Purification of Antibodies Using Affinity Chromatography

Area of Science:

• Biochemistry
• Immunology
• Chromatography

Background:

• Affinity chromatography isolates target molecules from complex mixtures using specific binding interactions.
• The discovery of proteins A, G, and L has made immuno-affinity chromatography a standard for antibody purification.
• Antibodies are crucial for applications like immunodiagnostics.

Purpose of the Study:

• To provide researchers with basic techniques for affinity chromatography-based purification of antibodies.
• To guide the selection of appropriate affinity ligands for antibody isolation.
• To detail the application of proteins A and G in antibody purification.

Main Methods:

• Exploitation of specific interactions between affinity pairs (e.g., antibody-antigen, ligand-receptor).
• Immuno-affinity chromatography utilizing immobilized proteins A, G, or L.
• Purification of monoclonal, polyclonal, and recombinant antibodies.

Main Results:

• Established immuno-affinity chromatography as the standard for antibody purification.
• Demonstrated the utility of proteins A and G for antibody isolation.
• Provided tables to aid researchers in selecting the optimal protein ligand.

Conclusions:

• Affinity chromatography is a versatile technique for purifying various analytes, especially antibodies.
• Proteins A and G are effective ligands for antibody purification in immuno-affinity chromatography.
• This chapter serves as a practical resource for researchers purifying antibodies.