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Microbead-Based Platform for Multiplex Detection of DNA and Protein
Xiangjun Liu1,2, Tao Bing1, Dihua Shangguan1,2
1Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences , Beijing 100190, China.
ACS Applied Materials & Interfaces
|March 2, 2017
Summary
We developed a microbead detection platform for sensitive, simultaneous biomolecule analysis. This universal strategy offers high selectivity for DNA and protein targets, even in human serum.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Molecular Diagnostics
Background:
- Multiplexed detection of biomolecules is crucial for diagnostics and research.
- Existing methods often face limitations in sensitivity, selectivity, or ease of use.
- A universal platform for simultaneous analysis of diverse targets is highly desirable.
Purpose of the Study:
- To develop a novel, simple, and universal microbead-based platform for simultaneous biomolecule detection.
- To demonstrate the platform's high sensitivity and selectivity for DNA and protein targets.
- To validate the platform's applicability in complex biological samples like human serum.
Main Methods:
- Utilized streptavidin-coated polystyrene microbeads with dye- and biotin-labeled ssDNA or aptamer probes.
- Employed quencher-labeled complementary sequences; target binding triggers quencher dissociation and fluorescence reactivation.
- Monitored fluorescence variations using flow cytometry for sensitive detection.
Main Results:
- Achieved a low detection limit of 0.20 nM for target DNA with excellent selectivity against mismatched sequences.
- Demonstrated high sensitivity and selectivity for protein detection, with a 8.56 nM detection limit for lysozyme.
- Successfully performed simultaneous detection of multiple targets and validated the assay in human serum samples.
Conclusions:
- The developed microbead platform offers a simple, universal, and highly sensitive strategy for multiplexed biomolecule detection.
- The platform exhibits robust performance for both nucleic acid and protein targets, adaptable through probe variation.
- This technology holds significant potential for various diagnostic and research applications, including analysis in complex biological matrices.
