MBP-annexin V radiolabeled directly with iodine-124 can be used to image apoptosis in vivo using PET.
A noninvasive method of measuring programmed cell death in the tumors of cancer patients using positron-emission tomography (PET) would provide valuable information regarding their response to therapeutic intervention. Our strategy is to radiolabel annexin V, a protein that binds to phosphatidylserine moieties that are translocated to the external leaflet of plasma membranes during apoptosis. We developed a phosphatidylserine-ELISA capable of distinguishing wild type from point mutant annexin V that is known to have a lower phosphatidylserine binding affinity. A maltose-binding protein/annexin V chimera was synthesized and purified with high yield using amylose resin. We showed that it bound to phosphatidylserine in the ELISA as well as to that exposed on apoptotic Jurkat cells; therefore, it was used in the development of a method for radiolabeling annexin V using iodine radionuclides. MBP-annexin V retained its phosphatidylserine binding properties on direct iodination, but at high levels of oxidizing agents (iodogen and chloramine T), its specificity for phosphatidylserine was compromised. (124)I-MBP-annexin V was successfully used to image Fas-mediated hepatic cell death in BDF-1 mice using PET. In conclusion, we have shown that MBP-annexin V and the phosphatidylserine ELISA are useful tools for the development of methods for radiolabeling annexin V for PET imaging.
Dekker
Bronwen
B
Cancer Research UK/UMIST, Department of Radiochemical Targeting and Imaging, Paterson Institute for Cancer Research, M20 4BX Manchester, UK.
Keen
Heather
H
Lyons
Stephen
S
Disley
Lynn
L
Hastings
David
D
Reader
Andrew
A
Ottewell
Penny
P
Watson
Alastair
A
Zweit
Jamal
J
D000818:Animals; D017304:Annexin A5 / Q000737:chemistry / Q000493:pharmacokinetics*; D017209:Apoptosis; D002352:Carrier Proteins / Q000737:chemistry / Q000493:pharmacokinetics; D022781:Hepatocytes / Q000000981:diagnostic imaging* / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes / Q000737:chemistry / Q000493:pharmacokinetics; D019169:Jurkat Cells; D008099:Liver / Q000000981:diagnostic imaging* / Q000378:metabolism; D008297:Male; D058619:Maltose-Binding Proteins; D008657:Metabolic Clearance Rate; D051379:Mice; D009928:Organ Specificity; D049268:Positron-Emission Tomography / Q000379:methods*; D019275:Radiopharmaceuticals / Q000138:chemical synthesis / Q000493:pharmacokinetics; D014018:Tissue Distribution
D017304:Annexin A5; D002352:Carrier Proteins; D007457:Iodine Radioisotopes; D058619:Maltose-Binding Proteins; D019275:Radiopharmaceuticals