Affinity Chromatography: A Review of Clinical Applications

Abstract

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 affinity 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 boronate, lectin, protein A or protein G, and immunoaffinity 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 immunoaffinity 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.

High-level expression of active recombinant ubiquitin carboxyl-terminal hydrolase of Drosophila melanogaster in Pichia pastoris

“…Purification of His-tagged proteins is based on the chelating metal ions as affinity ligands. The use of short histidine stretches or His-tags, typically as affinity tags at either the N-terminus or C-terminus, enables the purification of the desired protein from the crude extract in a single step by IMAC (immobilized metal ion affinity chromatography) [40][41][42]. The most widely used IMAC supports are either nickel nitrilotriacetic acid (Ni-NTA) resins or different chelating Sepharose matrices.…”

Maßgeschneiderte Adsorbentien im Anwendungsspektrum Bio-, Medizin- und Umwelttechnik

“…B. auch für Nucleinsäuren zur Anwendung in Biochips oder ¹Lab on a chipª-Konzepten. Auûer in der Biotechnologie wird die Affinitätschromatographie in der klinischen Diagnostik [74] eingesetzt, z. B. auf der Basis von immobilisierten Lectinen bei der krankheitsbegleitenden Diagnostik alkoholbedingter Lebererkrankungen [75] oder Prostatakrebsleiden [76].…”

mRNA display selection and solid‐phase synthesis of Fc‐binding cyclic peptide affinity ligands

“…Consequently, replacement of the highly expensive Protein A/G media for mAb purification is an important goal (DiMasi and Grabowski, 2007;Ramberg, 2002). To this end, several novel and robust synthetic affinity ligands have been proposed (Clonis, 2006;Hage, 1999;Low et al, 2007;Newcombe et al, 2005;Roque et al, 2004Roque et al, , 2007 and among these, peptides are a growing class (Kabir, 2002;Lund et al, 2012;Yang et al, 2008Yang et al, , 2010. In particular, recent studies have drawn attention to cyclic peptides as a new class of molecules with great potential (Corti et al, 2011;Frecer et al, 2004;Heinis et al, 2009;Li and Cho, 2010;Li et al, 2002;Mantey et al, 2001;Misumi et al, 2003;Neumann and Neumann-Staubitz, 2010;Strohl and Knight, 2009;Vlieghe et al, 2010;Wels et al, 2005).…”