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I-124 Labeled recombinant human Annexin V produced by E. coli for apoptosis image using small animal PET

¹ Division of Nuclear Medicine and RI Application, KIRAMS, Seoul, South Korea

1293

Objectives: Annexin V labeled with radioisotope and optical probe has been used to detect apoptosis. To evaluate annexin V as a multimodal apoptosis image agent, large-scale preparation of Annexin V (AV) is prerequisite. The aim of this study is to produce and purify recombinant human Annexin V (rh-AV) in E. coli system and to evaluated radiolabeled rh-AV in vitro and in vivo apoptosis model system.

Methods: Annexin V cDNA was obtained from human placenta and rh-AV cloned in E. coli pET 28a (+) vector, expressed and purified by His-Tag and T7-Tag resins. In camptothecin induced Jurkat T cell apoptosis model, AV-PI flow cytometry analysis and in vitro binding assay of I-124 rh-AV were performed and compared. Biodistributions and small animal PET images of I-124 AV and rh-AV were evaluated and compared in normal and Fas-madiated hepatic apoptosis Balb/c mice models.

Results: Through two step purification process, rh-AV showed 35 Kd single band in SDS-PAGE. As camptothecin concentration increasing, AV-FITC positive % increased in flow cytometry analysis and uptake of I-124 rh-AV also increased. AV-FITC positive % was correlated with and uptake of I-124 rh-AV (R2=0.99). I-124 rh-AV showed faster blood clearance and lower non specific uptake (liver and kidney) than I-124 AV. In Fas-mediated hepatic apoptosis model, I-124 rh-AV (3.8 %ID/g) was more selectively localized in liver region of PET image than I-124 AV (3.49 %ID/g).

Conclusions: Recombinant Human annexin V was produced by E. coli system and purified using two step affinity chromatography. Radiolabeled rh-AV was useful for the evaluation of apoptosis in vitro and in vivo model. Recombinant human annexin V could be used as apoptosis imaging agent with various radiolabel and optical probe.

This Article Services Email this article to a friend Similar articles in this journal Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Jung, J. Articles by Lim, S. PubMed Articles by Jung, J. Articles by Lim, S.