Exploring affinity chromatography in proteomics: A comprehensive review

• Published in: Analytica chimica acta Vol. 1306; p. 342513
• Main Authors: Chamrád, Ivo, Simerský, Radim, Lenobel, René, Novák, Ondřej
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2024
• Subjects: Affinity chromatography; Affinity depletion; Affinity enrichment; Chromatography, Affinity - methods; Humans; Mass spectrometry; Peptide; Protein; Proteins - analysis; Proteins - chemistry; Proteins - isolation & purification; Proteomics; Proteomics - methods; Affinity chromatography; Affinity depletion; Proteomics; Affinity enrichment; Mass spectrometry; Peptide; Protein
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2024.342513

Over the past decades, the proteomics field has undergone rapid growth. Progress in mass spectrometry and bioinformatics, together with separation methods, has brought many innovative approaches to the study of the molecular biology of the cell. The potential of affinity chromatography was recognized immediately after its first application in proteomics, and since that time, it has become one of the cornerstones of many proteomic protocols. Indeed, this chromatographic technique exploiting the specific binding between two molecules has been employed for numerous purposes, from selective removal of interfering (over)abundant proteins or enrichment of scarce biomarkers in complex biological samples to mapping the post-translational modifications and protein interactions with other proteins, nucleic acids or biologically active small molecules. This review presents a comprehensive survey of this versatile analytical tool in current proteomics. To navigate the reader, the haphazard space of affinity separations is classified according to the experiment's aims and the separated molecule's nature. Different types of available ligands and experimental strategies are discussed in further detail for each of the mentioned procedures. [Display omitted] •Affinity chromatography separates molecules based on their specific interactions.•In proteomics, affinity separation aids protein and peptide depletion or enrichment.•Three main areas of proteomic affinity chromatography are explored in detail.•Recent developments in affinity ligands and approaches in proteomics are discussed.•Applications and future potential of affinity proteomics are highlighted.