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Novel high-throughput cell-based hybridoma screening methodology using the Celigo Image Cytometer
Haohai Zhang1, Leo Li-Ying Chan2, William Rice2
1Department of Liver Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100730, China; Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, United States.
Journal of Immunological Methods
|April 18, 2017
Summary
This study introduces a novel cell-based image cytometry method for high-throughput hybridoma screening, improving antibody discovery efficiency. The new technique accurately identifies potential antibody clones faster and more reliably than traditional methods.
Area of Science:
- Biotechnology
- Immunology
- Cell Biology
Background:
- Hybridoma screening is essential for antibody discovery but current methods like ELISA and flow cytometry have limitations.
- Existing techniques are time-consuming, costly, and struggle with difficult antigens like membrane proteins.
- There is a need for a more accurate, high-throughput, and efficient screening method.
Purpose of the Study:
- To develop and validate a novel high-throughput screening methodology for hybridoma screening using the Celigo Image Cytometer.
- To overcome the limitations of conventional screening methods, particularly for challenging antigens.
- To improve the speed, accuracy, and efficiency of antibody discovery.
Main Methods:
- Utilized the Celigo Image Cytometer for high-throughput screening of hybridoma supernatants.
- Optimized the method by detecting antibody binding to recombinant CD39 expressed on Chinese hamster ovary (CHO) cells.
- Assessed sensitivity via serial dilution of purified antibody and measured antibody affinities (Kd) to membrane-bound CD39.
Main Results:
- The cell-based image cytometry method achieved high-throughput screening within one day, significantly enhancing efficiency.
- Directly measuring antibody binding on living cells eliminated false positives and negatives.
- The method demonstrated high sensitivity, detecting antibodies at concentrations as low as ~5ng/mL, and enabled Kd determination.
Conclusions:
- The developed image cytometry method offers a sensitive, versatile, and efficient approach for hybridoma screening.
- This cell-based technique significantly advances antibody discovery and screening technologies.
- It provides a robust alternative to conventional methods, especially for membrane-bound or complex antigens.