Efficient and Site-specific Antibody Labeling by Strain-promoted Azide-alkyne Cycloaddition.
J Vis Exp
; (118)2016 12 23.
Article
en En
| MEDLINE
| ID: mdl-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).
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Azidas
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Alquinos
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Reacción de Cicloadición
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Anticuerpos
Idioma:
En
Año:
2016
Tipo del documento:
Article