The gold nanoparticles coated with cetuximab induce apoptosis in cancer cells even under non-physiological conditions raw data
Description
This document presents the raw data regarding the effects of acidic and alkaline solutions, temperature and buffers on the colloidal stability of uncovered gold nanoparticles and functionalized gold nanoparticles with PEG and cetuximab, a monoclonal antibody. Our hypotesis is centered whether covered gold nanoparticles surface protects these nanoparticles of aggregation. Moreover, the nanoplatform cetuximab-gold nanoparticles mantains its EGFR targeting affinity independentlly of pH and temperature evaluated mimicking tumor microenvironment fashion. To validade this hypothesis a setup of charactterization was performed, mainly spectroscopy. The raw data available here can shed light on the challenges of collecting a such data, as well as can inspire other researdchs to choose more adequate characterization setups to help them to get answears from hypothesis related to colloidal stability of bare and functionalized gold nanoparticles.
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It is necessary carefully preparation of samples accoding to standard protocols and follow technical especifications of each equipement such as UHV, sterilization conditions, to observe the quality of buffers and solutions. We used ICP-OES parameters were used: nebulizer flow rate, 0.80 L·min⁻¹; radio frequency power, 1450 W; sample introduction rate, 1.50 mL·min⁻¹; flush time, 15 s; delay time, 20 s; read time, 10 s; and wash time, 60 s. All measurements were performed in triplicate, and results are reported as µg of Au per mL. Circular dichroism (CD) spectra of AuNPCet at the pH values of 2.9, 8.9 and 10.3 and at different temperatures, 5 oC, 36 oC and 40 oC were recorded in spectropolarimeter J-815 equipped with a PTC-423S Peltier controller for temperature AuNP, AuNPPEG and AuNPCet were titrated in a potentiometric titrator Metrohm Titrando 808 (Switzerland). 5 mL of each sample was used in each titration. For the acid curve, a HCl 0.01 mol L -1 was used as titrant. For the basic curve, a NaOH 0.01 mol L -1 was used as titrant under stirring. Gold nanoparticles were labelled with secondary antibody Alexa Fluor® 488 IgG H&L, dilution 1:400µL. (Molecular Probes, USA) for 30 minutes at room temperature and protected from light. Fluorescence was acquired by Cytation™ 5 cell imaging multimode reader equipment (BioTek, USA) according to excitation λ 495nm and emission λ 519nm.Cells were seeded in individual optical 35mm glass bottom petri-dishes (Nunc™, USA) 24h before experiments. Cells were exposed to cetuximab and AuNPCet for 60 minutes at 4oC. Another experimental setup A431 cells were treated with AuNPCet acid and alkaline pH adjusted and incubated for 60 minutes at 4oC. Next, the samples were labelled with Alexa Fluor® 488 IgG H&L and Hoechst 33342 for 30 minutes at dark. Images were acquired in the LSM 800 laser scanning Airyscan confocal microscope .TGA measurements were performed with a simultaneous DTA-TG apparatus DTG-60H from Shimadzu. (Brazil) Approximately 5mg of the materials cetuximab (Erbitux®, Merck, Germany), sodium citrate (Sigma-Aldrich, New York, USA) and AuNPCet (lyophilized) were used. Samples were placed in 50 µL alumina pans and then heated at a rate of 5 °C min-1 to 1000 °C. The analysis was performed in an inert atmosphere, using a nitrogen gas flow of 50 mL min-1.. XPS measurements were acquired with an ESCALAB spectrometer from Thermo Fisher (Germany) using monochromatized Al-K α (1486.68eV) radiation at a base pressure of 2 × 10 -10 mbar. Survey spectra were collected with pass energy of 50eV, while high-resolution spectra of the C1s, O1s, Au4f and N1s lines were taken at E pass = 20eV.
Institutions
- Universidade Federal de Minas GeraisMinas Gerais, Belo Horizonte