TY  - JOUR
A1  - Holzlöhner, Pamela
A1  - Hanack, Katja
T1  - Generation of murine monoclonal antibodies by hybridoma technology
JF  - JoVE : Video journal
N2  - Monoclonal antibodies are universal binding molecules and are widely used in biomedicine and research. Nevertheless, the generation of these binding molecules is time-consuming and laborious due to the complicated handling and lack of alternatives. The aim of this protocol is to provide one standard method for the generation of monoclonal antibodies using hybridoma technology. This technology combines two steps. Step 1 is an appropriate immunization of the animal and step 2 is the fusion of B lymphocytes with immortal myeloma cells in order to generate hybrids possessing both parental functions, such as the production of antibody molecules and immortality. The generated hybridoma cells were then recloned and diluted to obtain stable monoclonal cell cultures secreting the desired monoclonal antibody in the culture supernatant. The supernatants were tested in enzyme-linked immunosorbent assays (ELISA) for antigen specificity. After the selection of appropriate cell clones, the cells were transferred to mass cultivation in order to produce the desired antibody molecule in large amounts. The purification of the antibodies is routinely performed by affinity chromatography. After purification, the antibody molecule can be characterized and validated for the final test application. The whole process takes 8 to 12 months of development, and there is a high risk that the antibody will not work in the desired test system.
KW  - Immunology
KW  - Issue 119
KW  - monoclonal antibodies
KW  - hybridoma technology
KW  - myeloma cells
KW  - B lymphocytes
KW  - antigen
KW  - immunconjugate
Y1  - 2017
U6  - https://doi.org/10.3791/54832
SN  - 1940-087X
IS  - 119
PB  - JoVE
CY  - Cambridge
ER  -