Labelling small quantities of monoclonal antibodies and their F(ab')2 fragments with technetium-99m.
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.
Tesic
M
M
Department of Nuclear Medicine, Ontario Cancer Institute/Princess Margaret Hospital, Toronto, Canada.
Sheldon
K M
KM
Ballinger
J R
JR
Boxen
I
I
D000911:Antibodies, Monoclonal; D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007096:Imidoesters; D018796:Immunoconjugates; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification; D007074:Immunoglobulin G; D007553:Isotope Labeling / Q000379:methods; D008297:Male; D009362:Neoplasm Metastasis; D010434:Pepsin A; D017430:Prostate-Specific Antigen / Q000276:immunology; D013667:Technetium
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D007096:Imidoesters; D018796:Immunoconjugates; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; C005905:methyl 4-mercaptobutyrimidate; D013667:Technetium; D017430:Prostate-Specific Antigen; D010434:Pepsin A