In the process of preparing monoclonal antibodies from hybridomas, the screening and enrichment of hybridoma cells are crucial links. It is noteworthy that the production of antibodies by established hybridoma lines gradually decreases over time. Based on this fact, it is very necessary to rescue hybridomas through the enrichment of hybridoma cells.
FACS enrichment relies on fluorescent markers to sort target cells. The principle is that fluorescent markers bind to hybridoma cell surface proteins, and when cells pass through the cell separator, the fluorescent markers are activated. Based on differences in fluorescence intensity and cell size, cells are systematically screened to achieve cloning. FACS simplifies the hybridoma cell enrichment procedure and improves screening efficiency. At the same time, the monoclonality of the hybridoma cells is also guaranteed.
Fig.1 Procedures for FACS analysis of heterotypic CICs.1
Currently, droplet-based microfluidics technology has gradually emerged as an attractive tool for screening single-cell hybridomas. High-throughput screening based on this technology allows for the rapid analysis of thousands of single cells. This technology first pumps a suspension of microbeads into a microfluidic chip to form microdroplets to ensure that individual cells are encapsulated into the microdroplets. Subsequently, the droplets are lysed to release the mRNA hybrid. After reverse transcription, amplification is completed by PCR, and final sequencing is performed. This technology contributes to the accurate enrichment of target hybridoma cells. At the same time, this technology significantly reduces costs and improves enrichment efficiency.
LEAP technology allows the proliferation of antibody-secreting cell lines while eliminating hybridoma cell lines that do not secrete antibodies. The basic principle of this method is that the LEAP instrument can image the cells in a specific culture plate, and through the supporting software, the hybridoma cells secreting the target antibody are screened, while the remaining cells are eliminated by a series of targeted laser pulses.
A: The methods of enrichment mentioned above have their respective advantages and scope of applications. Please discuss with our expert technical support team, and select the appropriate method or customize your project of hybridoma enrichment according to the specific conditions of your hybridoma cell line and your intended results.
A: Final deliverables typically include:
In addition to the hybridoma enrichment services shown above, multiple hybridoma rescue services and comprehensive monoclonal antibody preparation services are also available at Creative Biolabs. Our experienced team is willing to share the key considerations for hybridoma enrichment and empower your research with our advanced hybridoma platforms. If you are interested in our hybridoma enrichment services, please contact us for more details.
All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.