Ox40 fc

Example 10

For ELISA assays assessing functional binding of OX40L to its corresponding receptor, coating of microtiter plates was performed with 1 μg/ml OX40-Fc (Bio-Techne GmbH, Wiesbaden-Nordenstadt, Germany). After blocking with StartingBlock (Life Technologies GmbH, Darmstadt, Germany), wells were incubated with indicated concentrations of GITRL compound of different configuration (Protein A, Protein B and Protein C). GITRL bound to its corresponding receptor was detected via its Strep Tag II employing the anti-StrepTag-peroxidase StrepTactin-HRP (1:5000, IBA GmbH, Goettingen, Germany) and subsequent detection of the converted Peroxidase-substrate TMB one (Kem-En-Tec Diagnostics, Taastrup, Denmark) at a wavelength of 450 nm in an ELISA reader. FIG. 6 clearly demonstrates functional binding of all tested protein configurations of the invention. Binding capacity of N- and C-terminal shortened variants is comparable to Protein A.

Example 10

For ELISA assays assessing functional binding of OX40L to its corresponding receptor, coating of microtiter plates was performed with 1 μg/ml OX40-Fc (Bio-Techne GmbH, Wiesbaden-Nordenstadt, Germany). After blocking with StartingBlock (Life Technologies GmbH, Darmstadt, Germany), wells were incubated with indicated concentrations of GITRL compound of different configuration (Protein A, Protein B and Protein C). GITRL bound to its corresponding receptor was detected via its Strep Tag II employing the anti-StrepTag-peroxidase StrepTactin-HRP (1:5000, IBA GmbH, Goettingen, Germany) and subsequent detection of the converted Peroxidase-substrate TMB one (Kem-En-Tec Diagnostics, Taastrup, Denmark) at a wavelength of 450 nm in an ELISA reader. FIG. 6 clearly demonstrates functional binding of all tested protein configurations of the invention. Binding capacity of N- and C-terminal shortened variants is comparable to Protein A.