A Novel Anti-Histone H1 Monoclonal Antibody, SSV Monoclonal Antibody, Improves Lung Injury and Survival in a Mouse Model of Lipopolysaccharide-Induced Sepsis-Like Syndrome

Figure 1

Characterization of SSV mAb. (a) The binding of SSV mAb to KLH-conjugated SSV peptide (KLH-SSV) was determined by ELISA. Native KLH (KLH) was used as a control to eliminate nonspecific binding. Increasing SSV mAb concentrations were added to the wells of a microtiter plate that had been coated with native KLH or KLH-conjugated SSV. After washing and binding of an HRP-conjugated secondary antibody, the amount of SSV mAb was determined by a colorimetric HRP activity assay at 405 nm. (b) SSV mAb binds to histone H1. Increasing SSV mAb concentrations were added to the wells of a microtiter plate that had been coated with histone H1 or BSA as control. After washing and binding of an HRP-conjugated secondary antibody, the amount of SSV mAb was determined as described above. (c) SSV mAb binding to KLH-conjugated SSV was significantly inhibited () by histone H1. In these assays, KLH-conjugated SSV (c) was used to coat the wells of a microtiter plate. SSV mAb was incubated with increasing concentrations of the histones shown in the respective panels, and the SSV mAb-histone mixtures were added to the coated wells blocked with the blocking solution. After washing, the wells were treated with secondary antibody, and the amount of bound SSV mAb in each well was determined as described above. ^*Statistically significant difference ().

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