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Updated: Aug 5, 2026
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Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates
Published on: March 6, 2019
Chemoselective Histidine Bioconjugation Using Thiophosphorodichloridate Reagents
Xinyuan Li1, Fung Kit Tang1, Shang Jia2
1Department of Chemistry, Rutgers University-Newark, Newark, NJ, 07102, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 1, 2026
Summary
We developed a histidine-specific protein labeling method using thiophosphorodichloridate reagents. This technique creates stable linkages and enables further bioconjugation via copper-catalyzed azide-alkyne cycloaddition (CuAAC).
Area of Science:
- Biochemistry
- Chemical Biology
- Protein Chemistry
Background:
- Chemoselective bioconjugation is crucial for protein modification and functional studies.
- Targeted labeling enables precise analysis of protein structure and function.
Purpose of the Study:
- To present a detailed protocol for histidine-specific protein labeling.
- To establish a method for creating stable thiophosphoramidate linkages on proteins.
- To enable subsequent conjugation using click chemistry.
Main Methods:
- Synthesis of thiophosphorodichloridate labeling reagents.
- Histidine-specific protein modification.
- Purification of modified proteins via ion-exchange chromatography.
- Copper-catalyzed azide-alkyne cycloaddition (CuAAC) for secondary conjugation.
Main Results:
- Successful histidine-specific labeling of proteins was achieved.
- Stable thiophosphoramidate linkages were formed.
- The labeled proteins were amenable to further functionalization via CuAAC.
Conclusions:
- This protocol provides a robust method for histidine-specific protein labeling.
- The resulting thiophosphoramidate linkage offers stability for bioconjugation.
- The method facilitates downstream applications in chemical biology and proteomics.
