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Veterinary Therapeutic Antibody Single B Cell Platform
Veterinary therapeutic antibodies are attracting extensive interest from researchers in many fields, and more mature monoclonal antibody preparation methods have emerged. BioVenic has established the veterinary therapeutic antibody single B cell platform, which directly isolates B cells at the single cell level, and then analyzes and screens them, to accurately and efficiently harvest the B cells secreting the target veterinary therapeutic antibody molecules.
Fig.1 Schematic diagram of single B-cell sorting based on FACS and droplet microfluidics for monoclonal antibody production.1,2
Cell Sorting
BioVenic offers high-quality animal models tailored to desired target therapeutic species. We also develop immunization protocols that match the requirements for immunized animals and antigens. Peripheral blood is collected from the immunized animals, and the rough isolation and enrichment of B cells are completed. BioVenic provides various methods for the selection of single B cells. Each method is tailored to ensure the precise isolation of individual B cells based on specific markers or functional characteristics, enabling the generation of monoclonal antibodies with desired properties for veterinary therapeutic applications.
- Fluorescence-Activated Cell Sorting (FACS)
This is the most popular method for single B cell selection within BioVenic, which is based on advanced flow cytometers. If you require antigen-specific single B cell selection, we incorporate a fluorescently labeled target antigen to bind with membrane antibodies on the target B cells, allowing for the selection of the B cells you need.
It uses a laser beam to excite fluorochromes coupled with antibodies bound to the cell's surface. The emitted fluorescent signals are automatically analyzed or sorted based on this technology. While this method may be more complex and has higher equipment and sample requirements, our staff has extensive experience in its operation.
- Magnetic Activated Cell Sorting (MACS)
In the presence of an external magnetic field, cells that are bound to the magnetic beads via the target antigen are attracted and retained within the magnetic field. Cells lacking the surface-specific antibodies that enable binding with the antigen-coated magnetic beads are not magnetic and do not stay within the magnetic field, allowing for cell separation.
This method is based on the principle that cell surface-specific antibodies can bind to the target antigen linked to magnetic beads. Magnetic bead matrix materials provided by BioVenic are biodegradable, sterile, low in endotoxins, and offer a rapid processing time. High-purity, unlabeled B cells can be obtained within 15 minutes.
- Droplet Microfluidics
Droplet microfluidics is a microfluidic technology that BioVenic utilizes to enable high-throughput single-cell sorting and analysis. The service helps prevent cross-cell interference, enhancing experimental accuracy and assisting in rapidly screening and analyzing potential antibody producers in single B cells.
It involves encapsulating individual B cells within tiny water droplets, where each droplet serves as a microreactor. Different antigens can be introduced into each droplet to induce antibody production by B cells. This approach facilitates the high-throughput screening of various single B cells to identify those producing specific antibodies.
Amplification and Cloning of Antibody Genes
Amplification of unknown antibody gene sequences from single B cells involves the use of suitable primers for nested or semi-nested RT-PCR. The design of primers is crucial for amplifying the segments necessary to obtain the complete antibody sequence. BioVenic provides antibody gene amplification and cloning services, including primer design services, and offers primer sequences tailored to different animal species and various antibody types, meeting specific needs.
Antibody Expression
BioVenic has established various veterinary therapeutic antibody expression systems, including prokaryotic and eukaryotic systems. These diverse systems allow for the tailored expression of antibodies to meet specific requirements, such as proper folding, glycosylation, and post-translational modifications, which are critical for the antibody's functionality and therapeutic efficacy. Our team ensures optimal expression conditions for each system, maximizing the yield and purity of the expressed antibodies.
| Prokaryotic Expression System | Simple and commonly used system, typically in Escherichia coli. Antibody gene expression in this system is often in the form of antigen-binding fragments (Fabs). |
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| Eukaryotic Expression System | Including yeast expression, insect expression, and mammalian cell expression. The mammalian system is highly favored for projects, inducing high-efficiency protein expression, ensuring proper protein folding, allowing for complex glycosylation modifications, and is free from endotoxin contamination. The antibodies obtained using this system are expressed in their complete IgG form. |
Antibody Purification
BioVenic utilizes advanced purification techniques, including affinity chromatography, which leverages the specific binding properties of antibodies to an immobilized ligand, and ion-exchange chromatography, which separates antibodies based on their charge differences, to ensure the highest purity levels of expressed antibodies.
| Affinity Chromatography | Ion-Exchange Chromatography |
Antibody Validation
BioVenic offers comprehensive antibody validation services to confirm the specificity, affinity, and functional activity of the antibodies. These meticulous validation processes are designed to guarantee that the antibodies meet the stringent requirements for therapeutic applications, ensuring their efficacy and safety for veterinary use.
| Initial Validation |
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| Biological Activity Validation |
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Why Choose Us?
BioVenic provides you with immunization and test protocols for multi-species animals and helps the veterinary monoclonal antibody development.
A variety of cell sorting methods and antibody expression systems are available for you to choose from.
Provide a package of services from antigen preparation to monoclonal antibody production to simplify the development process.
Each B cell can only produce one antibody to a specific antigen, monoclonal antibodies isolated from single B cells have very high antigen specificity. BioVenic has established the veterinary therapeutic antibody single B cell platform for use in various animal species. If you need this service, please don't hesitate to contact us!
Reference
- Fei, Chenjie, et al. "Potential applications of fluorescence-activated cell sorting (facs) and droplet-based microfluidics in promoting the discovery of specific antibodies for characterizations of fish immune cells." Frontiers in Immunology 12 (2021): 771231.
- Image retrieved from Figure 1 "A schematic overview of recombinant mAbs production using FACS and droplet-based microfluidics. B lymphocytes (e.g. splenocytes and plasmablasts) are collected from immunized mice and then individually sorted via FACS or droplet-based microfluidics for antigen-specific lymphocytes.", Fei, Chenjie, et al., 2021, used under [CC BY 4.0], the image title was changed to "Schematic diagram of single B-cell sorting based on FACS and droplet microfluidics for monoclonal antibody production."