title: Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition.
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).
authors.affiliation: Department of Chemistry, Sogang University.
authors.forename: Sanggil
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Sogang University.
authors.forename: Wooseok
authors.initials: W
authors.lastname: Ko
authors.affiliation: Department of Chemistry, Sogang University.
authors.forename: Hyunji
authors.initials: H
authors.lastname: Park
authors.affiliation: Department of Chemistry, Sogang University; hslee1976@gmail.com.
authors.forename: Hyun Soo
authors.initials: HS
authors.lastname: Lee
mesh_terms: D000480:Alkynes / Q000737:chemistry*; D000596:Amino Acids / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry*; D001386:Azides / Q000737:chemistry*; D003300:Copper; D061565:Cycloaddition Reaction
chemical_list: D000480:Alkynes; D000596:Amino Acids; D000906:Antibodies; D001386:Azides; D003300:Copper