Conjugation of cetuximab-IRDye800 was performed under cGMP conditions, as previously described.(19 (link)) Briefly, cetuximab® (ImClone LLC, Eli Lilly and Company, Branchburg, NJ) was concentrated and pH adjusted by buffer exchange to a 10mg/ml solution in 50mM potassium phosphate, pH 8.5. IRDye800CW NHS ester (LI–COR Biosciences, Lincoln NE) was conjugated to cetuximab for 2hr at 20°C in the dark, at a molar ratio of 2.3:1.
Protocol detail
Cetuximab-IRDye800 Conjugation Protocol
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Corresponding Organization :
Other organizations : University of Alabama at Birmingham, University Medical Center Groningen, University of Groningen
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Variable analysis
independent variables
- Molar ratio of IRDye800CW NHS ester to cetuximab (2.3:1)
- Degree of conjugation of IRDye800CW to cetuximab
- Concentration of cetuximab (10 mg/ml)
- PH of the reaction (8.5)
- Temperature of the reaction (20°C)
- Duration of the reaction (2 hours)
- Reaction performed under cGMP conditions
- None explicitly mentioned
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