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Optimal Quality ¹³¹I-Monoclonal Antibodies on High-Dose Labeling in a Large Reaction Volume and Temporarily Coating the Antibody with IODO-GEN

Radionuclide Center Vrije Universiteit and Departments of Obstetrics and Gynecology, Otolaryngology, and Nuclear Medicine/PET Center, University Hospital Vrije Universiteit, Amsterdam, The Netherlands

View larger version (12K): [in a new window]   FIGURE 1. Phosphor imager profile of SDS-PAGE gel of 131I-c-MOv18, high-dose–labeled with conventional IODO-GEN–coated vial method. Quantification report: 1, 1.1%; 2, 8.6%; 3, 70.4%; 4, 8.7%; 5, 1.2%; 6, 1.1%; 7, 8.9%. Conditions: 4.4 GBq 131I, 5 mg c-MOv18, 500 µg IODO-GEN, 2-mL reaction volume, 10-min reaction time.

View larger version (11K): [in a new window]   FIGURE 2. HPLC profile of 131I quality of aged 131I solution in blackened delivery vial (3.7 GBq in 500 µL) 4 d after production (A) and after adjustment of pH, transfer to reaction vial, and addition of ascorbic acid (B). Percentages: (A) 13% 131IO3- (retention time [Rt], 1.7 min), 79% 131I- (Rt, 5.0 min), 8% unidentified 131I components. (B) 100% 131I- (Rt, 5.0 min).

View larger version (21K): [in a new window]   FIGURE 3. HPLC visualization of temporary coating of antibody with IODO-GEN and subsequent removal of coating by ascorbic acid through simultaneous UV detection at 315 nm, second absorption maximum of IODO-GEN (UV 2), and, at 280 nm, absorption maximum of MAb (UV 1). Shown are native MAb (A), coated MAb (B), and MAb with coating removed (C). Conditions are described in text. Large peak at 40 min (C) is from ascorbic acid; minor peaks (at 16 and 39 min) in chromatograms (A and B) at 315 nm are solvent peaks.

View larger version (12K): [in a new window]   FIGURE 4. Phosphor imager profile of SDS-PAGE gel of 131I-c-MOv18, 1 h (A) and 24 h (B) after formulation. Radioactivity concentration: 0.37 GBq/mL. Quantification report: (A) 1, 0.4%; 2, 98.3%; 3, 1.3%. (B) 1, 1.5%; 2, 80.1%; 3, 10.9%; 4, 2.7%; 5, 1.4%; 6, 3.3%. Conditions (IODO-GEN–coated MAb method): 4.3 GBq 131I, 5 mg c-MOv18, 35 µg IODO-GEN, 6-mL reaction volume, 3-min reaction time.

View larger version (10K): [in a new window]   FIGURE 5. Immunoreactivity assay of radioiodinated c-MOv18 MAbs used for biodistribution analysis: 125I-c-MOv18 (); 131I-c-MOv18, high dose (); 131I-c-MOv18, low dose (•). T/C values (T = total radioactivity; C = activity specifically bound to cells) are function of inverse of cell concentration.