Computational protocol: The CD37-targeted antibody-drug conjugate IMGN529 is highly active against human CLL and in a novel CD37 transgenic murine leukemia model

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Protocol publication

[…] Flow experiments were performed using Beckman Coulter FC 500 flow cytometers (Brea, CA). Apoptosis assays were conducted on cells stained with fluorescein isothiocyanate (FITC) conjugated Annexin V and propidium iodide (PI) in 1X Annexin V binding buffer (BD Biosciences, San Jose, CA). Other experiments used fluorochrome-labeled monoclonal antibodies against mouse CD3 (17A2), B220 (RA3-6B2), CD19 (ID3), CD5 (53-7.3), CD4 (RM4-5), CD8 (53-6.7), CD45 (30-F11) (BD Biosciences) and anti-human CD37 (K7153A) conjugated to R-Phycoerythrin (K7153A-PE) that was provided by ImmunoGen, Inc. (Waltham, MA). Flow cytometric data was analyzed using Kaluza software (Beckman Coulter), with the exception of cell cycle experiments which were analyzed with Flowjo (Tree Star, Ashland, OR). Gating was verified with appropriate Fluorescence Minus One (FMO) controls. Absolute cell concentrations were obtained by quantitative flow cytometry using Count Bright absolute counting beads (Invitrogen). Quantification of CD37 surface expression was performed using the QuantiBRITE PE bead assay (BD Biosciences) and 1:1 conjugated K7153A-PE. […]

Pipeline specifications

Software tools Kaluza, FlowJo
Application Flow cytometry
Organisms Mus musculus, Homo sapiens
Diseases Hematologic Diseases, Leukemia, Lymphoma, Non-Hodgkin, Neoplasms, Leukemia, Lymphocytic, Chronic, B-Cell, Drug-Related Side Effects and Adverse Reactions