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Standardization of a Cytometric Bead Assay Based on Egg-Yolk Antibodies
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Direct Screening for Cytometric Bead Assays for Adenosine Triphosphate
Hao Qu1,2, Lu Wang1, Jian Liu1
1School of Food and Biological Engineering , Hefei University of Technology , Hefei , Anhui 230009 , China.
ACS Sensors
|August 8, 2018
Summary
We developed a novel screening method to rapidly discover aptamer-based cytometric bead assays (AB-CBAs). This technique efficiently identifies functional aptamer particles for diverse analytes, overcoming limitations of traditional methods.
Area of Science:
- Biotechnology
- Molecular Biology
- Analytical Chemistry
Background:
- Cytometric bead assays offer advantages but face challenges with molecular recognition element immobilization.
- Immobilization can compromise the function of antibodies and aptamers, limiting assay performance.
Purpose of the Study:
- To develop a rapid and direct screening approach for aptamer-based cytometric bead assays (AB-CBAs).
- To overcome functional failure issues associated with immobilizing molecular recognition elements.
Main Methods:
- Transformed molecular libraries into pools of monoclonal aptamer particles, with each particle displaying identical aptamer sequences.
- Utilized a two-color fluorescent reporter system where aptamer structural changes yield optical readouts.
- Employed fluorescence-activated cell sorting to isolate aptamer particles with significant signal changes upon target binding.
Main Results:
- Successfully isolated AB-CBA aptamer particles with high signaling performance for ATP within three screening rounds.
- Demonstrated the capability to screen up to 10^8 particles per hour.
- Validated the effectiveness of the screening strategy for identifying functional aptamers.
Conclusions:
- The developed screening strategy is powerful for direct and rapid discovery of aptamer-based cytometric bead assays.
- This platform enables the generation of AB-CBAs for a wide range of analytes.
- The method addresses limitations of current immobilization techniques for molecular recognition elements.
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