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As antibody developing company, we perform over 500 fusions annually. Our experience is with protein antigens, peptide antigens , haptens, viruses, bacteria and so on. Our services are relatively inexpensive due to well organized lab and use of new technologies, wherever possible. Just a short overview of how we perform fusions:
Antigen
We accept any nontoxic antigens, can perform conjugation of haptens, peptides, provide MAP synthesis, accept soluble and insoluble antigens, antigens on nitrocellulose (as soon as there is testing material as well). Currently we do not perform genetic immunizations with plasmids, but this is coming soon.
Immunizations
We immunize mice (usually at least 4) of single (in that case we suggest Balb/c) or two different strains (Balb/c and F1 -CBC6 mice). Our immunization protocols and all animal experiments are approved by the Ethical Comitee. The permission is valid for USA (NIH requirements) as well. Please inquire the license/protocol No. To evaluate the antigen we perform first bleeding at day 20-21. The sera are collected and analyzed by ELISA (unless other methods were requested, we perform immunoblotting and/or immunohistochemistry as well). The decision about fusion or further immunization (or further vaccination) is made after customers reviews the test bleed report.
Fusions
We perform electrofusion with isolated splenocytes or cells from lymph nodes. On average we use 20% of the splenocytes from a mouse, the remaining splenocytes are frozen and stored for additional fusions, if required. In vitro activation of splenocytes with antigen for increase of the specific antibody producing B-cells is possible but not use routinely. On average we obtain from 600 to 1500 primary clones per 96 well plate. We guarantee certain number of positive clones (depends on the project type). Additional fusions may be performed if required or requested.
Screening and clones selection
Depending on the request of a customer and strength of response we pick all positive clones. The strongest ones are immediately re-cloned, meanwhile the medium from the primary wells is shipped to a customer for testing, if requested. Several clones are delivered for customer, other clones may be stabilized upon request.
Storage
We store all materials (splenocytes, cells, primary clones, secondary clones) for every customer for 6 month for free thus providing the possibility of always come back to the original material, repeat fusions etc.
Other services
We perform in house characterization of antibody, including immunohistochemistry, immunoassays, Western blot, flow cytometry and other services. The antibodies may be purified, freeze-dried for storage, etc.
Pricing
Cost of services is defined by the complexity of the project. Please ask for a quotation. General overview of costs for project/operations is on the Ordering Information Page.
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