A drug-linker molecule
VcMMAE is a drug-linker molecule that has been used in the synthesis of antibody-drug conjugates (ADCs).1,2 It contains a maleimidocaproyl (Mc) group that can be conjugated to an antibody, a protease-cleavable peptide linker, and the antimitotic and anticancer agent monomethyl auristatin E .
1.Chuprakov, S., Ogunkoya, A.O., Barfield, R.M., et al.Tandem-cleavage linkers improve the in vivo stability and tolerability of antibody-drug conjugatesBioconjug. Chem.32(4)746-754(2021) 2.Dimou, M., Papageorgiou, S.G.S., N., Katodritou, E., et al.Real-life experience with the combination of polatuzumab vedotin, rituximab, and bendamustine in aggressive B-cell lymphomasHematol. Oncol.39(3)336-348(2021)
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Cell experiment: |
Monomethyl auristatin E (MMAE, 5 nM) and ionizing radiation (IR) treated cells are harvested and lysed in RIPA buffer with protease and phosphatase inhibitors. 30μg of lysate undergo electrophoresis using 4-12% Bis-Tris gels, transferred to PVDF membranes and incubated with indicated primary antibodies. Blots are developed by ECL. |
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Animal experiment: |
6-8 week old female athymic nu/nu mice are injected subcutaneously into thighs with 5×106 HCT-116 or PANC-1 cells in a 1:1 Matrigel and PBS solution. Mice are treated with IR or intravenous (IV) injection of ACPP-cRGD-MMAE (6 nmoles/day, 18 nmoles total, i.v.), tumor tissue is harvested, formalin fixed and paraffin embedded followed by staining with indicated antibodies. The primary antibody is used at a 1:250 dilution and is visualized using DAB as a chromagen with the UltraMap system. |
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参考文献: [1]. Okeley, et al. Intracellular Activation of SGN-35, a Potent Anti-CD30 Antibody-Drug Conjugate. Clinical Cancer Research (2010), 16(3), 888-897. |
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