Recombinant Human Anti-IFN-gamma Antibody scFv Fragment (HuZAF) (CAT#: PABX-113-S (P))

Figure 1 Competitive binding analysis of AF2 and HuZAF.

A, Graph represents the relative ability of AF2 and HuZAF to compete with bAF2 for binding to human IFN-g in an ELISA format. Increasing concentrations of either AF2 (F) or HuZAF (■) were added to wells coated with human IFN-g, after which bAF2 was added. Error bars indicate the SE of triplicate samples. B, Neutralization of IFN-g-mediated induction of MHC class II
molecule expression on Hs294T cells. Graph represents the median fluorescent intensity of HLA-DR expression on Hs294T cells incubated with IFN-g and increasing concentrations of either AF2 (F) or HuZAF (■).
Median fluorescent intensity of Hs294T cells incubated in the absence of IFN-g was 3.74; median fluorescent intensity in the presence of IFN-g, but in the absence of Ab was 9.65.

Landolfi, N. F., Thakur, A. B., Fu, H., Vásquez, M., Queen, C., & Tsurushita, N. (2001). The integrity of the ball-and-socket joint between V and C domains is essential for complete activity of a humanized antibody. The Journal of Immunology, 166(3), 1748-1754.

Figure 2 Involvement of AF2 VH residues 11 and 38 in neutralization of IFN-g activity.

Graph represents the median fluorescent intensity of HLA-DR expression on Hs294T cells incubated with IFN-g and increasing concentrations of either EuAF2 (F), or EuAF2 with the following substitutions: R38K (l), V11L (triangles), or V11L, R38K (■).

Landolfi, N. F., Thakur, A. B., Fu, H., Vásquez, M., Queen, C., & Tsurushita, N. (2001). The integrity of the ball-and-socket joint between V and C domains is essential for complete activity of a humanized antibody. The Journal of Immunology, 166(3), 1748-1754.

Figure 3 Stereo illustration of the three-dimensional structure of the HuZAF Fab, prepared with the program MOLSCRIPT (Kraulis, 1991).

Secondary elements are superimposed on backbone tracings, which are colored yellow in the L chain and steel blue in the H chain. -Strands are represented by directional arrows: cyan for the four-stranded -pleated sheets and magenta for the ®ve-stranded (VL) and three-stranded layers (CL) of the L chain and red and white for the corresponding -pleated sheets in the H chain. Helices are identi®ed by red and yellow spirals. CDRs are marked 1, 2 and 3.

Bourne, P. C., Terzyan, S. S., Cloud, G., Landolfi, N. F., Vásquez, M., & Edmundson, A. B. (2004). Three-dimensional structures of a humanized anti-IFN-γ Fab (HuZAF) in two crystal forms. Acta Crystallographica Section D: Biological Crystallography, 60(10), 1761-1769.