Related Experiment Video
Updated: Sep 25, 2025
11:00
Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
Published on: November 9, 2017
45.4K
A T-cell-dependent antibody response (TDAR) method in BALB/c mice based on a cytometric bead array
Wenhua Zhong1, Penghuan Chang2, Lianfang Gan1
1Hainan Medical University, Research Center for Drug Safety Evaluation of Hainan Province, Haikou, China.
Journal of Immunotoxicology
|April 28, 2022
Summary
A new cytometric bead assay (CBA) offers a more sensitive and precise method for assessing T-cell-dependent antibody responses (TDAR) in mice. This improved assay provides accurate results compared to traditional ELISA methods.
Area of Science:
- Immunology
- Assay Development
Background:
- Current T-cell-dependent antibody responses (TDAR) assays are often semi-quantitative and lack precision.
- Keyhole limpet hemocyanin (KLH) is a common antigen used in TDAR assessments.
- Cytometric bead assays (CBA) offer potential for improved sensitivity, specificity, and convenience.
Purpose of the Study:
- To develop and optimize a novel TDAR assay utilizing CBA technology.
- To enhance the precision, sensitivity, and accuracy of mouse TDAR assessment.
Main Methods:
- Covalently coupled KLH antigen onto cytometric beads using immune microsphere technology.
- Prepared IgM antibody capture spheres for sample pretreatment.
- Optimized key assay parameters including antigen concentration, capture sphere concentration, and detection antibody dilution.
Main Results:
- Achieved excellent linearity (r² = 0.9937) with high precision (within-run: 3.1-4.9%, between-run: 4.4-4.9%).
- Demonstrated a low limit of detection (113.43 ng/ml) within a broad linear range (390.63-50,000 ng/ml).
- Reported a low interference rate (0.04-3.51%).
Conclusions:
- A novel mouse TDAR assay based on CBA has been successfully developed.
- The CBA-based TDAR assay is more sensitive, accurate, and precise than traditional ELISA methods.
- This assay represents a significant advancement for evaluating antibody responses.
