Methods to avoid adverse effect of circulating antigen on biodistribution of 125I-labeled antiTac dsFv: preinjection of intact antibody versus clearance of antigen with adivin-biotin system.
The presence of circulating antigen may adversely affect the biodistribution of a radiolabeled antibody. The alpha subunit of the interleukin-2 receptor (IL-2Ralpha) is a cell-surface receptor that is overexpressed in various hematologic malignancies and in benign disorders. This receptor is cleaved from the cell surface and can be found in high concentrations in serum. Radiolabeled antiTac antibodies are being evaluated to target this receptor. Previous studies have shown that circulating soluble IL-2Ralpha (slL-2Ralpha) adversely affected the biodistribution of radiolabeled antiTac disulfide-stabilized (ds)Fv. In this study, we compared blocking and clearing sIL-2Ralpha to see which better minimized its interference with the biodistribution of radiolabeled antiTac dsFv.


METHODS
Two models of sIL-2Ralpha were used: one consisted of mice given intravenous sIL-2Ralpha and the other consisted of mice bearing SP2/Tac tumor xenografts (IL-2Ralpha positive), which shed sIL-2Ralpha. We biotinylated humanized antiTac monoclonal antibody (bt-HuTac) and radiolabeled it with 125I. We then compared its biodistribution with that of humanized antiTac monoclonal antibody IgG (HuTac). We examined the biodistribution of an injected dose of 125I-labeled antiTac dsFv after a preinjection of HuTac to block the sIL-2Ralpha epitope and after a preinjection of bt-HuTac, followed by an avidin chase.


RESULTS
The 125I-labeled bt-HuTac cleared from the serum at a rate similar to that of HuTac. The avidin chase effectively cleared >92% of circulating 125I-labeled bt-HuTac within 20 min and was also effective in clearing sIL-2Ralpha. In comparison, HuTac prolonged the retention of 125I-labeled sIL-2Ralpha in the circulation, and the avidin chase decreased 125I-labeled sIL-2Ralpha to <18% of control. Although the two-step antigen-clearing system effectively cleared the antigen from the circulation and improved the biodistribution of 125I-labeled dsFv, the HuTac preinjection method had a similar but longer lasting beneficial effect on 125I-labeled dsFv biodistribution.


CONCLUSIONS
Preinjection of either HuTac or bt-HuTac with avidin chase improved the biodistribution of subsequently administered 125I-labeled antiTac dsFv by preventing the dsFv from binding to the sIL-2Ralpha, but the HuTac blocking method is simpler and longer lasting.
Kobayashi
H
H
Department of Nuclear Medicine, Warren G. Magnuson Clinical Center, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-1180, USA.
Sun
B F
BF
Yoo
T M
TM
Le
N
N
Kim
M K
MK
Paik
C H
CH
Pastan
I
I
Waldmann
T A
TA
Carrasquillo
J A
JA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000493:pharmacokinetics*; D001360:Avidin / Q000493:pharmacokinetics*; D019921:Biotinylation; D002851:Chromatography, High Pressure Liquid; D007128:Immunoglobulin Fragments / Q000378:metabolism*; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D007457:Iodine Radioisotopes / Q000493:pharmacokinetics*; D051379:Mice; D008819:Mice, Nude; D015375:Receptors, Interleukin-2 / Q000276:immunology*; D011994:Recombinant Proteins / Q000378:metabolism; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D007457:Iodine Radioisotopes; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D001360:Avidin