title: Preparation of magnetic resonance probes using one-pot method for detection of hepatocellular carcinoma.
abstract: OBJECTIVE
  To prepare the specific magnetic resonance (MR) probes for detection of hepatocellular carcinoma (HCC) using one-pot method.
  
  
  METHODS
  The carboxylated dextran-coated nanoparticles were conjugated with anti-α-fetoprotein (anti-AFP) or anti-glypican 3 (anti-GPC3) antibodies through 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS)-mediated reaction to synthesize the probes. The physical and chemical properties of the probes were determined by transmission electron microscopy (TEM) and dynamic light scattering, and the relaxivity was compared to uncombined ultrasmall superparamagnetic iron oxide nanoparticles (USPIONs) using a 1.5T clinical MR scanner. The binding efficiency of the antibodies to nanoparticles was measured with an ultraviolet-visible spectrophotometer. In addition, the probes were incubated with targetable cells in vitro.
  
  
  RESULTS
  The superparamagnetic MR probes (anti-GPC3-USPION probe and anti-AFP-USPION probe) were synthesized using one-pot method. Their mean hydrodynamic diameter was 47 nm with a broader slight size distribution. The coupling efficiency of carboxylated dextran-coated ultrasmall superparamagnetic iron oxide (USPIO) with anti-GPC3 or anti-AFP antibody was 15.9% and 88.8%, respectively. Each of the USPIO nanoparticles may bind 3 GPC3 antibodies or 12 AFP antibodies. The statistical analysis showed no significance (P > 0.05) in shortening the T1 and T2 values when comparing the USPIO-AFP or USPIO-GPC3 to USPIO. Analysis of TEM images revealed that anti-GPC3-USPION probes and anti-AFP-USPION probes could specifically enter into the HepG2 cell by combining with the GPC3 receptors or AFP receptors, whereas the HepG2 cell sample incubated with USPIONs showed no or few nanoparticles in the cytoplasm.
  
  
  CONCLUSIONS
  The synthesized probes using one-pot method can be used for in vitro experimental study and have potential clinical application in MR imaging for detection of hepatocellular carcinomas.
authors.affiliation: You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
authors.forename: You-Wei
authors.initials: YW
authors.lastname: Li
authors.affiliation: You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
authors.forename: Zheng-Guang
authors.initials: ZG
authors.lastname: Chen
authors.affiliation: You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
authors.forename: Zhou-She
authors.initials: ZS
authors.lastname: Zhao
authors.affiliation: You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
authors.forename: Hong-Li
authors.initials: HL
authors.lastname: Li
authors.affiliation: You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
authors.forename: Ji-Chen
authors.initials: JC
authors.lastname: Wang
authors.affiliation: You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
authors.forename: Zong-Ming
authors.initials: ZM
authors.lastname: Zhang
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D003287:Contrast Media / Q000138:chemical synthesis* / Q000378:metabolism; D003911:Dextrans / Q000138:chemical synthesis / Q000378:metabolism; D053673:Glypicans / Q000276:immunology / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008027:Light; D008113:Liver Neoplasms / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D008279:Magnetic Resonance Imaging; D058185:Magnetite Nanoparticles; D046529:Microscopy, Electron, Transmission; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000138:chemical synthesis* / Q000378:metabolism; D010316:Particle Size; D011237:Predictive Value of Tests; D012542:Scattering, Radiation; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000911:Antibodies, Monoclonal; D003287:Contrast Media; D003911:Dextrans; C105937:GPC3 protein, human; D053673:Glypicans; D058185:Magnetite Nanoparticles; D015335:Molecular Probes; C097921:ferumoxtran-10
keywords: Hepatocellular carcinoma; Magnetic resonance imaging; One-pot method; Specific probe; Ultrasmall superparamagnetic iron oxide nanoparticles