Affinity chromatography: A review of trends and developments over the past 50 years

Abstract

The field of affinity chromatography, which employs a biologically-related agent as the stationary phase, has seen significant growth since the modern era of this method began in 1968. This review examines the major developments and trends that have occurred in this technique over the past five decades. The basic principles and history of this area are first discussed. This is followed by an overview of the various supports, immobilization strategies, and types of binding agents that have been used in this field. The general types of applications and fields of use that have appeared for affinity chromatography are also considered. A survey of the literature is used to identify major trends in these topics and important areas of use for affinity chromatography in the separation, analysis, or characterization of chemicals and biochemicals.

Keywords: Affinity chromatography; Affinity ligands; Affinity supports; Applications; Immobilization methods.

Figure 1.

The (a) sample application/washing steps and (b) examples of elution methods that are used the on/off mode of affinity chromatography. The three types of elution shown in (b) are non-specific elution, isocratic elution, and biospecific elution. Methods for biospecific elution can be further divided into normal-role elution, in which a competing agent binds to the target, and reversed-role elution, in which the competing agent binds to the immobilized affinity ligand.

Figure 2.

Number of publications including the phrase “affinity chromatography” and that appeared between 1968 and 2019. These data were obtained through a search that was conducted in May 2019 using SciFinder. Similar trends, but with approximately twice the number of papers, were seen when a search was conducted for papers that included the concept of “affinity chromatography” (see Supplementary Material).

Figure 3.

Use of various supports in affinity chromatography, based on a search of papers that have employed this method. This search was conducted in May 2020 using SciFinder and examining papers that appeared between 1968 and 2019 with the phrase “affinity chromatography” used in close association with a particular support. Similar trends to those shown in this plot were seen when the given support and phrase “affinity chromatography” appeared in the same paper but were not necessarily closely associated with each other (see Supplementary Material). The group listed here as “other polymers” included polymethacrylate (used in 0.3% of the papers), polysulfone (0.3%), and polyamide (0.4%). The general structures or chemical formulas for agarose, cellulose, silica, and titania are included for reference.

Figure 4.

Use of various support formats in affinity chromatography, based on a search of papers that have employed this method. This search was conducted in May 2020 using SciFinder and examining papers that appeared between 1968 and 2019 with the phrase “affinity chromatography” used in close association with a particular support format. Similar trends to those shown in this plot were seen when the given format for the support and phrase “affinity chromatography” appeared in the same paper but were not necessarily closely associated with each other (see Supplementary Material).

Figure 5.

Examples of general strategies that may be used to immobilize or place a binding agent, or affinity ligand, within a support for use as a stationary phase in affinity chromatography.

Figure 6.

Extent of use of representative biological and non-biological binding agents in affinity chromatography, based on a search of papers that have employed these agents. This search was conducted in June 2020 using SciFinder and examining papers that appeared between 1968 and 2020 with the phrase “affinity chromatography” or “chromatography” used in close association with the given type of binding agent. The plot on the left includes the results from 10,521 papers, and the plot of the right is based on 7,516 papers. Abbreviations: Ig, immunoglobulin; IMAC, immobilized metal-ion affinity chromatography; MIPs, molecularly imprinted polymers; IAM, immobilized artificial membrane.

Figure 7.

Fields in which affinity chromatography is often used in chemical or biochemical separations, isolation, analysis, or characterization. These results are based on a literature search made using SciFinder during January 2019 for areas with papers in the listed fields that were linked to the term “affinity chromatography”. The percentages shown in this graph are based on a total of 46,335 papers that were obtained during this search.