Conjugation of Panitumumab-IRDye800CW was performed under cGMP conditions, as described previously [3 (link)]. Briefly, Panitumumab (Vectibix; Amgen, Thousand Oaks, California, United States) was concentrated, and pH adjusted by buffer exchange to a 10 mg/mL solution in 50 mmol/L potassium phosphate, pH 8.5. IRDye800CW (IRDye800CW-N-hydroxysuccinimide ester, LI-COR Biosciences, Lincoln, Nebraska, United States) was conjugated to Panitumumab for 2 hours at 20°C in the dark, at a molar ratio of 2.3:1. After filtration with desalting columns (Pierce Biotechnology, Rockford, Illinois, United States) to remove unconjugated dye and buffer exchange to PBS, pH 7, the final protein concentration was adjusted to 2 mg/ml. The product was sterilized by filtration and placed into single-use vials and stored at 4°C until used.
Panitumumab
Manufactured by LI COR
Sourced in United States
Panitumumab is a monoclonal antibody developed by LI-COR. It is designed to target and bind to the epidermal growth factor receptor (EGFR) protein. This product is intended for use in various research applications, but a detailed description of its intended use is not available.
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2 protocols using panitumumab
1
Covalent Conjugation of Panitumumab with IRDye800CW
2
Fluorescent Antibody Conjugates for Imaging
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Panitumumab, anti-human Epidermal Growth Factor Receptor (hEGFR) antibody, was obtained from Amgen (Thousand Oaks, CA, USA; RRID AB_2459650 ). Anti-mouse CD29 antibody (clone IM7; RRID AB_2687714 ) was purchased from Bio X Cell (West Lebanon, NH, USA). Panitumumab (1 mg, 6·7 nmol) or anti-mouse CD29 antibody (1 mg, 6·7 nmol) was incubated with five-fold molar excess of IR700 NHS ester (10 mM in DMSO; LI-COR Biosciences, Lincoln, NE, USA) in 100 mM Na2HPO4 solution (pH 8·5) for 1 h at room temperature. The mixture was purified with PD-10 columns containing Sephadex G25 resin (Cytiva, Marlborough, MA, USA). The resulting AbPCs were abbreviated as pan-IR700 or CD29-IR700, respectively. 3′-OMe-DiR was synthesised as described previously.16 (link)
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