Analysis of Biological Interactions by Affinity Chromatography: Clinical and Pharmaceutical Applications

Abstract

BACKGROUND The interactions between biochemical and chemical agents in the body are important in many clinical processes. Affinity chromatography and high-performance affinity chromatography (HPAC), in which a column contains an immobilized biologically-related binding agent, are two methods that can be used to study these interactions. CONTENT This review looks at various approaches that can be used in affinity chromatography and HPAC to characterize the strength or rate of a biological interaction, the number and types of sites that are involved in this process, and the interactions between multiple solutes for the same binding agent. A number of applications for these methods are examined, with an emphasis on recent developments and high-performance affinity methods. These applications include the use of these techniques for fundamental studies of biological interactions, high-throughput screening of drugs, work with modified proteins, tools for personalized medicine, and studies of drug-drug competition for a common binding agent. SUMMARY The wide range of formats and detection methods that can be used with affinity chromatography and HPAC for examining biological interactions makes these tools attractive for various clinical and pharmaceutical applications. Future directions in the development of small-scale columns and the coupling of these methods with other techniques, such as mass spectrometry or other separation methods, should continue to increase the flexibility and ease with which these approaches can be used in work involving clinical or pharmaceutical samples.

Challenges in Separations of Proteins and Small Biomolecules and the Role of Modern Mass Spectroscopy Tools for Solving Them, as Well as Bypassing Them, in Structural Analytical Studies of Complex Biomolecular Mixtures

“…The most popular method of protein purification today is affinity purification [32,33], using antibodies [34], synthetic peptides [29] or other affinity reagents [35]. Immunoprecipitation is highly popular today, due to the significantly increased industrial supply of antibodies, particularly against human and mouse proteins.…”

Investigation of the enantioselective interaction between selected drug enantiomers and human serum albumin by mobility shift‐affinity capillary electrophoresis

“…Molecular interactions in the biological system are mostly related to the availability of human serum proteins. HSA and α-acid glycoprotein are the most important transporter proteins due to their ability to bind to a wide variety of drugs [4][5][6]. HSA is a single-chain, nonglycosylated polypeptide containing 585 amino acid residues with a molecular mass of 66.5 kDa.…”

Solid-Supported Proteins in the Liquid Chromatography Domain to Probe Ligand-Target Interactions

“…The use of solid-supported proteins to evaluate protein-protein interactions and to purify proteins is a well-founded tool (Muronetz et al, 2001; Perret and Boschetti, 2018). Interactions between proteins and small molecules have also been well-explored (de Moraes et al, 2014a; Zheng et al, 2014; Hage, 2017). More recently, solid-supported proteins have been employed not only to assess these interactions, but mainly as a strategy to isolate small molecules from complex combinatory libraries (Forsberg and Brennan, 2014; Zhuo et al, 2016; Vanzolini et al, 2018a; Wang L. et al, 2018).…”

“…Terms such as weak affinity chromatography (WAC) (Meiby et al, 2013; Singh P. et al, 2017; Ohlson and Duong-Thi, 2018; Lecas et al, 2019), high-performance affinity chromatography (HPAC) (Hage, 2017; Li Z. et al, 2017; Beeram et al, 2018; Zhang C. et al, 2018), high-performance liquid affinity chromatography (HPLAC) (Zheng et al, 2014), biointeraction chromatography (Wainer, 2004), affinity monolith chromatography (AMC) (Lecas et al, 2019), and cellular chromatography (CC) (Ciesla et al, 2016; Xu et al, 2019) have been largely and correctly employed, but all of them could be well-settled in the general term bioaffinity chromatography, which has been introduced to differentiate them from the classic affinity chromatography (de Moraes et al, 2016).…”