Separation of antigens and antibodies by immunoaffinity chromatography

Abstract

Context: Affinity chromatography is an efficient antibody, antigen and protein separation method based on the interaction between specific immobilized ligands and target antibody, antigen, and so on. Populations of available ligands can be used to separate antibodies or their Fab fragments. Similarly, antigens can be isolated by immunoaffinity chromatography (IAC) on immobilized antibodies of low affinity.

Objective: This review describes the advantages, the applications, as well as the drawbacks, of IAC in the separation and purification of antibodies and antigens.

Methods: The present review discussed all types of purification and isolation of antibodies and antigens by IAC, including purification of antibodies using immobilized and synthetic mimic proteins A, G and L; isolation of Fab fragments of antibodies; separation of antibodies against different antigen forms; isolation of antigens by immobilized antibodies and so on. These methods come from over 60 references compiled from all major databases.

Results: Purification of antigens with antibodies should choose low-affinity antibodies to avoid denaturation of most proteins. Concern for cost and safety, prompted research activities focused on novel synthetic ligands with improved properties such as lower cost, avoidance of the risk of contamination associated with natural ligands of human or animal origin to isolate antibodies and antigens.

Conclusion: It is anticipated that the improvements of IAC will have impact not only on large-scale production of antibodies but also on the generation of new affinity-based methods for the increasing number of proteins and antibody derivatives available by protein engineering and the proteomics revolution.

Keywords::

• Affinity chromatography
• isolation
• purification
• immobilized proteins
• synthetic peptide ligand

Acknowledgment

We thank Miss Trista Talbot (Department of Chemistry, Brandeis University) for language editing of the manuscript.

Declaration of interest

The authors report no declarations of interest.