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Hybridoma Generation for Therapeutic Antibody Discovery Services

Murine Hybridoma Technology, Generating Mouse Hybridoma Cells for Monoclonal Antibody Production

Catalog No. Product Name Size List Price (US$) Quantity
BS023B Rat Hybridoma Generation for Therapeutic Antibody Discovery 1 package Request
BS023C Hamster Hybridoma Generation for Therapeutic Antibody Discovery 1 package Request
BS023A Mouse Hybridoma Generation for Therapeutic Antibody Discovery 1 package Request

Hybridoma technology is widely used for generating murine monoclonal antibodies for research reagent, diagnostic and therapeutic purposes. Syd Labs provides the most comprehensive murine hybridoma generation for therapeutic antibody services with combination of the following factors:

Genetic background: white, black, immunodeficiency, knock-out, and humanized mice; rat; hamster;

Antigens: proteins, peptides, DNA, cells expressing certain proteins;

Hybridoma fusion: electro-fusion, PEG-fusion;

Antibody screening: ELISA, flow cytometry.

BS023A: a standard program for mouse hybridoma generation for therapeutic antibody discovery.

Phase I: Immunization and boosts of 5 Balb/c mice, 3 following the repetitive immunizations multiple sites or RIMMS protocol and 2 following the standard immunization protocol, and ELISA screening of mouse polyclonal sera.
Phase II: Electrofusion of splenocytes to mouse myeloma cells and ELISA screening to isolate antigen-positive hybridoma cells from an initial count of about 30000 fusion clones. Supernatant (conditioned media) from the final ELISA-positive clone population pools will be sent to clients for evaluation of specific applications.
Phase III: Subcloning of the ELISA-positive clone population pools by limited dilution to isolate individual hybridoma clones of interest, and expansion of the individual hybridoma clones.

You provide: 10 mg of peptide (purity > 85%, length > 12 amino acids) or 5 mg of soluble protein (concentration > 0.4 mg/ml, purity > 90%, molecular weight > 20 kDa) for immunization and screening.

* About 80 ul supernatant from 50-100 ELISA positive cells for evaluation;
* 10 positive hybridoma cell lines;
* Remaining antigens;
* ELISA results.

Questions and Answers:

Question: Do you have a lab facility in US for your murine hybridoma generation services?
Answer: Syd Labs has always tried its best to save clients' cost and provide convenience to clients. Syd Labs has provided various polyclonal and monoclonal antibody production services from facilities in China with the lowest prices in the past decade. Syd Labs added labs for monoclonal antibody services in California and Massachusetts to serve local clients in US, including San Francisco, San Diego, Boston, and New York.

Question: What is your advantage of hybridoma generation services in US?
Answer: Many US clients prefer to work with CRO companies (Contract Research Organization) in US for various reasons. For example, clients can do animal immunization by themselves or university core facilities, and send the spleen to Syd Labs for hybridoma fusion and later hybridoma subcloning and expansion. Polyclones from hybridoma fusion should not be frozen before subcloning because it is easy to lose the monoclone to pursue during the process. The cells must keep growing during screening and activity confirmation. If a client prefer to screen antibodies by themselves with their own functional assays such as flow cytometry, the turnaround time from supernatant shipping to screening results back has to be no longer than 4 days. For such a collaboration process of custom monoclonal antibody development, it is better to have a domestic CRO like Syd Labs.

Question: Can you do DNA immunization for hybridoma generation?
Answer: Yes, Syd Labs does hybridoma generation for monoclonal antibody production with immunization from DNA. Please request quote for DNA immunization. Syd Labs can also use peptides, small molecular compounds, proteins, and cells expressing a protein as the antigens. One needs to do KLH or BSA conjugation if the molecular weight of the antigen is smaller than 20 kDa. During mouse immunization for monoclonal antibody development, Syd Labs may make a combination of various immunogens such as immunization with DNA or cells and boost with proteins.

Question: Can you do RIMMS for mouse immunization?
Answer: Yes, Syd Labs uses Repetitive IMmunizations Multiple Sites (RIMMS) as the alternative immunization protocol to the standard immunization protocol of mouse monoclonal antibody production. Please request quote for RIMMS immunization. Normally the timeline of RIMMS immunization is shorter than 30 days.

Question: What are the most important parts in custom monoclonal antibody development?
Answer: No matter if clients develop monoclonal antibodies in house or by CRO, one must have two things: one is a reliable high-throughput assay to evaluate antibody candidates, and the other great results of animal immunization. Depending on clients' antibody application, one needs to develop:  functional assays for therapeutic application, immunoassay with real samples for clinical diagnostics application, and tissue microarray for reagent development.

Syd Labs also provides other monoclonal antibody development services:

Rat monoclonal antibody development services (rat hybridoma generation)
Camelid monoclonal antibody development services

After great hybridoma clones have been developed, if clients want to file patents, protect the antibodies using the digital format, or develop antibody drugs, clients can ask us to clone and sequence the hybridoma antibody variable regions (VH and VL), do transient expression of chimeric antibodies (scFv or complete IgG) in CHO or HEK293 cells, or generate stable cell line expressing antibodies.

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