Hybridoma Enrichment Service

Creative Biolabs is the preferred antibody production partner for our clients who leverage our scientific expertise and flexibility for high-quality hybridoma development. Our tailored facilities and experienced scientists offer hybridoma enrichment and optimization services for global clients.

Hybridoma Enrichment by FACS

Antibody production from established hybridoma cell lines can diminish over time due to a decrease in either the number of cells that secrete antibodies or the variations in the amount of antibody secreted. In many cases, antibody production can be improved by using different techniques which include the identification and isolation of FACS enriched population of hybridomas producing the uppermost amount of antibody in the culture. Multiple rounds of FACS enrichment will be carried out and the top antibody-producing cells are expanded and frozen.

Available Approaches

In this strategy, scientists conjugate ﬂuorescent particles with the antigen and let them bind to hybridomas that present the specific antibody on their surface. In this way, only antigen-specific hybridomas bind the particles and become ﬂuorescently labeled, allowing cytometric sorting. This approach relies on strong antibody display on the cell surface, as the ﬂuorescent signal is proportional to the number of labeled antigen molecules that bind to it. One can select specific hybridomas and even for more advanced properties such as isotype, and specificity versus universality using multiple antigens.

Fig.1 An illustration of the hybridoma sorting method 1.¹

In order to detect antigen specificity, hybridoma cells are incubated with a hapten-horseradish peroxidase conjugate (namely hapten-HRP), which is subsequently incubated with a fluorophore-labeled polyclonal anti-peroxidase antibody. To characterize the expression of membrane-bound immunoglobulin G (IgG), a fluorophore-labeled anti-mouse IgG antibody is used. Hundreds of labeled hybridoma cells producing mAbs specific for a hapten are rapidly isolated and deposited from a fusion mixture as single-cell clones via FACS. ELISA measurements of the supernatants of the sorted hybridoma clones are further conducted.

Fig.2 An illustration of the hybridoma sorting method 2.

Single-cell Hybridoma Screening using Droplet-based Microfluidics

Monoclonal antibodies (mAbs) can specifically bind or even inhibit drug targets and have hence become the fastest growing class of human therapeutics. Although they can be screened for binding affinities at very high throughput using different systems, screening for functional properties is much more challenging. Generally, these screens require the generation of immortalized hybridoma cells, as well as clonal expansion in microtiter plates over several weeks, and the number of clones that can be assayed is typically no more than a few thousand. Nowadays, droplet-based microfluidics has been proposed to screen single-cell hybridoma. Individual cells are encapsulated into aqueous microdroplets and assayed directly for the release of antibodies inhibiting a drug target based on fluorescence.

Besides the mentioned hybridoma enrichment services, Creative Biolabs also offers multiple hybridoma rescue services and comprehensive mAb preparation services based on our diverse hybridoma platform. If you are interested in our services, please don't hesitate to contact us for more information.

Reference

Sittner, Assa, et al. "Improved production of monoclonal antibodies against the LcrV antigen of Yersinia pestis using FACS-aided hybridoma selection." Journal of Biological Methods 5.4 (2018).

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

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