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Bioorthogonal Chemistry: Enzyme Immune and Protein Capture for Enhanced LC-MS Bioanalysis.

Xiaotong Wang^1,2,3, Duanmin Hu³, Perry G Wang⁴

¹Department of Hepatology and Gastroenterology, The Affiliated Infectious Hospital of Soochow University, Suzhou 215004, China.

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|October 29, 2024

Summary

Chemoenzymatic immunocapture and protein capture (IPC) enhances mass spectrometry bioanalysis. This innovative method improves sensitivity and specificity for analyzing diverse biomolecules in drug discovery and diagnostics.

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Glycopeptide Capture for Cell Surface Proteomics

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Area of Science:

Analytical Chemistry
Biochemistry
Biotechnology

Background:

Immunocapture liquid chromatography-mass spectrometry (IC-LC-MS) is vital in drug discovery and diagnostics.
Traditional methods face limitations in sensitivity, specificity, and MS compatibility.
Chemoenzymatic immunocapture and protein capture (IPC) offers enhanced specificity and versatility.

Purpose of the Study:

To review foundational principles and modification strategies of chemoenzymatic IPC.
To highlight advancements and expanded applications of chemoenzymatic IPC.
To introduce a novel solid-phase chemoenzymatic IPC assay (SCEIA).

Main Methods:

Exploration of chemoenzymatic IPC principles.
Review of modification strategies: bioorthogonal click-chemistry, enzymatic-tagging, HaloTag/CLIP-tag.
Development and proposal of a novel solid-phase chemoenzymatic IPC assay (SCEIA).

Main Results:

Chemoenzymatic IPC significantly improves analytical performance for various biomolecules (small molecules, peptides, RNAs, proteins).
Expanded applicability in drug discovery, clinical diagnostics, and environmental analysis.
The proposed SCEIA demonstrates efficient IPC and target analyte release.

Conclusions:

Chemoenzymatic IPC overcomes limitations of traditional methods, offering robust, sensitive, and versatile bioanalysis.
This technique represents a paradigm shift in IC-LC-MS bioanalysis.
Future advancements in bioanalysis are enabled by chemoenzymatic IPC.