title: Biological production of an integrin αvβ3 targeting imaging probe and functional verification.
abstract: The aim of the present study is to establish a bacterial clone capable of secreting an integrin αvβ3 targeting probe with bioluminescent and fluorescent activities, and to verify its specific targeting and optical activities using molecular imaging. A bacterial vector expressing a fusion of secretory Gaussia luciferase (sGluc), mCherry, and RGD (sGluc-mCherry-RGDX3; GCR), and a control vector expressing a fusion of secretory Gaussia luciferase and mCherry (sGluc-mCherry; GC) were constructed. The GCR and GC proteins were expressed in E. coli and secreted into the growth medium, which showed an approximately 10-fold higher luciferase activity than the bacterial lysate. Successful purification of GCR and GC was achieved using the 6X His-tag method. The GCR protein bound with higher affinity to U87MG cells than CHO cells in confocal microscopy and IVIS imaging, and also showed a high affinity for integrin αvβ3 expressing tumor xenografts in an in vivo animal model. An E. coli clone was established to secrete an integrin αvβ3 targeting imaging probe with bioluminescent and fluorescent activities. The probe was produced feasibly and at low cost, and has shown to be useful for the assessment of angiogenesis in vitro and in vivo.
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Mi-Hye
authors: MH
authors: Hwang
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Jung Eun
authors: JE
authors: Kim
authors: Department of Biochemistry and Cell Biology, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea.
authors: Sang-Yeob
authors: SY
authors: Kim
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Senthilkumar
authors: S
authors: Kalimuthu
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Shin Young
authors: SY
authors: Jeong
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Sang-Woo
authors: SW
authors: Lee
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Jaetae
authors: J
authors: Lee
authors: Department of Nuclear Medicine, Kyungpook National University School of Medicine, 50 Samduk-dong 2-ga, Jung Gu, Daegu 700-721, Republic of Korea.
authors: Byeong-Cheol
authors: BC
authors: Ahn
mesh_terms: D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D057054:Molecular Imaging; D015335:Molecular Probes / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D015203:Reproducibility of Results; D023041:Xenograft Model Antitumor Assays
chemical_list: D039302:Integrin alphaVbeta3; D015335:Molecular Probes; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C047981:arginyl-glycyl-aspartic acid; D008156:Luciferases
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