Mab006

PAM3CSK4 is a synthetic diacylated lipopeptide activating TLR-2 with cooperation from TLR-1. PAM3CSK4 (InvivoGen, San Diego, CA) was dissolved in sterile endotoxin free water to a final concentration of 0.5 mg/mL. Mice were injected intraperitoneally (i.p.) with 50 μg of PAM3CSK4 (henceforth referred to as PAM) solution or control [sterile endotoxin free water or sterile phosphate buffered saline (PBS)]. In time course studies of laser-induced CNV area, a single dose of PAM was injected at either day 0, 1, 2, 3, or 4 relative to laser application. For subsequent studies, single i.p. injections of PAM or vehicle control (either sterile water or PBS) were administered 2 days after laser application.
4G3 is a human Fab converted to a full-length IgG antibody with a mouse IgG1 Fc tail.^{38 (link)} 4G3 binds to mouse VEGF₁₆₄ with a dissociation constant of 10 pM and neutralizes mouse VEGF binding to human VEGFR2 with an EC₅₀ of 0.15 nM in a binding assay (ELISA; MSD, Rockville, MD).^{38 (link)} For efficacy studies in the mouse laser CNV assay, 4G3 dissolved in PBS was dosed i.p. at doses of 0.1, 0.3, 1.0, 3.0, or 10 mg/kg at day 0, 2, and 4 after laser. Either rat IgG_2a isotype (MAB006; R&D systems, McKinley Place, MN) dissolved in PBS or PBS was dosed i.p. as negative controls for laser CNV studies.