af488 conjugated antibody  (Bioss)

Journal: bioRxiv

Article Title: CD38 biallelic loss is a recurrent mechanism of resistance to anti-CD38 antibodies in multiple myeloma

doi: 10.1101/2024.12.17.628799

Figure Lengend Snippet: Functional validation of CD38 variants. A) Wild-type and mutant CD38 protein expression in K562 cell lines by monoclonal and polyclonal anti-CD38 antibodies. B) CD38 binding assays: K562 cells were incubated with either Dara or Isa followed by secondary anti-IgG staining. C) Left: Repeated binding assay for K562 CD38 R140G (blue) at different drug concentrations with dose-independent decreased affinity for Dara compared to wild-type (grey). Right: Change in mean fluorescence intensity (MFI) recapitulating maintained binding of Isa for the R140G protein at all drug concentrations compared to the dose-independent decreased binding of Dara. D) Co-culture experiment with healthy donor peripheral blood mononuclear cells at Effector:Target ratio of 20:1 for various Dara and Isa concentrations demonstrating impaired antibody dependent cellular cytotoxicity of Dara compared to Isa for K562 CD38 R140G compared to K562 CD38 wild-type . E) Structure of human CD38 (grey) with mapped epitopes for Isa (blue) and Dara (red), and three identified missense SNV (green spheres). Protein structure was visualized using PyMOL (The PyMOL Molecular Graphics System, Version 3.0, Schrödinger, LLC). The effect of R140G is described in text. Cys 275 forms a critical disulfide bond stabilising the protein structure. Mutation to bulky Tyr destabilizes CD38 structural integrity, affecting both epitopes. The aliphatic Leu 153 contributes to CD38 hydrophobic core formation under the Isa epitope. Its mutation to a charged His will destabilize the Isa epitope structural integrity affecting the antibody binding.

Article Snippet: Surface expression of CD38 was confirmed using monoclonal (BioLegend, #356610) and polyclonal (Cell Signaling Technology, #14637S) anti-CD38 antibodies.

Techniques: Functional Assay, Mutagenesis, Expressing, Binding Assay, Incubation, Staining, Fluorescence, Co-Culture Assay

Journal: Molecular Imaging and Biology

Article Title: Development of New CD38 Targeted Peptides for Cancer Imaging

doi: 10.1007/s11307-024-01901-5

Figure Lengend Snippet: Radiolabeling and cellular uptake studies ( a ) Representative chromatograph of the 64 Cu labeled CD38-peptide (NODAGA-PEG4-SL022-GGS) under two different conditions; ( b ) In vitro blocking study of 64 Cu-NODAGA-PEG4-SL022-GGS in MM.1S-GFP-CBR-WT cells with unlabeled NODAGA-PEG4-SL022-GGS, daratumumab and isatuximab respectively.

Article Snippet: Nonspecific binding was blocked by incubating cells with 1% BSA, 0.1% Tween 20 in DPBS for 30 min. After overnight incubation at 4 °C with anti-CD38 mouse polyclonal antibody (# PA5-95,840, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:100 dilution) or anti-CD31 mouse monoclonal antibody (JC/70A #MA5-13,188, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:50 dilution), cells were washed and incubated for 1 h at room temperature with Alexa Fluor 594- or 647-conjugated secondary goat anti-mouse IgG1 (cross-adsorbed, # A-21240, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:50 dilution) or F(ab')2-goat anti-rabbit IgG (H + L) (cross adsorbed, # A48285, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:100 dilution).

Techniques: Radioactivity, Labeling, In Vitro, Blocking Assay

Journal: Molecular Imaging and Biology

Article Title: Development of New CD38 Targeted Peptides for Cancer Imaging

doi: 10.1007/s11307-024-01901-5

Figure Lengend Snippet: a CD38 & CD31 expression in MM.1S-GFP-CBR-WT cells. CD38 (left column): Immunocytochemistry analysis of CD38 expression in MM.1S-CBR-GFP cells with the CD38 1° antibody followed by the AlexaFluor 647 conjugated goat anti-rabbit IgG secondary antibody. Nuclei were counter stained with Hoechst 33,342. The sections were imaged with GFP filter pair (410/520) for endogenous GFP expression, Cy5 (620/705) for CD38 with AlexaFlour 647-conjugated secondary antibody, and Hoechst 33,342 (370/405) to stain nuclei. CD31 (right column): Immunohistochemistry analysis of CD31 expression in MM.1S-CBR-GFP cells with CD31 antibody followed with AlexaFluor 647 conjugated goat anti-rabbit IgG secondary antibody. Nuclei were counter-stained with Hoechst 33,342. The sections were imaged with GFP filter pair (410/520) to visualize endogenous GFP expression, Cy5 (620/705) for CD31 expression with AlexaFlour 647-conjugated secondary antibody, Hoechst 33,342 (370/405) to stain nuclei; b Characterization of the disseminated MM.1S-CBR-GFP-WT tumor model. In vivo BLI showing presence of disseminated MM.1S-CBR-GFP-WT in both femurs and tibia at 5 weeks after tumor implantation; c MM.1S-CBR-GFP-WT tumor burden additionally evidenced in the femur via GFP intracellular expression seen with epifluorescence microscopy of unstained femur section, scale bar – 20 µm; d Histological assessment of the femur with H&E staining shown at 10x, 20x, 40 × at scale of 200 µm, 100 µm, 20 µm, respectively.

Article Snippet: Nonspecific binding was blocked by incubating cells with 1% BSA, 0.1% Tween 20 in DPBS for 30 min. After overnight incubation at 4 °C with anti-CD38 mouse polyclonal antibody (# PA5-95,840, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:100 dilution) or anti-CD31 mouse monoclonal antibody (JC/70A #MA5-13,188, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:50 dilution), cells were washed and incubated for 1 h at room temperature with Alexa Fluor 594- or 647-conjugated secondary goat anti-mouse IgG1 (cross-adsorbed, # A-21240, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:50 dilution) or F(ab')2-goat anti-rabbit IgG (H + L) (cross adsorbed, # A48285, Invitrogen, ThermoFisher Scientific, Waltham, MA, 1:100 dilution).

Techniques: Expressing, Immunocytochemistry, Staining, Immunohistochemistry, In Vivo, Tumor Implantation, Epifluorescence Microscopy

Journal: Cell Reports Medicine

Article Title: Boosting NAD preferentially blunts Th17 inflammation via arginine biosynthesis and redox control in healthy and psoriasis subjects

doi: 10.1016/j.xcrm.2023.101157

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CD38 , Cell Signaling , Cat# 92457; RRID: N/A.

Techniques: Flow Cytometry, Virus, Recombinant, Staining, Cell Stimulation, CyQUANT Assay, Proliferation Assay, Bicinchoninic Acid Protein Assay, In Vitro, Antioxidant Assay, Detection Assay, GSH Assay, Bioassay, Transcription Factor Assay, Extraction, Enzyme-linked Immunosorbent Assay, Control, Software, Cell Isolation, Transfection