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Labelling small quantities of monoclonal antibodies and their F(ab')2 fragments with technetium-99m
M Tesic1, K M Sheldon, J R Ballinger
1Department of Nuclear Medicine, Ontario Cancer Institute/Princess Margaret Hospital, Toronto, Canada.
Addition of sulfhydryl groups with 2-iminothiolane (2-IT) is an important new method for labelling monoclonal antibodies (mAb) and fragments with 99mTc. F(ab')2 fragments were prepared by digestion of 1B7.11 and BCD-F9 with pepsin. Optimal conditions for labelling 20-100 micrograms mAb or F(ab')2 involved a 2000:1 molar ratio of 2-IT:protein in phosphate buffer pH 7.4 for 30 min followed by addition of 99mTc-pertechnetate and stannous glucoheptonate. Recovered yields were > 70% and radiochemical purities were > 90% with a total preparation time of < 90 min.
Addition of sulfhydryl groups with 2-iminothiolane (2-IT) is an important new method for labelling monoclonal antibodies (mAb) and fragments with 99mTc. F(ab')2 fragments were prepared by digestion of 1B7.11 and BCD-F9 with pepsin. Optimal conditions for labelling 20-100 micrograms mAb or F(ab')2 involved a 2000:1 molar ratio of 2-IT:protein in phosphate buffer pH 7.4 for 30 min followed by addition of 99mTc-pertechnetate and stannous glucoheptonate. Recovered yields were > 70% and radiochemical purities were > 90% with a total preparation time of < 90 min.