Chromatographic Studies of Protein-Based Chiral Separations

Abstract

The development of separation methods for the analysis and resolution of chiral drugs and solutes has been an area of ongoing interest in pharmaceutical research. The use of proteins as chiral binding agents in high-performance liquid chromatography (HPLC) has been an approach that has received particular attention in such work. This report provides an overview of proteins that have been used as binding agents to create chiral stationary phases (CSPs) and in the use of chromatographic methods to study these materials and protein-based chiral separations. The supports and methods that have been employed to prepare protein-based CSPs will also be discussed and compared. Specific types of CSPs that are considered include those that employ serum transport proteins (e.g., human serum albumin, bovine serum albumin, and alpha1-acid glycoprotein), enzymes (e.g., penicillin G acylase, cellobiohydrolases, and α-chymotrypsin) or other types of proteins (e.g., ovomucoid, antibodies, and avidin or streptavidin). The properties and applications for each type of protein and CSP will also be discussed in terms of their use in chromatography and chiral separations.

Covalent Organic Frameworks with Chirality Enriched by Biomolecules for Efficient Chiral Separation

“…[1] Theseparation of racemic compounds has long been of great significance,especially for pharmaceutical and medicinal applications, [2,3] which has spurred continuous interest in the exploration of new materials/approaches for efficient separation of enantiomers. [8][9][10] Biomolecules such as enzymes that are created by nature, [11] can well discriminate enantiomers owing to their natural conformations composed of chiral subunits (that is,amino acids) as well as amphiphilic and zwitterionic features capable of providing specific interactions.T his makes them appealing for chiral separation particularly as chiral stationary phases (CSPs) in chromatography if they can be immobilized on some solidstate materials.H erein, we contribute ag eneral approach to immobilize biomolecules into an ew class of solid-state materials,c ovalent organic frameworks (COFs), and the afforded biomolecules&COFs can serve as versatile and highly efficient CSPs towards various racemates in both normal-phase and reverse-phase high-performance liquid chromatography. [8][9][10] Biomolecules such as enzymes that are created by nature, [11] can well discriminate enantiomers owing to their natural conformations composed of chiral subunits (that is,amino acids) as well as amphiphilic and zwitterionic features capable of providing specific interactions.T his makes them appealing for chiral separation particularly as chiral stationary phases (CSPs) in chromatography if they can be immobilized on some solidstate materials.H erein, we contribute ag eneral approach to immobilize biomolecules into an ew class of solid-state materials,c ovalent organic frameworks (COFs), and the afforded biomolecules&COFs can serve as versatile and highly efficient CSPs towards various racemates in both normal-phase and reverse-phase high-performance liquid chromatography.…”

“…Thep eptide&COF 1 only exhibited separation effect towards some of the tested racemates with low separation efficiency( Supporting Information, Figures S24, S25). [11,44] Therefore,surface-enhanced Raman scattering (SERS) spectra were collected to probe the specific interactions between racemic substrates and lysozyme (tryptophan was selected as ar epresentative substrate). Lysine&COF 1 failed to separate any of the tested racemates (Supporting Information, Figure S28), which highlighted the importance of higher order structure of biomolecules (for example,l ysozyme) in chiral recognition and resolution.…”

“…[4][5][6][7] To achieve efficient separation of enantiomers,i tis essential the material possess both strong chiral environment and preferable binding ability through some specific interactions (for example,h ydrogen bonding,v an der Waals interactions,and electrostatic interactions). [8][9][10] Biomolecules such as enzymes that are created by nature, [11] can well discriminate enantiomers owing to their natural conformations composed of chiral subunits (that is,amino acids) as well as amphiphilic and zwitterionic features capable of providing specific interactions.T his makes them appealing for chiral separation particularly as chiral stationary phases (CSPs) in chromatography if they can be immobilized on some solidstate materials.H erein, we contribute ag eneral approach to immobilize biomolecules into an ew class of solid-state materials,c ovalent organic frameworks (COFs), and the afforded biomolecules&COFs can serve as versatile and highly efficient CSPs towards various racemates in both normal-phase and reverse-phase high-performance liquid chromatography.Emerging as an ew class of crystalline solid-state materials,COFs feature high surface area, low mass density,tunable pore size,high stability,and easily tailored functionality, [12][13][14] which means they hold promise for applications in many fields such as gas storage, [15] photoelectricity, [16] catalysis, [17][18][19] environmental remediation, [20] drug delivery, [21] and functional devices. [22] Thedevelopment of COFs for chiral separation is still in the infancy stage, [23][24][25] primarily relying on the construction of chiral COFs based on chiral monomers.O n the basis of our recent success in immobilizing enzymes into COFs, [26] in this work we present an alternative strategy to introduce chirality into COFs by covalently anchoring aseries of biomolecules,such as amino acids,peptides,and enzymes, onto the channel walls of achiral COFs to form biomolecu-les&COFs (Scheme 1).…”

Fundamental Developments of Chiral Phase Chromatography in Connection with Enantioselective Synthesis of β‐Amino Acids

“…(b) Sorbents based on optically active polymers, which can be synthetic such as the molecularly imprinted polymers or obtained from natural sources, e. g. polysaccharides derivatives, or protein based CSP …”

Chiral Recognition by DNA-Immobilized TLC Plate

“…These biopolymers have played an important role in biological processes. In the material science fields, these biopolymers with a chiral property have been used as a separation material of chiral substances [1], a sensing material of chiral drugs [6], a chiral catalyst for chemical reactions [7], or a nonlinear optical material [8]. Similarly, the double-stranded DNA, one of the most important biopolymers in living things, consists of two antiparallel polynucleotide strands intertwined with one another to form a right-handed double helix [9,10].…”