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  - 
TY  - JOUR
A1  - Lütkecosmann, Steffi
A1  - Warsinke, Axel
A1  - Tschöpe, Winfried
A1  - Eichler, Rüdiger
A1  - Hanack, Katja
T1  - A novel monoclonal antibody suitable for the detection of leukotriene B4
JF  - Biochemical and biophysical research communications
N2  - Leukotriene B4 as an inflammatory mediator is an important biomarker for different respiratory diseases like asthma, chronic obstructive pulmonary disease or cystic lung fibrosis. Therefore the detection of LTB4 is helpful in the diagnosis of these pulmonary diseases. However, until now its determination in exhaled breath condensates suffers from problems of accuracy. Reasons for that could be improper sample collection and preparation methods of condensates and the lack of consistently assay specificity and reproducibility of the used immunoassay detection system. In this study we describe the development and the characterization of a specific monoclonal antibody (S27BC6) against LTB4, its use as molecular recognition element for the development of an enzyme-linked immunoassay to detect LTB4 and discuss possible future diagnostic applications.
KW  - Leukotriene B4
KW  - Monoclonal antibody
KW  - Immunosensor
KW  - Chronic obstructive pulmonary disease (COPD)
KW  - Hapten
Y1  - 2017
U6  - https://doi.org/10.1016/j.bbrc.2016.11.157
SN  - 0006-291X
SN  - 1090-2104
VL  - 482
IS  - 4
SP  - 1054
EP  - 1059
PB  - Elsevier
CY  - San Diego
ER  -