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Maleimido mono amide dota

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Maleimido-mono-amide-DOTA is a bifunctional metal chelator that can be used to label biomolecules with metal ions for various applications. It contains a maleimide group for covalent conjugation to biomolecules and a DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) chelating group for complexation of metal ions.

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Lab products found in correlation

Inveon pet ct system, by Siemens (1 mentions) Micropet focus 220, by Siemens (1 mentions) Ethylene diamine tetraacetic acid (edta), by STEMCELL (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Sephadex g 25, by GE Healthcare (1 mentions) Nanodrop lite, by Thermo Fisher Scientific (1 mentions) Cetuximab erbitux, by Merck Group (1 mentions)

6 protocols using maleimido mono amide dota

1

PET Imaging of CCR2 and CCR5 Receptors

Cited 2 times
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The CCR2 (ECL1i: LGTFLKC) and CCR5 (DAPTA: D-A1STTTNYT) targeting peptides were synthesized from D-form amino acids by CPC Scientific (Sunnyvale, CA). Maleimido-mono-amide-DOTA was purchased from Macrocyclics, Inc (Dallas, TX). DOTA-ECL1i and DOTA-DAPTA was synthesized as reported (Heo et al., 2019 ; Liu et al., 2017 (link); Luehmann et al., 2014 (link)). Probes were radiolabeling with 64Copper (64Cu). The radiolabeled compound was analyzed using radio-HPLC to ensure more than 95% radiochemical purity prior to animal studies. Mice were anesthetized with isoflurane and injected with 3.7 MBq of 64Cu-DOTA-ECL1i or 3.7 MBq 64Cu-DOTA-DAPTA in 100 μL of saline via the tail vein. Small animal PET scans (0 to 60 min dynamic scan) were performed on either microPET Focus 220 (Siemens, Malvern, PA) or Inveon PET/CT system (Siemens, Malvern, PA). The microPET images were corrected for attenuation, scatter, normalization, and camera dead time and co-registered with microCT images. All of the PET scanners were cross-calibrated periodically. The microPET images were reconstructed with the maximum a posteriori (MAP) algorithm and analyzed by Inveon Research Workplace. The uptake was calculated as the percent injected dose per gram (%ID/gram) of tissue in three-dimensional regions of interest (ROIs) without the correction for partial volume effect.
Wong N.R., Mohan J., Kopecky B.J., Guo S., Du L., Leid J., Feng G., Lokshina I., Dmytrenko O., Luehmann H., Bajpai G., Ewald L., Bell L., Patel N., Bredemeyer A., Weinheimer C.J., Nigro J.M., Kovacs A., Morimoto S., Bayguinov P.O., Fisher M.R., Stump W.T., Greenberg M., Fitzpatrick J.A., Epelman S., Kreisel D., Sah R., Liu Y., Hu H, & Lavine K.J. (2021). Resident Cardiac Macrophages Mediate Adaptive Myocardial Remodeling. Immunity, 54(9), 2072-2088.e7.
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2

Mass Cytometry Cell Barcoding Protocol

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To minimize inter-sample staining variation, we applied mass-tag barcoding to fixed cells (Zunder et al., 2015 (link)). A 126-well barcoding scheme composed of unique combinations of four out of nine barcoding metals was used for this study; metals included palladium (105Pd, 106Pd, 108Pd, 110Pd, Fluidigm) conjugated to bromoacetamidobenzyl-EDTA (Dojindo) as well as indium (113In and 115In, Fluidigm), yttrium, rhodium, and bismuth (89Y, 103Rh, 209Bi, Sigma Aldrich) conjugated to maleimido-mono-amide-DOTA (Macrocyclics). The concentrations were adjusted to 20 nM (209Bi), 100 nM (105-110Pd, 115In, 89Y), 200 nM (113In), or 2 μM (103Rh). Cells were randomly distributed across two 96-well plates, and about 0.3 million cells per well were barcoded using a transient partial permeabilization protocol. Cells were washed once with 0.03% saponin in PBS (Sigma Aldrich) prior to incubation in 200 μl barcoding reagent for 30 min at room temperature. Cells were then washed four times with cell staining medium (CSM, PBS with 0.3% saponin, 0,5% bovine serum albumin (Sigma Aldrich) supplemented with 2 mM EDTA (StemCell Technologies, Inc.) and pooled for antibody staining. Two 126-well barcoding plates, with a set of standard samples on each plate, were used for antibody staining with the tumor cell-centric and the immune cell-centric panels (Tables S3 and S4, respectively).
Wagner J., Rapsomaniki M.A., Chevrier S., Anzeneder T., Langwieder C., Dykgers A., Rees M., Ramaswamy A., Muenst S., Soysal S.D., Jacobs A., Windhager J., Silina K., van den Broek M., Dedes K.J., Rodríguez Martínez M., Weber W.P, & Bodenmiller B. (2019). A Single-Cell Atlas of the Tumor and Immune Ecosystem of Human Breast Cancer. Cell, 177(5), 1330-1345.e18.
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3

Single-Cell Palladium Barcoding Protocol

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Reagents for barcoding were prepared as previously described.15 (link) Two molar equivalents of maleimido-mono-amide-DOTA (Macrocyclics, Inc., Dallas, TX) were added to palladium 102, 104,105, 106, 108, and 110 solutions in 20 mM ammonium acetate, pH 6.0. Solutions were immediately lyophilized, and solids were dissolved in dimethyl sulfoxide (DMSO) to 10 mM for storage at −20 °C. Each well of a barcoding plate contained a distinct combination of three palladium isotopes at 200 nM in DMSO.
After thawing and red blood cell lysis in a hypotonic buffer, cells were barcoded as previously described.16 (link) Briefly, cells were transferred into a deep-well block and washed once with cell staining media (CSM, PBS with 0.5% BSA, 0.02% NaN3), once with PBS, and once with 0.02% saponin in PBS. The barcoding plate was thawed, and each well of barcode reagent was diluted in 1 mL 0.02% saponin in PBS. Diluted barcode reagent was transferred to cells, and samples were incubated at room temperature for 15 minutes, washed twice with CSM, and then pooled for staining.
Fragiadakis G.K., Gaudillière B., Ganio E.A., Aghaeepour N., Tingle M., Nolan G.P, & Angst M.S. (2015). Patient-specific Immune States before Surgery are Strong Correlates of Surgical Recovery. Anesthesiology, 123(6), 1241-1255.
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4

Palladium Isotope Barcoding Protocol

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Reagents were prepared according to the procedure described in Bodenmiller et al. (25 ). Two molar equivalents of maleimido-mono-amide-DOTA (Macrocyclics, Inc.) were added to palladium 102, 104, 105, 106, 108, 110, each contained in 20 mM ammonium acetate at pH 6.0. Solutions were immediately lyophilized, and solids were dissolved in dimethyl sulfoxide (DMSO) to 10 mM for storage at −20 °C. Each well of a barcoding plate contained a unique combination of three palladium isotopes at 200 nM in DMSO.
Gaudilliere B., Fragiadakis G.K., Bruggner R.V., Nicolau M., Finck R., Tingle M., Silva J., Ganio E.A., Yeh C.G., Maloney W.J., Huddleston J.I., Goodman S.B., Davis M.M., Bendall S.C., Fantl W.J., Angst M.S, & Nolan G.P. (2014). Coordinated Surgical Immune Signatures Contain Correlates of Clinical Recovery. Science translational medicine, 6(255), 255ra131.
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5

Radiolabeling and Characterization of ECL1i

Cited 1 time
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The ECL1i peptide (LGTFLKC) was synthesized from D-form amino acids by CPC Scientific (Sunnyvale, CA). DOTA-ECL1i was prepared by conjugating 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) that was modified as maleimido-mono-amide-DOTA (1.573 mg, 0.2 μmol; Macrocyclics, Dallas, TX) to the cysteine residue of ECL1i, using established methods (22 (link)). The crude conjugate was purified by high performance liquid chromatography (HPLC) to reach 99% chemical purity and verified by mass spectrometry. Copper-64 (64Cu) was selected as an initial radiolabel for ECL1i based on a high specific activity that enabled trace amount administration and provided a decisive PET signal if present, straightforward radiochemistry through the conjugation of the DOTA chelator on the peptide, on-site availability, and prior experience with this radionuclide (22 (link)). The DOTA-ECL1i conjugate was radiolabeled with 64CuCl2 as described (22 (link)). Copper-64-DOTA-ECL1i (64Cu-DOTA-ECL1i) was tested for stability by incubation in mouse serum at 37 °C for 1 h and in vivo, in blood and lung 1 h post injection (n=3), by radio-HPLC analysis as described in the Supplement. Maleimide-modified Dylight 550 (ThermoFisher Scientific, Waltham, MA) was conjugated to ECL1i on the cysteine residue following the same protocol for DOTA conjugation and purified by HPLC.
Liu Y., Gunsten S.P., Sultan D.H., Luehmann H.P., Zhao Y., Blackwell T.S., Bollermann-Nowlis Z., Pan J.H., Byers D.E., Atkinson J.J., Kreisel D., Holtzman M.J., Gropler R.J., Combadiere C, & Brody S.L. (2017). PET-based Imaging of Chemokine Receptor 2 in Experimental and Disease-related Lung Inflammation. Radiology, 283(3), 758-768.
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6

Characterization of Antibody-Based Therapeutics

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Rituximab (Rituxan; lot: R1102AB)
and trastuzumab (Herceptin; lot: 19B020E) were available from Chugai
Pharmaceutical Co. Ltd. (Chuo-ku, Tokyo, Japan). Cetuximab (Erbitux;
lot: ERBA012) and mogamulizumab (Poteligeo; lot: 19303P) were purchased
from Merck Biopharma (Meguro-ku, Tokyo, Japan) and Kyowa Hakko Kirin
Co. Ltd. (Chiyoda-ku, Tokyo, Japan), respectively. Payload (dibenzocyclooctyne-PEG4-biotin) was purchased from Sigma-Aldrich (St. Louis, MO,
USA). Bifunctional chelating agents, p-SCN-Bn-DOTA
and maleimido-mono-amide-DOTA, were purchased from Macrocyclics (Dallas,
TX, USA). Sialylglycan and 2-chloro-1,3-dimethyl-1H-benzimidazol-3-ium chloride for preparing oxazoline were from Fushimi
Pharmaceutical Co., Ltd. (Marugame, Kagawa).
A TSKgel FcR-IIIA-NPR
(4.6 mm I.D. × 7.5 cm) column from TOSOH (Minato-ku, Tokyo, Japan)
was used for the analysis of all samples. Protein A column chromatography
was available from TOSOH (Minato-ku, Tokyo, Japan). HPLC (Prominence,
SHIMADZU, Kyoto, Japan) was used for analyses and purification. The
antibody concentration was measured by a NanoDrop Lite (Thermo Fisher
Scientific, Waltham, MA, USA). The 1H-spectra of oxazolines
were measured on a JEOL JMN-ECZL spectrometer (400 MHz) at ambient
temperature in D2O. Sephadex G-25 was purchased from GE
Healthcare (Chicago, IL, USA) for purification of glycan oxazoline.
Hiranyakorn M., Iwamoto S., Hoshinoo A., Tsumura R., Takashima H., Yasunaga M, & Manabe S. (2023). Chromatographic Analysis of the N-Glycan Profile on Therapeutic Antibodies Using FcγRIIIa Affinity Column Chromatography. ACS Omega, 8(18), 16513-16518.
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