Related Experiment Video
Updated: Jan 16, 2026
09:04
Measurement of T Cell Alloreactivity Using Imaging Flow Cytometry
Published on: April 19, 2017
13.9K
Development of a novel assay for antigen presentation measurement
Mei Li1, Falak Harshit Sharma1, Yi-Ling Chen2
1Department of Biological Sciences, Texas Tech University, Lubbock, TX, 79409, USA.
Scientific Reports
|September 26, 2025
Summary
A new Click chemistry method offers a faster, cost-effective way to measure antigen presentation by immune cells. This technique improves antigen stability and enables identification of tumor peptides for immunotherapy.
Area of Science:
- Immunology
- Biochemistry
- Chemical Biology
Background:
- Accurate measurement of antigen presentation is crucial for understanding immune responses to infections and tumors.
- Current methods for assessing antigen presentation are often cumbersome, time-consuming, and limited by reliance on known sequences and antibodies, leading to unstable results.
- Existing techniques can result in antigen loss during processing and editing, compromising the reliability of immune response assessments.
Purpose of the Study:
- To develop a novel, cost-effective, and efficient method for examining antigen presentation using Click chemistry.
- To overcome the limitations of existing antigen presentation assays, including speed, cost, stability, and detection consistency.
- To validate the utility of this new method in various antigen-presenting cells (APCs) and antigen types, including potential tumor antigens.
Main Methods:
- Utilized Click chemistry, a bioorthogonal reaction between azides and alkynes/cyclooctenes, for antigen labeling and detection.
- Pre-labeled antigens (BSA, bacteria, tumor antigens) with azides or alkynes for uptake by APCs (RAW264.7, DC2.4, BMDCs).
- Detected antigen presentation using fluorophore-conjugated probes and validated results with fluorescent plate reader, flow cytometry, and ELISA.
Main Results:
- Demonstrated efficient and stable antigen presentation on the surface of all tested APCs.
- Showed enhanced stability of Click chemistry-labeled antigens within APC phagolysosomes.
- Found that azidohomoalanine (AHA)-labeled antigens exhibited superior MHC class II presentation compared to N-terminally labeled antigens, preserving natural antigen editing.
Conclusions:
- The novel Click chemistry-based antigen presentation assay is faster, more cost-effective, and provides more stable and reliable detection than existing methods.
- This technique preserves natural antigen editing processes, facilitating the selection of high-affinity antigens for MHC presentation.
- When combined with mass spectrometry, this assay can identify stably presented tumor peptides, offering potential therapeutic targets for immunotherapy development.
