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  • Title: Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition.
    Author: Kim S, Ko W, Park H, Lee HS.
    Journal: J Vis Exp; 2016 Dec 23; (118):. PubMed ID: 28060353.
    Abstract:
    There are currently many chemical tools available to introduce chemical probes into proteins to study their structure and function. A useful method is protein conjugation by genetically introducing an unnatural amino acid containing a bioorthogonal functional group. This report describes a detailed protocol for site-specific antibody conjugation. The protocol includes experimental details for the genetic incorporation of an azide-containing amino acid, and the conjugation reaction by strain-promoted azide-alkyne cycloaddition (SPAAC). This strain-promoted reaction proceeds by simple mixing of the reacting molecules at physiological pH and temperature, and does not require additional reagents such as copper(I) ions and copper-chelating ligands. Therefore, this method would be useful for general protein conjugation and development of antibody drug conjugates (ADCs).
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