Adv Biochem Eng Biotechnol (2020) 174: 93–140
DOI: 10.1007/10_2019_106
© Springer Nature Switzerland AG 2019
Published online: 5 September 2019
Aptamer-Based Affinity Chromatography
for Protein Extraction and Purification
G. Perret and E. Boschetti
Contents
1 Subject Focus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
2 Major Milestones in Affinity Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
3 The Advent of Aptamer Affinity Chromatography Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
3.1 Oligonucleotide Libraries as Ligand Banks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
3.2 Single-Chain Oligonucleotide Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
3.3 Aptamer-Protein Interaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
3.4 The Power of SELEX for Aptameric Ligand Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
4 A Field Yet Largely to Be Explored . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
5 An Update on Aptamer-Based Affinity Chromatography Advancements . . . . . . . . . . . . . . . . . 111
5.1 RNA Versus DNA Aptamer Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
5.2 Aptamer Grafting Methods for Chromatography Exploitations . . . . . . . . . . . . . . . . . . . . . . 112
5.3 A Focus on Major Published Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
5.4 Questions Around Aptamer Release, Hydrolysis, and Sanitization . . . . . . . . . . . . . . . . . . 125
6 Comparison Between Aptamer- and Antibody-Based Affinity Chromatography . . . . . . . . . 126
7 Expectations for Tomorrow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
Abstract Aptamers are oligonucleotide molecules able to recognize very specifically proteins. Among the possible applications, aptamers have been used for affinity
chromatography with effective results and advantages over most advanced protein
separation technologies. This chapter first discusses the context of the affinity
chromatography with aptamer ligands. With the adaptation of SELEX, the chemical
modifications of aptamers to comply with the covalent coupling and the separation
process are then extensively presented. A focus is then made about the most
G. Perret (*)
LFB Biotech, Les Ulis, France
e-mail: perret@netcourrier.com
E. Boschetti
JAM Conseil, Neuilly-sur-Seine, France
DOI: 10.1007/10_2019_106
© Springer Nature Switzerland AG 2019
Published online: 5 September 2019
Aptamer-Based Affinity Chromatography
for Protein Extraction and Purification
G. Perret and E. Boschetti
Contents
1 Subject Focus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
2 Major Milestones in Affinity Chromatography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
3 The Advent of Aptamer Affinity Chromatography Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
3.1 Oligonucleotide Libraries as Ligand Banks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
3.2 Single-Chain Oligonucleotide Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
3.3 Aptamer-Protein Interaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
3.4 The Power of SELEX for Aptameric Ligand Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106
4 A Field Yet Largely to Be Explored . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
5 An Update on Aptamer-Based Affinity Chromatography Advancements . . . . . . . . . . . . . . . . . 111
5.1 RNA Versus DNA Aptamer Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
5.2 Aptamer Grafting Methods for Chromatography Exploitations . . . . . . . . . . . . . . . . . . . . . . 112
5.3 A Focus on Major Published Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
5.4 Questions Around Aptamer Release, Hydrolysis, and Sanitization . . . . . . . . . . . . . . . . . . 125
6 Comparison Between Aptamer- and Antibody-Based Affinity Chromatography . . . . . . . . . 126
7 Expectations for Tomorrow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
Abstract Aptamers are oligonucleotide molecules able to recognize very specifically proteins. Among the possible applications, aptamers have been used for affinity
chromatography with effective results and advantages over most advanced protein
separation technologies. This chapter first discusses the context of the affinity
chromatography with aptamer ligands. With the adaptation of SELEX, the chemical
modifications of aptamers to comply with the covalent coupling and the separation
process are then extensively presented. A focus is then made about the most
G. Perret (*)
LFB Biotech, Les Ulis, France
e-mail: perret@netcourrier.com
E. Boschetti
JAM Conseil, Neuilly-sur-Seine, France