Chromatographic Studies of Protein-Based Chiral Separations.

Sandya Beeram,Cong Bi,David Hage,Xiwei Zheng,Shiden Azaria,Zhao Li

doi:10.3390/separations3030027

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.

Highlights

The use of molecular chirality can be traced back to the mid-1800s in the work by LouisPasteur with tartaric acid [1,2,3]
Significant effort has been aimed at creating chiral stationary phases (CSPs) that can be used in chromatography for the separation and analysis of chiral compounds [3,4,5,6,7,8,9]
One type of Acid glycoprotein (AGP)-based CSP was prepared by mildly oxidizing the carbohydrate chains on AGP and using these modified residues to immobilize this glycoprotein onto hydrazide-activated silica [34,58]

Summary

Introduction

The use of molecular chirality can be traced back to the mid-1800s in the work by Louis. Several methods have been employed for the chiral separation of drugs and amino acids for chemical analysis These methods have included gas chromatography, thin layer chromatography, high-performance liquid chromatography (HPLC), supercritical fluid chromatography, and capillary electrophoresis [6,9,16,17,18]. The properties and applications for several specific types of protein-based CSPs will be examined This will include CSPs that employ various types of serum transport proteins and enzymes, as well as of CSP will be discussed, along with information that has been obtained through chromatography on these materials and their ability to bind and separate chiral agents

Zonal Elution

Frontal Analysis

Kinetic Studies

Preparation of Protein-Based

Supports Used in Protein-Based CSPs

Development and Evaluation of Protein Immobilization Methods

Physical Adsorption

Covalent Immobilization

Encapsulation and Entrapment

Serum Transport Proteins

Human Serum Albumin and Bovine Serum Albumin

Chiral separation separation of D