For in vivo studies, T-DM1 (Genentech) was administered i.p. at 10 mg/kg once per week. ABT-737 (AbbVie) was administered i.p. at 70 mg/kg once per day. ABT-263 (AbbVie) was administered p.o. at 70 mg/kg once per day. T-DM1 was prepared according to Genentech recommendations in sterile water for injection (Gibco). ABT-737 and ABT-263 were prepared according to AbbVie recommendations. ABT-737 in 30% propylene glycol (Sigma) plus 0.5% Tween 80 (Sigma) and 65% D5W (5% dextrose in water, Sigma) pH 3–4 and ABT-263 in 60% PHOSAL 50 PG (Lipoid) plus 30% polyethylene glycol 400 (Dow Chemical) and 10% ethanol. Matched control animals received vehicle alone in the same manner as drug-treated counterparts. All animals were randomized into groups and weighed before treatment. Individual weights were used for dose calculations. Weights were re-measured at the 14-day experimental end point. Weight reductions >20% prevented continuous treatments beyond 14 days.
T dm1
Manufactured by Genentech
Sourced in United States
T-DM1 is a type of antibody-drug conjugate (ADC) used in laboratory research. It is composed of the anti-HER2 monoclonal antibody trastuzumab and the cytotoxic agent DM1 (emtansine). T-DM1 acts by binding to HER2-expressing cells and delivering the DM1 payload to induce cell death.
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Lab products found in correlation
Trastuzumab, by Genentech (3 mentions)
Hcc1954, by American Type Culture Collection (2 mentions)
Celltiter glo luminescent cell viability assay, by Promega (1 mentions)
Microplate reader, by Tecan (1 mentions)
Polyethylene glycol 400, by Dow (1 mentions)
Abt263, by Lipoid (1 mentions)
Propylene glycol, by Merck Group (1 mentions)
Phosal 50 pg, by Lipoid (1 mentions)
Abt 737, by Merck Group (1 mentions)
Abt 737, by Abbvie (1 mentions)
Tween 80, by Merck Group (1 mentions)
Abt 263, by Abbvie (1 mentions)
Rapamycin, by Merck Group (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Apoptosis detection kit, by Dojindo Laboratories (1 mentions)
Z vad fmk, by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
N acetyl l cysteine, by Merck Group (1 mentions)
Gne 317, by Genentech (1 mentions)
Attune flow cytometer, by Thermo Fisher Scientific (1 mentions)
13 protocols using t dm1
1
In vivo Xenograft Treatment Protocol
2
Cytotoxicity Assessment of Drug-Loaded NTFs
3
Trastuzumab and T-DM1 Drug Preparation
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Trastuzumab and vinorelbine were kindly provided by inpatient pharmacy. T-DM1 was supplied from Genentech Inc. (South San Francisco, CA) through a Materials Transfer Agreement. For the in vitro experiments, stock solutions of drugs were prepared in distilled water, stored at 4°C and diluted in fresh medium for use, whereas for the in vivo experiments trastuzumab and T-DM1 were daily dissolved in sterile saline solution (NaCl 0.9%).
4
Apoptosis detection in T-DM1 treated cells
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T-DM1 was purchased from Genentech Roche (San Francisco, CA, USA). Apoptosis Detection Kit was from Dojindo (Tokyo, JPN). Z-VAD-fmk, rapamycin, LY294002 and chloroquine (CQ) were obtained from Sigma-Aldrich (St Louis, MO, USA).
5
Dihydroartemisinin and T-DM1 Combination Therapy
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Dihydroartemisinin was from Selleckem (Munich, D) T-DM1 was provided by Genentech (South San Francisco, CA, USA). N-Acetyl-L- Cysteine (NAC) by Sigma-Aldrich, St. Louis, MO, USA.
6
Zirconium-89 Labeled Antibodies in HER2 Transgenic Mice
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Trastuzumab, T-DM1, muMAb 4D5 (the murine parent molecule of Trastuzumab), control antibodies, and GNE-317 were from Genentech, Inc. 89Zr-labeled antibodies were synthesized as described [19 (link)]. Anti-STEAP1 was used as the isotype-matched control antibody for Trastuzumab, and anti-CD22-DM1 served as the non-targeted control antibody–drug conjugate (ADC) for T-DM1. MMTV-human HER2 transgenic mice were established previously [20 (link)]. Tumors were obtained from the Fo2-1282 and Fo5 human HER2 transgenic lines for model development.
7
Characterization of Breast Cancer Cell Subpopulations
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Cells were either mechanically or enzymatically detached using Accutase (PAA), blocked and stained with 10 μg/ml Trastuzumab or 10 μg/ml T-DM1 (Genentech, Burlingame, CA, USA) followed by a Cy5-conjugated goat anti-human IgG (Rockland Immunochemicals, Gilbertsville, PA, USA) detection antibody. Simultaneously, CD44-RPE (Clone 2BJ18, BioLegend, San Diego, CA, USA), CD24-FITC (clone SWA-11, kindly provided by Professor Peter Altevogt, German Cancer Research Centre, Heidelberg, Germany) and the life stain 7-aminoactinomycin D (Sigma) were applied and analyzed on a FACSCalibur or an Attune flow cytometer (Life Technologies, Darmstadt, Germany). For E-Cadherin surface staining, a PerCP/Cy5.5-labeled anti-human CD324/E-Cadherin antibody (Clone 67A4) and a corresponding isotype control (Clone MOPC-21), both from BioLegend, were used. Where appropriate, expression levels are indicated as specific fluorescence intensity values, which are obtained by dividing the fluorescence intensity detected with the specific antibody by the signal measured with the isotype-matched control antibody. For fluorescence-activated cell sorting, the stained cells were separated twice on a Digital FACSVantage (BD Biosciences), first in yield and then in purity mode.
8
Cell line maintenance and authentication
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Myc B was purchased from Wako Chemicals (USA) and stored at −20 °C in DMSO. Trastuzumab (TZ) and T-DM1 were provided by Genentech. HEK293T, SKOV3 and HCC1954 cell lines were obtained from American Type Culture Collection, and authenticated by STR profiling. All cell lines were maintained in Dulbecco Modified Eagle Medium (DMEM, Multicell) supplemented with 10% fetal bovine serum (FBS, Multicell) and 1% antibiotics-antimycotic (Multicell), and cultured in a humidified incubator at 37 °C with 5% CO2.
9
Antibody-Drug Conjugate Preparation
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IgG1 trastuzumab (commercial name: Herceptin ® ) and rituximab (commercial name: Rituxan ® ) were purchased from Roche Pharmaceutical Company (Switzerland). Human IgG1 infliximab (commercial name: Humira ® ) was purchased from Sigma-Aldrich (USA). MC-VC-MMAE (CAS#: 646502-53-6) was purchased from NJ Biopharmaceuticals LLC (USA). SMCC-DM1 (CAS#: 1228105-51-8) and MC-MMAF (CAS#: 1228105-51-8) were purchased from Abzena (USA). T-DM1 (commercial name: Kadcyla ® ) was acquired from Genentech (USA) and reconstituted to 5 mg/mL formulation buffer (20 mM histidine containing 5% trehalose, pH 5.2) by gel-filtration. All other chemical reagents were acquired from Sigma-Aldrich (USA).
10
Evaluating T-DM1 and S-methyl DM1 Efficacy
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T-DM1 was kindly provided by Genentech and S-methyl DM1 by ImmunoGen.
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