Affinity Bioorthogonal Chemistry (ABC) Tags for Site‐Selective Conjugation, On‐Resin Protein‐Protein Coupling, and Purification of Protein Conjugates
Year: 2022
Type: article
Abstract: The site-selective functionalization of proteins has broad application in chemical biology, but can be limited when mixtures result from incomplete conversion or the formation of protein containing side products. It is shown here that when proteins are covalently tagged with pyridyl-tetrazines, the nickel-iminodiacetate (Ni-IDA) resins commonly used for His-tags can be directly used for protein affinity purification. These Affinity Bioorthogonal Chemistry (ABC) tags serve a dual role by enabling affinity-based protein purification while maintaining rapid kinetics in bioorthogonal reactions. ABC-tagging works with a range of site-selective bioconjugation methods with proteins tagged at the C-terminus, N-terminus or at internal positions. ABC-tagged proteins can also be purified from complex mixtures including cell lysate. The combination of site-selective conjugation and clean-up with ABC-tagged proteins also allows for facile on-resin reactions to provide protein-protein conjugates.
Institution: University of Delaware
Language: English
Subfield: Organic Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
Awards: G1177608224, Zeiss LSM710 Inverted Confocal Microscope, Spinning Disk Confocal and Single Molecule Localization Microscope, G1705962760, G2418039038, G274834250, Discovery of Chemical Probes and Therapeutic Leads, Phase II, G3115553707, G3356351361, G4444685404, G4624185658, G5681054338, G6168504995, G6481875778, G6772302223, University of Delaware MRSEC - Center for Hybrid, Active, and Responsive Materials (CHARM), Multimodal PerkinElmer IVIS-SpectrumCT imaging system for a preclinical imaging core facility, G8003541617, G8414554082, G8575715201, High Sensitivity NMR Spectrometer with Cryoprobe, G8898404032, G9000006994, Delaware INBRE, Toolkit for Fast, Multipurpose and Inducible Bioorthogonal Chemistry, A Hydrogel-Based Cellular Model of the Human Vocal Fold, Q-Exactive Orbitrap LC-MC/MS System
OpenAlex ID: W4294669256