Journal: PLoS Neglected Tropical Diseases

Article Title: The OmpL37 Surface-Exposed Protein Is Expressed by Pathogenic Leptospira during Infection and Binds Skin and Vascular Elastin

doi: 10.1371/journal.pntd.0000815

Figure Lengend Snippet: Binding of OmpL37 (concentration ranging from 0 to 2 µM) to 1 µg of immobilized (A) human skin elastin, (B) human aorta elastin, (C) human plasma fibrinogen, (D) human plasma fibrinogen fragment D, (E) human plasma fibronectin, and (F) murine laminin was measured by ELISA. The mean optical density at 450 nm ± the standard deviation of three independent experiments is shown at each point. The apparent K d for saturating binding was estimated as the concentration of recombinant OmpL37 resulting in half-maximal binding (see text and ). OmpL36 served as a negative control.

Article Snippet: Host ligands included human plasma fibronectin (Sigma-Aldrich), human plasma fibronectin 30-kDa proteolytic fragment (heparin-binding domain, Sigma-Aldrich), human plasma fibronectin 45-kDa proteolytic fragment (gelatin-binding domain, Sigma-Aldrich), human fibroblast fibronectin (Calbiochem, La Jolla, CA), human plasma fibrinogen (HYPHEN BioMed, France), human plasma fibrinogen fragment D (HYPHEN BioMed), murine laminin (Sigma-Aldrich), bovine skin collagen type I (Sigma-Aldrich), human placenta collagen type III (Sigma-Aldrich), human placenta collagen type IV (Sigma-Aldrich), soluble human skin elastin (Elastin Products Company, Owensville, MO), soluble human aorta elastin (Sigma-Aldrich), bovine kidney heparan sulfate (Sigma-Aldrich), shark cartilage chondroitin sulfate (Sigma-Aldrich), fetal calf serum fetuin (Sigma-Aldrich), and bovine serum albumin (BSA, Sigma-Aldrich).

Techniques: Binding Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Recombinant, Negative Control

Journal: PLoS Neglected Tropical Diseases

Article Title: The OmpL37 Surface-Exposed Protein Is Expressed by Pathogenic Leptospira during Infection and Binds Skin and Vascular Elastin

doi: 10.1371/journal.pntd.0000815

Figure Lengend Snippet: Microtiter wells were coated with 1 µg of human skin elastin and binding was measured by ELISA. (A) PBS-5 mM MgCl 2 (no cells) or 1×10 8 L. interrogans L1-130 in PBS-5 mM MgCl 2 (cells) without antiserum (no Ab) or in the presence of OmpL37 or OmpL54 antiserum (Ab) diluted 1∶40, 1∶80, 1∶160, 1∶320, and 1∶1280 were added to the elastin-coated wells. (B) Recombinant OmpL37 (0.5 µg) was preincubated with or without the IgG purified from 10 times diluted OmpL37 or OmpL47 antiserum at 1∶40, 1∶160, and 1∶640 dilution and the OmpL37 or OmpL47 antiserum at 1∶2500 dilution prior to addition to elastin-coated wells. Mean absorbance at 450 nm ± the standard deviation of a representative experiment performed in triplicate is shown. Statistical significance was evaluated by one-way ANOVA comparing leptospiral or recombinant OmpL37 binding in the presence of antibodies compared with binding in the absence of antibodies (** P<0.001, * P<0.05).

Article Snippet: Host ligands included human plasma fibronectin (Sigma-Aldrich), human plasma fibronectin 30-kDa proteolytic fragment (heparin-binding domain, Sigma-Aldrich), human plasma fibronectin 45-kDa proteolytic fragment (gelatin-binding domain, Sigma-Aldrich), human fibroblast fibronectin (Calbiochem, La Jolla, CA), human plasma fibrinogen (HYPHEN BioMed, France), human plasma fibrinogen fragment D (HYPHEN BioMed), murine laminin (Sigma-Aldrich), bovine skin collagen type I (Sigma-Aldrich), human placenta collagen type III (Sigma-Aldrich), human placenta collagen type IV (Sigma-Aldrich), soluble human skin elastin (Elastin Products Company, Owensville, MO), soluble human aorta elastin (Sigma-Aldrich), bovine kidney heparan sulfate (Sigma-Aldrich), shark cartilage chondroitin sulfate (Sigma-Aldrich), fetal calf serum fetuin (Sigma-Aldrich), and bovine serum albumin (BSA, Sigma-Aldrich).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Purification, Standard Deviation