Production of policlonal and monoclonal antibodies

Service description

The service, which adapts to needs of each client, can produce antibodies against proteins, peptides, organic molecules and other antigens, through standardized or personalized protocols and with special emphasis on the design of the immunogens, which is essential to modulate the selectivity and affinity of the antibody. It is also one of the few laboratories that offers advice and support in relation to the preparation of immunogens and the production of antibodies for low molecular weight molecules such as pigments, hormones or anabolic steroids or peptides. The immunogen can be provided by the user or can be designed and synthesized or bionconjugated by the same antibody production service. The protocols and the care of animals are performed according to the specifications of the Real Decreto 53/2013 about ¿Protection of the animals used for experimentation and other scientific activities¿, both for monoclonal and polyclonal antibody production. Monoclonal Antibody Development and Production: The Monoclonal Antibody Production service is based on Balb/c mice and includes the following phases: -PRELIMINARY: Discussion of the details of the project (screening system, selection criteria, special requirements), planning and work preparation. -IMMUNIZATION, CELLULAR FUSION, SELECTION AND SCREENING: Immunization of at least 4 mice (Balb-c.), cell culturing of myeloma cells of the P3x63Ag8.653 cell line, cellular fusion, selection and screening. -CLONING: Cloning, screening and cultivation of positive hybridomas. The user will receive, if they wish, the supernatant of the necessary culture for each positive clone to test using their own methods. -CRYOPRESERVATION: Three vials will be cryopreserved for each stable positive clone. The cryovials and 10 mL of supernatant of the culture of each positive clone will be sent to the client. -CARACTERIZATION OF THE SELECTED CLONES: The Ig isotype will be determined and detection of mycoplasm test will be performed. The corresponding report will be provided. Hibridoma cell line are used to the ¿in vitro¿ MONOCLONAL ANTIBODY PRODUCTION at a small or medium scale, by using cell bioreactors. Production of polyclonal antibodies: The service has a standard immunization protocol that includes monthly inoculations with Freund adjuvant for a period of 4 months. This protocol adapts to the needs of the required antiserums, keeping in mind the characteristics of the immunogen and the required characteristics of the antibody. To determine the progress of the immune response, a bleed is taken 10 days after the second immunization to obtain antiserum and the titer is determined through ELISA. Controls include the analysis of the preimmune serum obtained from a bleed performed prior to immunization. At the end of the process, the exsanguination of the animal is made using a surgical process under anesthesia. Normally 50-70 mL of hyperimmune serum is obtained per animal. Adicionales services - Antibody purification - Isotyping of obtained monoclonal antibodies - Conjugation of the antibodies or antigens - Storage of hybridomas in liquid N2 - Immunoassays