgG purification using protein G affinity chromatography for research applications.
- To enable efficient purification of large quantities of human IgG.
Main Methods:
- Affinity chromatography using protein G immobilized on Sepharose beads.
- Step-wise protocol including column preparation, serum loading, washing, and elution.
- Post-purification steps: dialysis for buffer exchange and concentration.
Main Results:
- Successful purification of high-quality human IgG.
- Protein G demonstrates high affinity for human IgG, ensuring specific binding.
- The protocol is effective for purifying large amounts of IgG.
Conclusions:
- The described protein G affinity chromatography protocol offers a reliable and scalable method for human IgG purification.
- This method is particularly suitable for preparing IgG for animal immunization studies.
- The protocol is validated and applicable for demanding research needs.
