Antibody-based separation methods, such as immunoaffinity chromatography (IAC), are powerful purification and isolation techniques. Antibodies isolated using these techniques have proven highly efficient in applications ranging from clinical diagnostics to environmental monitoring. Immunoaffinity chromatography is an efficient antibody separation method which exploits the binding efficiency of a ligand to an antibody. Essential to the successful design of any IAC platform is the optimization of critical experimental parameters such as (a) the biological affinity pair, (b) the matrix support, (c) the immobilization coupling chemistry, and (d) the effective elution conditions. These elements and the practicalities of their use are discussed in detail in this review. At the core of all IAC platforms is the high affinity interactions between antibodies and their related ligands; hence, this review entails a brief introduction to the generation of antibodies for use in immunoaffinity chromatography and also provides specific examples of their potential applications.
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