Hybridoma Enrichment Introduction

In the process of developing monoclonal antibodies based on hybridomas, screening and enrichment of hybridoma cells is inevitable. Particularly, in the case of the gradual decrease of antibodies to established hybridoma cell lines, enrichment of hybridoma cells is one of the most important ways for hybridomas rescue. Creative Biolabs has established a comprehensive hybridoma platform that can provide high-quality and efficient hybridoma enrichment services to help customers obtain more hybridoma antibodies in a shorter period of time.

Hybridoma Enrichment Based on FACS

Fluorescence-activated cell sorting (FACS) allows cells in a sample to be sorted using a fluorescent dye by labeling specific proteins that may be present on certain cells and then removing these cells through a specialized device. This method can greatly simplify the process of hybridoma cell enrichment and reduce the working time, while at the same time ensuring the monoclonality of hybridoma cells and producing a large number of clones.

The principle of FACS enrichment in hybridoma is that a fluorescent secondary antibody is added to a well that can secrete antibodies. The fluorescent secondary antibody can bind to the surface of hybridoma cells, and the fluorescence of the secondary antibody on the cell surface is excited when the cells are passed through the fluorescence-activated cell separator one by one. At this time, the computer system can detect this fluorescence and collect the cells in a hierarchical manner according to the difference between fluorescence intensity and cell size. Finally, cloning is achieved.

Hybridoma Enrichment Based on Droplet-Based Microfluidics

High-throughput screening based on droplet microfluidics is a precaution capable of rapidly analyzing thousands of single cells. The main steps of the method include the following. First, a cell suspension with a microbead suspension with molecular barcodes is pumped into a microfluidic chip to converge and form a laminar flow. Then, it is followed by convergence with the oil phase to form monodisperse microdroplets. Each cell is wrapped in the same microdroplet with one microbead, followed by the completion of RNA hybridization and lysis of the droplet to release the mRNA hybrid and complete reverse transcription. Finally, nucleic acid amplification is completed by PCR and sequencing is performed.

Droplet-based microfluidic technology for high-throughput screening enables rapid separation of microvolume samples, enabling parallel high-throughput analysis of thousands of cells. Typically, 1000 microdroplets can be generated per second, each with a volume of 1 nL. This method can greatly reduce costs and save work time while accurately enriching target hybridoma cells.

Hybridoma Enrichment Based on LEAP

Another method that can enrich hybridoma cells is to directly kill the non-antibody-secreting hybridoma cell lines. Laser-enabled analysis and processing (LEAP) is used by removing the non-antibody-secreting hybridoma cell lines and allowing the antibody-secreting cell lines to proliferate. In this method, hybridoma cells are grown in cell culture plates treated with the capture matrix. Secreted antibodies are captured in the vicinity of the antibody-secreting cells and then quantified by fluorescently labeled antigens. The LEAP instrument can image the cells in the culture plate and identify the hybridoma cells that secrete specific antibodies by using the appropriate software. All other cells will be eliminated by a series of targeted laser pulses.

Scientists at Creative Biolabs would love to share the key points and experiences in hybridoma enrichment and promote the smooth process of your project by leveraging our professional technology and platform.

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

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