TY  - JOUR
A1  - Messerschmidt, Katrin
A1  - Heilmann, Katja
T1  - Toxin-antigen conjugates as selection tools for antibody producing cells
JF  - Journal of immunological methods
N2  - The generation of antibodies with designated specificity requires cost-intensive and time-consuming screening procedures. Here we present a new method by which hybridoma cells can be selected based on the specificity of the produced antibody by the use of antigen-toxin-conjugates thus eliminating the need of a screening procedure. Initial experiments were done with methotrexate as low molecular weight toxin and fluorescein as model antigen. Methotrexate and a methotrexate-fluorescein conjugate were characterized regarding their toxicity. Afterwards the effect of the fluorescein-specific antibody B13-DE1 on the toxicity of the methotrexate-fluorescein conjugate was determined. Finally, first results showed that hybridoma cells that produce fluorescein specific antibodies are able to grow in the presence of fluorescein-toxin-conjugates.
KW  - Monoclonal antibody
KW  - Hybridoma technology
KW  - Selection of antibody producing cells
Y1  - 2013
U6  - https://doi.org/10.1016/j.jim.2012.10.010
SN  - 0022-1759
VL  - 387
IS  - 1-2
SP  - 167
EP  - 172
PB  - Elsevier
CY  - Amsterdam
ER  - 
TY  - JOUR
A1  - Michelchen, Sophia
A1  - Micheel, Burkhard
A1  - Hanack, Katja
T1  - In vitro immunization approach to generate specific murine monoclonal IgG antibodies
JF  - Journal of immunological methods : JIM
N2  - Generating a monoclonal antibody to date is a time intense process that requires immunization of laboratory animals. The transfer of the humoral immune response into in vitro settings enables a shortening of this process and circumvents the necessity of in vivo immunization. However, to orchestrate the complex interplay of dendritic cells, T and B lymphocytes in vitro is very challenging. We therefore aimed for a simplified approach focusing on the protagonist of antibody production: the B lymphocyte. We activated purified murine B lymphocytes alone in vitro by using combinations of antigen and stimuli. We were able to induce a specific antibody response within ten days of culture against a viral coat protein as model antigen. Antibodies were of both IgM and IgG subclass. The stimulated B lymphocytes were transformed into permanently antibody-producing hybridomas by cell fusion technology. We furthermore used this method to induce a specific antibody response against L. pneumophila in vitro. We thus established a useful and effective in vitro protocol to generate monoclonal antibodies. By overcoming the necessity of in vivo immunization this protocol may be the first step towards a universal strategy to generate antibodies from various species.
KW  - Monoclonal antibody
KW  - Hybridoma technology
KW  - In vitro immunization
KW  - B cell activation
Y1  - 2021
U6  - https://doi.org/10.1016/j.jim.2021.113149
SN  - 0022-1759
SN  - 1872-7905
VL  - 499
PB  - Elsevier
CY  - Amsterdam
ER  -