FACS-Positive Hybridoma Screening

Flow Cytometry

Flow cytometry (FACS) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. Recent years, FACS has been quickly gaining ground over classical immunoassays for the fast detection of disease markers in liquid samples such as fluids, blood, and plasma.

Antibodies used in FACS are typically used to detect low abundance markers or determine the efficiency of specific treatments, substantially aiding the efforts of developing targeted or personalized therapeutic approaches. FACS can be performed with fixed or unfixed samples and they typically employ fluorescence-labeled antibodies in multiplexing conditions. Due to its enhanced sensitivity, off-target binding can become problematic in FACS. For this reason, validation should foresee potential issues caused by different sample types, cell fixation protocols, and multiplexing conditions.

Functional Application Validation

Creative Biolabs helps you achieve your unique project goals by choosing the most robust and flexible solution for diagnostic antibody development. All diagnostic antibodies generated in our hybridoma platform are optimized and validated in the application of your choice considering assay format and conditions, sample composition, and processing protocol. In this way, we ensure all antibodies keep the highest level of performance. Figure 1 shows our hybridoma development service content. Determination of the best clones for ELISA will be done first. Subsequently, FACS assays will be done directly in the target application. Selected clones are subcloned for further antibody production.

Fig.1 Hybridoma development services workflow.

When validating antibodies for FACS, Creative Biolabs optimizes staining protocol, including fixation, permeabilization, and washing. We include relevant controls routinely in our experiment to determine non-specific binding of an antibody. FACS is performed to identify if an antibody has binding activity to the target protein expressed in transfected cells and determine the optimal binding concentration of the antibody. Optimal binding concentration is defined as the lowest concentration where the antibody binds in a saturating manner as indicated by a cell population shift depicted in a FACS histogram overlay. This shift is measured by comparing antibody binding between transfected and untransfected HEK cells.

The unique advantages of using hybridoma generation for diagnostic applications

A robust and mature approach with a proven track record for the cost-effective generation of high-quality antibodies.
Higher stability, makes them ideal when testing the presence of specific markers even in harsh and complex environments.
Highly efficient in vivo affinity maturation.

Creative Biolabs provides fit-for-purpose hybridoma development services for worldwide clients. If you are looking for custom antibody for FACS use, please contact us for more details about our custom services.

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

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