Journal: Communications Medicine

Article Title: Preclinical characterization of an active immunotherapy targeting calcitonin gene-related peptide

doi: 10.1038/s43856-025-00870-2

Figure Lengend Snippet: Proteome and peptide screens were used to evaluate potential off-target binding of affinity-purified p4796kb antibodies. a Manhattan plot displays results from the HuProt Human Proteome Microarray screen of antibody binding. Colors represent each block from the array, containing approximately 1000 proteins each. Spots represent individual protein intensity score from array. Hits are annotated with protein names. b Manhattan plot from peptide counter screen of 21 putative hits from proteome screen. Overlapping 15mer peptides were synthesized and printed on arrays. Spots represent individual peptide intensity scores from array. Peptide sequences of hits are displayed. Sera from guinea pigs immunized with p4796kb were tested for binding to putative hits from screens ( c ) and calcitonin family ( d ) measured by ELISA. Pre-immune sera were used as controls. Data are presented as means +/− SEM; n = 3. *** p < 0.0001.

Article Snippet: Binding of p4796kb-derived sera to 3 hits from the HuProt™ microarray screen (HSP90 beta protein, StressMarq Bioscience, SPR-102B; AKR1B10 protein, MyBioSource, MBS203315; PTPRD protein, Acro, PTD-H52H9) and to other propeptides that belong to the calcitonin/CGRP peptide family, including recombinant adrenomedullin (ADM, MyBioSource, MBS2012013), recombinant adrenomedullin 2 (ADM2, MyBioSource, MBS2123890), synthetic amylin (Abcam, ab142398), and recombinant calcitonin (Abcam, ab153793), were further evaluated.

Techniques: Binding Assay, Affinity Purification, Microarray, Blocking Assay, Synthesized, Enzyme-linked Immunosorbent Assay

Journal: Biochemical Journal

Article Title: On the function of TRAP substrate-binding proteins: the isethionate-specific binding protein IseP

doi: 10.1042/BCJ20240540

Figure Lengend Snippet: ( A ) Thermal shift heatmap for Oa IseP displaying changes in melting temperature ( Δ T m D , °C) with various phosphate and sulfur metabolites. The mean from technical quadruplicates of each condition is shown. Isethionate (position H10) was the only hit ( Δ T m D > 2°C) amongst 94 metabolites from the Phenotype MicroArray PM4A screen. These data are also tabulated in . ( B ) Oa IseP binding curve and affinity ( K D ) for isethionate determined by ITC. This experiment was conducted with technical triplicates and a representative result is shown. When error bars are not apparent, they are smaller than the data points. These data are also tabulated in .

Article Snippet: For experiments using the Phenotype MicroArray PM4A Biolog screen, the compounds were dissolved in 50 μl of 50 mM Tris pH 8.0, 150 mM NaCl buffer, before 7.5 μl was used in each replicate (Biolog does not disclose the exact concentrations in their screens).

Techniques: Microarray, Binding Assay

Journal: Biochemical Journal

Article Title: On the function of TRAP substrate-binding proteins: the isethionate-specific binding protein IseP

doi: 10.1042/BCJ20240540

Figure Lengend Snippet: Thermal stability and isothermal titration calorimetry for isethionate binding

Article Snippet: For experiments using the Phenotype MicroArray PM4A Biolog screen, the compounds were dissolved in 50 μl of 50 mM Tris pH 8.0, 150 mM NaCl buffer, before 7.5 μl was used in each replicate (Biolog does not disclose the exact concentrations in their screens).

Techniques: Isothermal Titration Calorimetry, Control, Binding Assay