SpeedeeTM Monoclonal Antibody Production

Seattle Genova offers custom monoclonal antibody development services using hybridoma technology. Our facility generates high-affinity custom antibodies with industry-leading titer guarantees and comprehensive antibody production packages that simplify the ordering process and present cost-effective methods to isolate epitope-specific antibodies.

Our monoclonal antibody production services include - immunization, cell fusion, cloning, characterization, and production of your monoclonal antibody in vivo in BALB/c or immunocompromized (e.g., SCID, Nu/Nu, RAG-/-) mice, or in vitro in flasks or bioreactors. You retain all rights to any clones that are produced. Once the clones are developed, we can scale them up as tissue culture supernatant.

Highlights

• Monoclonal antibodies—optimized and efficient protocols

• Guaranteed titer—we will continue to boost until a quantifiable minimum titer is achieved

• Full-service integration—combine these protocols with our complete set of services for initial antigen preparation and subsequent antibody production and purification

Standard development process

Phase 1: Antigen preparation

You can provide purified protein (SDS-PAGE purity >90%) or peptide/small molecule antigens that are ready for immunization. 

Or we can provide services for protein production, peptide synthesis, and peptide/small molecule conjugation to carrier protein for use as the immunogen.

Phase 2: Immunization and ELISA titration

One primary injection and two booster injections are performed over a 6-week period. A titration ELISA is performed before each booster. ELISA results and crude antisera are sent to the customer for evaluation. 

Phase 3: Fusion and screening for positive supernatants

The animals are boosted twice more, then their spleen cells are harvested for hybridoma fusion. An ELISA assay is used to screen the positive supernatants, 1 to 2 mL of which are sent to the customer for evaluation. 

Phase 4: Subcloning before production

Parental cell lines can be subcloned through limiting-dilution cloning to obtain daughter cell lines. Multi-cycle complete clonality cloning involves performing multiple rounds of limiting-dilution subcloning until all wells are growing at the same rate, are secreting antibody at the same concentration, and have the same confirmed isotype. 

Phase 5: Production and purification

Hybridoma cells are cultured in shaking flasks to yield antibody-containing supernatants. Protein A or Protein G resin affinity purification is performed after cell culture. Low-endotoxin and serum-free culturing are available on request, as well as high-volume production.