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Flow cytometric study of hybridoma cell culture: correlation between cell surface fluorescence and IgG production
1Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis 55455.
Insights
Flow cytometry quantifies IgG on hybridoma cells using FITC-conjugated antibodies. This method differentiates antibody-producing cells and correlates fluorescence intensity with antibody production rates.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Hybridoma technology is crucial for monoclonal antibody production.
- Accurate methods are needed to identify and quantify antibody-producing hybridoma cells.
- Cell surface marker analysis can offer insights into cellular function.
Purpose of the Study:
- To develop and validate a flow cytometry-based method for assessing hybridoma cell antibody production.
- To correlate cell surface fluorescence intensity with specific antibody production rates.
- To differentiate between antibody-producing and non-producing hybridoma cells.
Main Methods:
- Hybridoma cells were stained with FITC-conjugated anti-mouse IgG F(ab')2.
- Flow cytometry was employed to measure IgG fluorescence on the cell surface.
- Light scattering techniques were used to exclude dead cells from analysis.
Main Results:
- Surface fluorescence intensity successfully differentiated between antibody-producing and non-producing hybridoma cells.
- A linear correlation was established between mean surface fluorescence intensity and specific antibody production rate.
- The method demonstrated sensitivity in quantifying antibody production at the single-cell level.
Conclusions:
- Flow cytometry provides a robust and quantitative method for evaluating hybridoma cell antibody production.
- This technique facilitates efficient screening and selection of high-producing hybridoma clones.
- The findings support the use of flow cytometry in optimizing monoclonal antibody development processes.
Abstract:
Flow cytometry was applied to measure the fluorescence of IgG molecules on hybridoma cell surface stained with FITC-conjugated anti-mouse IgG F(ab')2. Using light scattering to exclude the dead cell population from the analysis, the surface fluorescence intensity allows for the differentiation between producing and nonproducing cells. A linear correlation has been found between the intensity of mean surface fluorescence of the cell population and the specific antibody production rate.
