Custom Antibody Development
Hybridoma development for new antibody producing cells
Biometec offer a step by step service for generation of monoclonal antibody producing cell lines to make it easy for you to plan the experiments and save costs. Together we can start and stop the antibody development after each step and you have to pay only the finished step.
The standard protocol can be adapted at customers option.
Standard protocol
Step 1 - Immunisation of 3 BALB/c mice with your antigen and standard protocol
Sera from mice will be tested by ELISA using your antigen. Once an acceptable titre is obtained cell fusion will be done using splenocytes from mouse with the best titre. Alternatively you can try an in vitro immunisation of spleen cells (for toxic antigens).
Antigen requirements: 2 mg for immunisation and 3-5 mg for screening
Time 4-8 weeks
Price 1000 Euro including fusion preparation and veterinary licence
Step 2 - Hybridoma generation and screening of specific clones
Splenocytes from spleen of the best mouse are fused with mouse myeloma cell line. The fused cells are plated into five 96well plates. After hybrid selection wells with growth are screened for specific antibody production by ELISA two times. The antibody positive clones are isolated and safety cryoconservated one time. Now the best stable 10-15 antibody producing primer cultures will be cryopreservated 5 times.
Time 3-4 weeks
Price 2000 Euro
Step 3 - Subcloning
Positive wells are subcloned three times by limiting dilution method and tested in each time by antigen specific ELISA.
Time 4-6 weeks
Price 2000 Euro for 5 clones
Step 4 - Expansion and cryopreservation
Test for recultivation, Isotyping, mycoplama testing
Time 2-4 weeks
Price 1000 Euro for 5 clones
References
Walschus,U. Witt.S. Wittmann Ch. Development of Monoclonal Antibodies Against Cry1Ab Protein from Bacillus thuringiensis and Their Application inan ELISA for detection of Transgenic Bt-Maize Taylor & Francis 2002,4 : 231-240
Kato,A., Ogasdawara, T., Homma, T., Saito, H., Matsumoto, K. Lipopolysaccaride-Binding protein critically regulates Lipopolysaccaride-induced IFN-ß Signaling pathway in human monocytes, J. Immunol. 2004, 172: 6185-6194
Bettina Albrecht, Stefan Hartwich, Zhenzhen Wen, Christine Wittmann,Fischallergennachweis in Lebensmitteln mit impedimetrischen Biosensoren, Labor More 3 / 2010
James B. Delehanty1 , Brandy J. Johnson , Thomas E. Hickey , Thomas Pons and Frances S. Ligler Binding and Neutralization of Lipopolysaccharides by Plant Proanthocyanidins J. Nat. Prod., 2007, 70 (11), pp 1718-1724
