Recombinant Mouse Anti-Lysozyme Antibody scFv Fragment (D1.3)

Figure 1 Representative images of spinal cord sections treated as in (A).

For each mAb, images with 8-18C5 or D1.3 pretreatment were acquired using the same settings and processed analogously. Alexa Fluor 555 and DAP1 fluorescence are pseudocolored red and green, respectively. Scale bar, 20 mm.

Bansal, P., Khan, T., Bussmeyer, U., Challa, D. K., Swiercz, R., Velmurugan, R., ... & Ward, E. S. (2013). The Encephalitogenic, Human Myelin Oligodendrocyte Glycoprotein–Induced Antibody Repertoire Is Directed toward Multiple Epitopes in C57BL/6-Immunized Mice. The Journal of Immunology, 191(3), 1091-1101.

Figure 2 A subset of the mAbs bind to mMOG expressed on the surface of transfected EL-4 cells.

EL4-MOG cells were incubated with 50 mg/ml mAb (black) or, as control, anti-lysozyme D1.3 Ab (mouse IgG1; gray). Bound Abs were detected using an Alexa 647–labeled anti-mouse IgG (H + L chain–specific) conjugate. Data are representative of three independent experiments.

Bansal, P., Khan, T., Bussmeyer, U., Challa, D. K., Swiercz, R., Velmurugan, R., ... & Ward, E. S. (2013). The Encephalitogenic, Human Myelin Oligodendrocyte Glycoprotein–Induced Antibody Repertoire Is Directed toward Multiple Epitopes in C57BL/6-Immunized Mice. The Journal of Immunology, 191(3), 1091-1101.

Figure 3 SPR assays to assess competition of mAbs with 8-18C5 for binding to recombinant mMOG.

mAb 8-18C5 and mMOG were immobilized on two flow cells of a CM5 sensor chip. mMOG (5 mg/ml; ∼200 nM) was injected (arrowhead) followed by mAb (arrow). Each mAb was injected at a concentration of ∼5-fold its Kd value for interaction with mMOG. Overlaid sensorgrams are shown for the 8-18C5 coupled flow cell (showing mMOG binding followed by mAb binding if the mAb does not compete with 8-18C5) and the mMOG coupled flow cell (showing only mAb binding as a control). The anti-lysozyme Ab D1.3 (35) was used as a control (bottom right panel). Data are representative of at least two injections for each mAb.

Bansal, P., Khan, T., Bussmeyer, U., Challa, D. K., Swiercz, R., Velmurugan, R., ... & Ward, E. S. (2013). The Encephalitogenic, Human Myelin Oligodendrocyte Glycoprotein–Induced Antibody Repertoire Is Directed toward Multiple Epitopes in C57BL/6-Immunized Mice. The Journal of Immunology, 191(3), 1091-1101.