title: Förster Resonance Energy Transfer-Based Self-Assembled Nanoprobe for Rapid and Sensitive Detection of Postoperative Pancreatic Fistula.
abstract: Postoperative pancreatic fistula (POPF) is the most serious and challenging complication following gastroenterological surgery. Activated pancreatic juice leaking from the organ remnant contains proteases that attack the surrounding tissue, potentially leading to severe inflammation, tissue necrosis, and fistula formation. However, it is difficult to observe pancreatic leakage during surgery and to evaluate the protease activity of leaked fluid at the patient's bedside. This report describes a protein nanocage-based protease ratiometric sensor comprising a pancreatic protease-sensitive small heat-shock protein (HSP) 16.5, which is a naturally occurring protein in Methanococcus jannaschii that forms a spherical structure by self-assembly of 24 subunits, and a chemically conjugated donor-acceptor dye pair for Förster resonance energy transfer (FRET). The HSP-FRET probe was constructed by subunit exchange of each dye-labeled engineered HSP, resulting in a spherical nanocage of approximately 10 nm in diameter, which exhibited very high stability against degradation in blood plasma and no remarkable toxicity in mice. The efficiency of FRET was found to depend on both the dye orientation and the acceptor/donor ratio. Pancreatic proteases, including trypsin, α-chymotrypsin, and elastase, were quantitatively analyzed by fluorescence recovery with high specificity using the HSP-FRET nanoprobe. Furthermore, the HSP-FRET nanoprobe was sufficiently sensitive to detect POPF in the pancreatic juice of patients using only the naked eye within 10 min. Thus, this novel nanoprobe is proposed as an effective and convenient tool for the detection of POPF and the visualization of activated pancreatic juice during gastroenterological surgery.
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Nobuhito
authors: N
authors: Hamano
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Masaharu
authors: M
authors: Murata
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Takahito
authors: T
authors: Kawano
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Jing Shu
authors: JS
authors: Piao
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Sayoko
authors: S
authors: Narahara
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Ryosuke
authors: R
authors: Nakata
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Tomohiko
authors: T
authors: Akahoshi
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Tetsuo
authors: T
authors: Ikeda
authors: Center for Advanced Medical Innovation, ‡Department of Advanced Medical Initiatives, Faculty of Medical Sciences, and §Innovation Center for Medical Redox Navigation, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors: Makoto
authors: M
authors: Hashizume
mesh_terms: D000818:Animals; D002918:Chymotrypsin / Q000737:chemistry / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D041981:Gastrointestinal Tract / Q000000981:diagnostic imaging* / Q000473:pathology / Q000601:surgery; D013502:General Surgery / Q000379:methods; D050888:Heat-Shock Proteins, Small / Q000737:chemistry*; D006801:Humans; D007249:Inflammation / Q000000981:diagnostic imaging* / Q000473:pathology; D063747:Methanocaldococcus / Q000737:chemistry; D051379:Mice; D049329:Nanostructures / Q000737:chemistry; D010185:Pancreatic Fistula / Q000000981:diagnostic imaging* / Q000473:pathology; D011183:Postoperative Complications / Q000000981:diagnostic imaging* / Q000473:pathology; D045663:Quantum Dots / Q000737:chemistry
chemical_list: D050888:Heat-Shock Proteins, Small; D002918:Chymotrypsin; C117608:alpha-chymotrypsin
keywords: FRET; clinical applications; pancreatic fistulas; self-assembled protein nanoprobes; visualization of leaked pancreatic juice