Journal: Pharmaceuticals

Article Title: Lactoferrin Binds through Its N-Terminus to the Receptor-Binding Domain of the SARS-CoV-2 Spike Protein

doi: 10.3390/ph17081021

Figure Lengend Snippet: Effect of hLF and bLF on S-protein binding. ACE2 ( A ), HSPG ( B ), and HAS ( C ) were coated on wells of a 96-well plastic plate. Casein was used as a negative control. After blocking, the wells were incubated overnight at 4 °C with the S-protein in the absence or presence of hLF or bLF at indicating concentrations (from 5 to 20 µg/mL). Afterwards, the wells were washed and the bound material was analyzed by immunoblotting using mAb to the S-protein. ( D ) Densitometric evaluation of at least three independent experiments. The values corresponding to the binding mixture ( input ) were set as 100%. The values corresponding to the casein binding were subtracted from the values of the respective bound protein bands; the resultant values are expressed as percentages of the inputs: std., molecular weight standard. The data from at least three blots were evaluated. Values of p * < 0.05 (as indicated) were considered to be significant or highly significant, respectively; std. corresponds to the lane with the molecular weight standards.

Article Snippet: For the binding assay, various molecules (ACE2, HSPG, HSA, hLF, casein) solubilized in PBS (pH 8.7) at a concentration of 5 µg/mL were coated on wells of a 96-well TPP plate (#92696, Sigma) for 2 h at 37 °C.

Techniques: Protein Binding, Negative Control, Blocking Assay, Incubation, Western Blot, Binding Assay, Molecular Weight

Journal: Pharmaceuticals

Article Title: Lactoferrin Binds through Its N-Terminus to the Receptor-Binding Domain of the SARS-CoV-2 Spike Protein

doi: 10.3390/ph17081021

Figure Lengend Snippet: Mapping of the S-protein binding to LF. hLF was coated on wells of a 96-well plastic plate. Casein was used as a negative control. After blocking, the wells were incubated overnight at 4 °C with the S-protein ( A ) or RBD ( C ) in the absence or presence of the peptides—pLF1, pLF3, pLF2, and pCTR, at indicated concentrations (from 5 to 20 µg/mL). Afterwards, the wells were washed and the bound material was analyzed by immunoblotting using mAb to S-protein. ( B , D ) Densitometric evaluations of at least three independent experiments. The values corresponding to the binding mixture ( input ) were set as 100%. The values corresponding to the casein binding were subtracted from the values of the respective bound protein bands; the resultant values are expressed as percentages of the inputs. The data from at least three blots were evaluated; values of p * < 0.05 (as indicated) were considered to be significant or highly significant, respectively.

Article Snippet: For the binding assay, various molecules (ACE2, HSPG, HSA, hLF, casein) solubilized in PBS (pH 8.7) at a concentration of 5 µg/mL were coated on wells of a 96-well TPP plate (#92696, Sigma) for 2 h at 37 °C.

Techniques: Protein Binding, Negative Control, Blocking Assay, Incubation, Western Blot, Binding Assay