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Murine Hybridoma Production

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Creative Biolabs provides rat and mouse monoclonal antibodies construction services. In particular, our proprietary mouse immunization approach allows us to provide mouse monoclonal antibodies within 70 days.

To generate monoclonal antibodies that fit your specific purposes, we tailor our protocols in every major step of antibody production, including antigen preparation [peptide synthesis or protein expression in E.coli, yeast, insect or mammalian cells], animal immunization and hybridoma screening.

Monoclonal Antibody Production: One-Stop Services

Monoclonal Antibody Production: One-Stop Services

Antigen Preparation

• We express and purify your recombinant proteins with an appropriate E. coli, yeast, insect or mammalian cell expression system according to the specific purposes of the final antibodies.
• Our proprietary chimeric protein expression method can produce a recombinant protein immunogen in a soluble form in bacterial cells that retains the specificity and immunogenicity of the original protein, and generate antibodies that bind to the protein surface. This approach highly increases the possibility of getting IHC-positive monoclonal antibodies, and antibodies that recognize the native protein and good for ELISA, IP, IHC, Immuno-fluorescence and Western Blotting.
• We can design membrane mimc protein antigen (MMPA) using MPAT™ platform. It remains structural characteristics such as multi-spanner extracellular domain and it is feasible to express in whole water-soluble form with high purity.
• Alternatively, we perform peptide synthesis and conjugation.

Immunizations

• We immunize animals with customized protocols that involve proper adjuvants, inoculation routes, dosage and timing. We can develop antibodies targeting impure native antigens or rare antigens of a minimum amount by adjusting the immunization strategies.
• Our proprietary Magic™ adjuvant can elicit stronger and faster immune response within only 21 days.
• We use different formats of immunogens for animal immunizations depending on the specific purpose. For antibodies intended for immunohistochemical [or immunocytochemical] staining, in addition to the natural immunogens, e.g. recombinant protein, we may alternatively use denatured antigens in animal immunization and hybridoma screening.
• Our fast track mouse immunization protocol allows us to provide monoclonal antibodies within 70 days.

Development of Hybridoma

• We fuse splenocytes with proper myeloma cells using optimized PEG mediated fusion protocols.
• We screen hybridoma clones with customized protocols according to the final applications of the requested antibodies, which includ but are not limited to immunoprecipitation, immunoblotting, various ELISA and particularly, IHC methods.
• Goal-oriented method to screen IHC positive monoclonal antibodies. We introduce our proprietary IHC-positive hybridoma screening protocols to provide you IHC suited hybridoma clones.
• Our revolutionary platform Omni-HybridomaTM platform ensures that a large number of hybridoma clones can be selected after each cell fusion. It can eliminate the possibility of overgrowth of potentially valuable slow-growing clones by fast-growing clones.
• We deliver your positive hybridoma clones in culture or in cryopreservation as well as the derived monoclonal antibodies.

Ascites Production or in Vitro Antibody Manufacturing

• Inoculation of hybridoma cells into relevant animals to produce ascites. We normally use immuno-sufficient mice to produce ascites. Immuno-deficient mice are available for ascites production for non-murine hybridoma cell lines or murine cell lines of distinct genetic backgrounds. Optimized methods are used to generate tumors and accumulate ascitic fluid. Using nude or SCID mice in ascites production will eliminate the contamination from endogenous mouse IgG.
• Cell culture in protein-free medium. We use stationary flasks or 5-50liter bioreactors to manufacture monoclonal antibodies. We have an antibody fermentation capacity of over 300L.




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