Monoclonal Antibody Development The standard service includes: 1. Discussion of the project (screening system, selection criteria, etc.) and planning the work. 2. Immunization, sera collection and ELISA identification 3. Cell Fusion screening and selection of the best cultures for cloning (up to 5) 4. Cloning, subcloning (about 3 rounds of limit dilutions) and screening 5. Cell storage (cryopreservation) Customer delivery of cryo-stored clones on dry ice (3 vials for the 1-3 selected clones) and 10 ml of cell culture supernatant/ cell line Polyclonal Antibody Development The standard service includes: 1. Discussion of the details of the project including features such as the type of immunogen, the final properties of the antibodies, and the species and number of animals used. In this context, the service offers the possibility to produce polyclonal antibodies in rabbits, rats, mice or hens. 2. Immunization protocol: the standard immunization protocol is performed in rabbits and it takes approximatelly 12 weeks and includes extraction of pre-immune serum, first injection with Complete adjuvant and boosting injections (3) every 21 days with icomplete adjuvant. There is a control bleed after 10 days of each immunization. The final blood extaction is performed on day 73. A customized protocol can be discussed with the customer. 3. ELISA testing (optional): samples obtained after each boosting injection and the final antisera will be tested against the antigen following the criteria agreed upon in the previous discussion with the costumer. The costumer will receive 50-80 mL of the final serum from each rabbit and about 5 mL samples of the pre-immune serum, and the partial blood extractions made during the immunization protocol. Additional services (optional): Preparation of bioconjugates: labelled antibodies, haptenized proteins and enzymes, biotinylated and fluorescent probes, gold nanoparticle conjugates, etc. In vitro mocolonal antibody production Antibody purification (affinity purification, protein G or A purification) Development of immunochemical methods: ELISA, immunoaffinity columns, etc. Antibody isotype determination Hybridoma cryopreservation. Mycoplasma testing
Contact Dr. Nuria Pascual. E-mail: nuria.pascual@iqac.csic.es Teléfono: 934006100 Ext. 5116
The Custom Antibody Service CAbS provides a high quality service, whereby every project is examined by the Scientific Committee which produces a feasibility report. Once is project is completed, the antibody generated is deleivered with a report including data related to its characterisation as well as aspects relating to its production. The service is facilitated with a laboratory for the development and productionof monoclonal antibodies which is kept under positive pressure to maintain sterile conditions. The laboratory is fully equipped for obtaining, selecting andstoring monoclonal antibody producing hybridomas. The equipment is composed of an inverted phase contrast microscope with a camer, two Class II laminar flow biological safety cabinets, two CO2 incubators, a bioreactor for hybridoma culture, and tangential filtration unit for purification of the antibodies produced in the culture. In addition, the facility has infrastructure for the production of polyclonal antibodies, as well as laboratories for the synthesis of bioconjugates, haptens, purification of antibodies and the performance of immunoassays.