High performance affinity chromatography and related separation methods for the analysis of biological and pharmaceutical agents

Abstract

The last few decades have witnessed the development of many high-performance separation methods that use biologically related binding agents. The combination of HPLC with these binding agents results in a technique known as high performance affinity chromatography (HPAC). This review will discuss the general principles of HPAC and related techniques, with an emphasis on their use for the analysis of biological compounds and pharmaceutical agents. Various types of binding agents for these methods will be considered, including antibodies, immunoglobulin-binding proteins, aptamers, enzymes, lectins, transport proteins, lipids, and carbohydrates. Formats that will be discussed for these methods range from the direct detection of an analyte to indirect detection based on chromatographic immunoassays, as well as schemes based on analyte extraction or depletion, post-column detection, and multi-column systems. The use of biological agents in HPLC for chiral separations will also be considered, along with the use of HPAC as a tool to screen or study biological interactions. Various examples will be presented to illustrate these approaches and their applications in fields such as biochemistry, clinical chemistry, and pharmaceutical research.

Development of a microcolumn one-site immunometric assay for a protein biomarker: Analysis of alpha1-acid glycoprotein

“…Also, F will be greater than 0.90 when f [Ab*] T is at least 10-fold larger than K d (i.e., more than 90% of the analyte is present in the complex Ab*•A after incubation). It is known that a typical polyclonal antibody preparation, such as that used in this study, may have dissociation equilibrium constants for the target analyte that span from -8 to 10 -12 M [8]. Therefore, for an antibody-related binding agent with an affinity for its target, a concentration higher than 10 -8 M (i.e., > 1.5 mg L -1 for an intact antibody or > 0.5 mg L -1 for a F ab fragment) should generally be used in a one-site immunometric assay to ensure most of the analyte is complexed during incubation.…”

“…These techniques rely on the selective and strong binding between an antibody and its target to provide assays that can often be used directly, and with minimal pretreatment steps, with complex samples such as serum or plasma [2,3]. In these methods, a column is used that contains one component of an immunoassay, making it possible to combine the binding selectivity and strength of antibodies with the precision, ease of automation, and speed of modern liquid chromatography [2][3][4][5][6][7][8].…”

Enrichment of HDL proteome and phospholipidome from human serum via IMAC/MOAC affinity

“…For lipoprotein and lipopeptide enrichment, several types of affinities have been reported in recent years. Immune‐affinity chromatography and antibodies have been used for the purification of lipoproteins from biological samples (Kawakami, Tanaka, Nakano, Saniabadi, & Numano, ; Zhang et al, ). Later, gel filtration chromatography was employed for the extraction of HDLs from cardiovascular disease plasma samples; however, it lacks selectivity (Gordon, Deng, Lu, & Davidson, ).…”

“…Immune-affinity chromatography and antibodies have been used for the purification of lipoproteins from biological samples (Kawakami, Tanaka, Nakano, Saniabadi, & Numano, 2001;Zhang et al, 2018).…”

Route and Type of Formulation Administered Influences the Absorption and Disposition of Vitamin B12 Levels in Serum

“…Biological and pharmaceutical compounds can encompass peptides, proteins, antigens, antibodies, nucleic acids, lipids, vitamins and minerals, phytochemicals, and other nutraceuticals (e.g., coenzyme Q 10 , glucosamine) as well as cell therapies [1,2,3]. Delivery to target tissues can include oral, pulmonary, subcutaneous, intravenous, transdermal, and nasal routes.…”