FACS Positive Hybridoma Clone Screening Introduction

The common limited dilution method cannot meet the needs of rapidly evolving biological research. As a result, new techniques have been developed to screen hybridoma cells with high quality and speed, and flow cytometry (FACS) is one of them. We offer a comprehensive hybridoma platform to fully support our customers' research needs.

Principles of Hybridoma Screening by Flow Cytometry

Here, we briefly describe the principle of FACS screening of hybridoma cells. First, the cells are dispersed to ensure the cell samples are free of cell clumps. Then, a fluorescent signal is labeled on the cell surface or within the cells. When a single cell passes through the glass capillary individually under pressure, the instrument can automatically detect the fluorescence signal intensity of the individual cell. The fluorescence signal intensity of the cell reflects the corresponding antibody expression level of this cell for screening purposes. Finally, cells with high fluorescence signal intensity are collected, and then monoclonal clones with a high level of expression are obtained by methods such as limited dilution, or single hybridoma cells with a high level of expression can also be collected using a single cell collector.

Advantages of FACS-Positive Hybridoma Screening

The currently commonly used method for the second screening of hybridomas is the limited dilution method. Although this method is indeed able to complete the second screening of hybridomas, it has many drawbacks, such as manual operation, tedious and time-consuming work prone to errors, the inability to guarantee monoclonality, and the need to subclone 3-4 times. In addition, since limited dilution is done before ELISA detection, a minimum of one ELISA identification is done for all clones. In contrast, the second screening of hybridoma cells using FACS greatly simplifies the entire screening process.

FACS can achieve automated screening, which ensures the correctness and accuracy of experimental results.
High efficiency. By using FACS, it is possible to screen for more clones in less time.
Automated screening of hybridoma cells greatly saves human resources and valuable research time, allowing for more projects to be run simultaneously.

Limitations of FACS-Positive Hybridoma Screening

Although FACS has many advantages in screening hybridoma cells, we must still look at the technique objectively. Therefore, we summarize the disadvantages of FACS for screening hybridoma cells so that you can choose the right screening method rationally according to your experimental protocol.

Since fluorescent labeling must be used on the cell surface or inside, it is generally used mainly for intracellularly expressed proteins or membrane-expressed proteins. For secreted-expressing hybridoma cells, its use has limitations.
The specific detection is the fluorescence intensity of a single cell at a certain time point. Therefore, the assay results are easily influenced by the cell cycle. Moreover, the expression level of a single cell does not reflect the expression level of the cell population at the later production stage.
Flow screening technology only separates all cells with a strong fluorescence signal, and the number of cells that need to be rescreened later is still large, and the downstream workload is still relatively large.
As the pressure operation is used on individual cells, the shear force will reduce the cell survival rate. Therefore, the low survival rate also becomes a major drawback of this technique.
Flow screening requires adequate cleaning and disinfection of the pipeline system to ensure that cells are not contaminated. Therefore, the risk of contamination is high.

Creative Biolabs would like to share more successful experiences with FACS-positive hybridoma screening to help your research run smoothly.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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