A REVIEW ON IMMUNOAFFINITY CHROMATOGRAPHY

Affinity chromatography is a type of liquid chromatography that makes use of biological-like interactions for the separation and specific analysis of sample components. This review describes the basic principles of immunoaffinity chromatography and examines its use in the testing of clinical samples, with an emphasis on HPLC based methods. Some traditional applications of this approach include the use of baronage, lectin, protein A or protein G, and immune affinity supports for the direct quantification of solutes. Newer techniques that use antibody-based columns for on- or off-line sample extraction are examined in detail, as are methods that use affinity chromatography in combination with other analytical methods, such as reversed-phase liquid chromatography, gas chromatography, and capillary electrophoresis. Indirect analyte detection methods are also described in which immune affinity chromatography is used to perform flow-based immunoassays. Other applications that are reviewed include affinity-based chiral separations and the use of affinity chromatography for the study of drug or hormone interactions with binding proteins. Some areas of possible future developments are then considered, such as tandem affinity methods and the use of synthetic dyes, immobilized metal ions, molecular imprints, or aptamers as affinity ligands for clinical analytes. Immunoaffinity chromatography is the process in which the binding affinity of an antigen to a parent antibody is utilized as a basis of separation. Owing to their avidity and specificity, monoclonal antibodies have become indispensible for both protein characterization and purification. The article describes the basic procedure of immunoaffinity chromatography. The support matrix upon which the antibody is immobilized and the activation chemistry used to couple the antibody to the matrix affect the immunosorbent performance. Support matrices available for Immunoaffinity chromatography and their activation chemistries including the recent advances have been reviewed. The paper also discusses the developments in the applications of this technique in analysis as well as extraction.