title: The application of Au nanoclusters in the fluorescence imaging of human serum proteins after native PAGE: enhancing detection by low-temperature plasma treatment.
abstract: Proteins in human serum are increasingly being studied for their roles in a wide variety of biochemical interactions. To improve the sensitivity of the detection of human serum proteins after native polyacrylamide gel electrophoresis (PAGE), we have developed a fluorescence imaging detection technique for the detection. BSA (bovine serum albumin)-stabilized Au nanoclusters (NCs) were applied as fluorescent probes for imaging, and low-temperature plasma (LTP) treatment of the Au NCs was introduced to enhance the fluorescence imaging. Here, a series of optimization experiments (e.g. those to optimize for pH) were conducted for protein detection after 1-DE and 2-DE, and several types of discharge gases (He, O(2), and N(2)) were selected for the LTP treatment. The possible mechanism of interaction between the proteins and the Au NCs was demonstrated by an isothermal titration calorimetry experiment. Using the present method, a sensitivity of 7-14 times higher than that of traditional staining detection methods was observed in the oxygen LTP-treated Au NCs fluorescence images, and some relatively low abundance proteins (identified by the MS/MS technique) were easily detected. In addition, this fluorescence imaging method was applied to distinguish between the serum samples of patients with liver diseases and those of healthy people. Thus, this fluorescence imaging method is suitable for the highly sensitive detection of various serum proteins, and it shows potential capabilities for clinical diagnosis.
authors.affiliation: College of Chemistry, Beijing Normal University, Beijing 100875, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: College of Chemistry, Beijing Normal University, Beijing 100875, PR China.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Sajid
authors.affiliation: College of Chemistry, Beijing Normal University, Beijing 100875, PR China.
authors.forename: Na
authors.initials: N
authors.lastname: Na
authors.affiliation: Key Laboratory for Cell Proliferation and Regulation Biology, Ministry of Education, Beijing Normal University, Beijing 100875, PR China.
authors.forename: Lingyun
authors.initials: L
authors.lastname: Huang
authors.affiliation: Key Laboratory for Cell Proliferation and Regulation Biology, Ministry of Education, Beijing Normal University, Beijing 100875, PR China.
authors.forename: Dacheng
authors.initials: D
authors.lastname: He
authors.affiliation: College of Chemistry, Beijing Normal University, Beijing 100875, PR China. Electronic address: jinoyang@bnu.edu.cn.
authors.forename: Jin
authors.initials: J
authors.lastname: Ouyang
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods* / Q000706:statistics & numerical data; D001798:Blood Proteins / Q000032:analysis*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D006046:Gold; D006801:Humans; D008107:Liver Diseases / Q000097:blood / Q000175:diagnosis; D053768:Metal Nanoparticles; D012710:Serum Albumin, Bovine; D053719:Tandem Mass Spectrometry; D013696:Temperature
chemical_list: D001798:Blood Proteins; D005456:Fluorescent Dyes; D012710:Serum Albumin, Bovine; D006046:Gold