F-18 LABELING OF MONOCLONAL-ANTIBODIES AND FRAGMENTS WITH PRESERVATION OF IMMUNOREACTIVITY
F-18 LABELING OF MONOCLONAL-ANTIBODIES AND FRAGMENTS WITH PRESERVATION OF IMMUNOREACTIVITY
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DOI：
10.1021/bc00007a008
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发表时间：
1991-01-01
影响因子：
4.7
通讯作者：
ZALUTSKY, MR
中科院分区：
文献类型：
--
作者：
GARG, PK;GARG, S;ZALUTSKY, MR
报道了一种用正电子放射性核素F-18标记蛋白质的新方法。 最初，4-[F-18]-氟苄胺由[F-18]氟化物水溶液以高产率分两步制备。 该产物与辛二酸二琥珀酰亚胺酯反应生成F-18酰化剂。 在60 min的合成时间内，4-[F-18]氟苄胺琥珀酰亚胺酯（[F-18]SFBS）的总产率（衰变校正至回旋轰击结束）约为30%。反应15 min后，30-45%（衰变校正）的[F-18]SFBS可偶联至完整抗体及其F（ab '）2和Fab片段。 偶联产率依赖于蛋白质浓度，但不依赖于反应时间。 [F-18]SFBS的HPLC纯化是获得最佳偶联效率和免疫反应性所必需的。 F-18标记的抗肌球蛋白抗体的F（ab '）2和Fab片段的免疫反应性分别为89 +/- 5%和75 +/-9%。 在正常小鼠中的生物分布研究表明，F-18标记的抗体片段与使用N-琥珀酰亚胺基3-[I-125]碘苯甲酸酯标记的I-125标记的抗体片段的体内行为相似。 这些结果表明，该方法可能是有用的标记单克隆抗体和其他蛋白质和肽与F-18。
A new method is reported for labeling proteins with the positron-emitting nuclide F-18. Initially, 4-[F-18]-fluorobenzylamine was prepared in two steps from aqueous [F-18]fluoride in high yield. The F-18 acylation agent was formed by reaction of this product with disuccinimidyl suberate. Overall yields for the 4-[F-18]fluorobenzylamine succinimidyl ester ([F-18]SFBS), decay corrected to the end of cyclotron bombardment, were about 30% in a synthesis time of 60 min. After a 15-min reaction, 30-45% (decay corrected) of the [F-18]SFBS could be coupled to intact antibodies and their F(ab')2 and Fab fragments. Coupling yields were dependent on protein concentration but not reaction time. HPLC purification of [F-18]SFBS was necessary to obtain optimal coupling efficiency and immunoreactivity. The immunoreactivities of F-18-labeled F(ab')2 and Fab fragments of an antimyosin antibody were 89 +/- 5% and 75 +/- 9%, respectively. Biodistribution studies in normal mice demonstrated similar in vivo behavior of F-18-labeled antibody fragments and those labeled with I-125 by using N-succinimidyl 3-[I-125]iodobenzoate. These results indicate that this method may be useful for labeling monoclonal antibodies and other proteins and peptides with F-18.