1H NMR analysis was carried out on a AVANCE III HD nuclear magnetic resonance spectrometer at 400 MHz (Bruker, Germany). The molecular weight of AD was analyzed by mass spectrometry (Agilent, 6510 Q-TOF LC/MS, USA). The size, size distribution, and zeta potential of the nanoparticles in aqueous solution were determined by a Zetasizer Nano ZS instrument (Malvern, England) at 25°C. The nanoparticle morphology was visualized using Tecnai G2 20STwin transmission electron microscope (FEI, USA).
6510 q tof lc ms
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 6510 Q-TOF LC/MS is a high-resolution quadrupole time-of-flight mass spectrometer. It is designed for accurate mass measurement and advanced qualitative and quantitative analysis of complex samples.
Automatically generated - may contain errors
Lab products found in correlation
500 mhz nmr, by Agilent Technologies (2 mentions)
Tecnai g2 20 s twin, by Thermo Fisher Scientific (1 mentions)
Avance 3 hd, by Bruker (1 mentions)
Trypsin gold, by Promega (1 mentions)
Merck silica gel 60 f254, by Merck Group (1 mentions)
500 mhz nmr spectrometer, by Agilent Technologies (1 mentions)
Alanine, by Merck Group (1 mentions)
Phenylalanine, by Merck Group (1 mentions)
7 protocols using 6510 q tof lc ms
1
Nanoparticle Characterization by Advanced Analytical Techniques
2
Secretome Analysis of WT and Pvib-1 Strains
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Equal volume of culture supernatants of WT and Pvib-1 strains was subjected to SDS-PAGE and secretome proteins identified as described in [86] . In-gel trypsin-digestion was performed according to manufacture protocol (Promega, Trypsin Gold). Digested peptides were separated using ProtID-Chip-43 (II) and analyzed using the Agilent 6510 Q-TOF LC/MS as in [9] (link).
3
Drug-Antibody Ratio Determination by Mass Spectrometry
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DAR was determined using QTOF mass spectrometry (Agilent Technology 6510 QTOF LC/MS). Conjugated antibody (0.6 µg, 40 pmol) in 20 µL PBS was reduced with a 30 molar excess of TCEP for 2 hours at 37°C. The DAR of CC49‐VS‐MMAE was calculated as 2.27. The DAR of CC49‐Br‐MMAE was calculated as 10.0. The DAR of Dara‐Br‐MMAE was calculated as 10.0. The DAR of CC49‐mal‐MMAE was calculated as 9.54.
4
Synthesis and Characterization of Trifluoromethyl Substituted Compounds
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U1 –4 were synthesised by the procedure in the literature [11 (link)]. 3-Chloro-5-(trifluoromethyl)benzoic acid, 4-(trifluoromethoxy)benzoic acid, 3,4-diethoxycyclobut-3-ene-1,2-dione and COMU were purchased from Fluorochem Limited (Hadfield, UK). The remaining reagents were purchased from Merck (Darmstadt, Germany). Reactions were monitored using TLC on Merck silica gel 60 F254 aluminium-backed plates. Reaction products were purified as required with dry column vacuum chromatography on silica gel using gradient elutions. NMR spectra were recorded using a Bruker 400 MHz and an Agilent 500 MHz NMR spectrometer, at operating frequencies of 400 and 500 MHz for 1H and 100 and 125 MHz for 13C spectra, respectively. Spectra were referenced internally to the residual solvent (CDCl3: 1H δ 7.26, 13C δ 77.16. DMSO-d6: 1H δ 2.50, 13C δ 39.52. CD3OD: 1H δ 3.31, 13C δ 49.00). Multiplicity was assigned as s (singlet), d (doublet), t (triplet), q (quartet), qu (quintet) and m (multiplet). High resolution mass spectra (HRMS) were recorded on an Agilent Technologies 6510 Q-TOF LCMS. The purity of all test compounds was confirmed to be >95% using absolute quantitative NMR spectroscopy (Supplementary Information, Table S1 ) [17 (link)].
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5
Radiolabeling of DOTA-Conjugated Antibodies
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Dara or control trastuzumab (Tras) antibodies were reacted with a 30 molar excess of the chelator DOTA-NHS ester as previously described (6). DOTA conjugation was confirmed by Q-TOF mass spectrometry (Agilent Technology 6510 Q-TOF LC/MS) as follows: 6 µg of antibody was reduced with 1 µL of 0.2 M Tris(2-carboxyethyl)phosphine (TCEP) for 2 h at 37 °C and then analyzed on an HPLC protein Agilent chip (Agilent Technologies, Santa Clara, CA). DOTA-conjugated antibody (50 µg) was incubated with 225Ac at a labeling ratio of 1.85 MBq/µg for 45 min at 43 °C and chased with 1 mM DTPA. Radiolabeling efficiencies determined by instant thin layer chromatography were between 89 and 100% for all reactions.
6
Synthesis and Characterization of Ester-Protected Amino Acids
7
Synthesis and Characterization of Monounsaturated Fatty Acids
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(Z)-hexadec-13-enoic acid and (Z)-octadec-15-enoic acid were prepared by our previously reported procedures 39, 40 . (E)-octadec-9-enoic acid was purchased from Chemsupply (Gillman, SA, Australia). (Z)-hexadec-9-enoic acid, (Z)-octadec-11-enoic acid, (E)-octadec-11-enoic acid, (Z)-octadec-6-enoic acid and all other reagents and anhydrous solvents were purchased from Sigma Aldrich (Castle Hill, NSW, Australia).
Non-commercially available MUFAs were synthesised as described in the supporting information. The purity of all test compounds (3 -20) was confirmed by elemental analysis carried out in the Campbell Microanalytical Laboratory at the Department of Chemistry, University of Otago. All values were within ±0.4% of the calculated values.
Dry Column Vacuum Chromatography (DCVC) was used to purify reaction products on silica gel with gradient elutions. TLC was performed on silica gel 60 F254 plates. 1 H and 13 C NMR spectra were recorded on an Agilent 500 MHz NMR. Spectra were referenced internally to residual solvent (CDCl3; 1 H 7.26, 13 C 77.10. d6-DMSO; 1 H 2.49, 13 C 39.52). High resolution mass spectra were recorded on Agilent 6510 Q-TOF LC/MS.
Non-commercially available MUFAs were synthesised as described in the supporting information. The purity of all test compounds (3 -20) was confirmed by elemental analysis carried out in the Campbell Microanalytical Laboratory at the Department of Chemistry, University of Otago. All values were within ±0.4% of the calculated values.
Dry Column Vacuum Chromatography (DCVC) was used to purify reaction products on silica gel with gradient elutions. TLC was performed on silica gel 60 F254 plates. 1 H and 13 C NMR spectra were recorded on an Agilent 500 MHz NMR. Spectra were referenced internally to residual solvent (CDCl3; 1 H 7.26, 13 C 77.10. d6-DMSO; 1 H 2.49, 13 C 39.52). High resolution mass spectra were recorded on Agilent 6510 Q-TOF LC/MS.
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