title: Direct chemiluminescent imaging detection of human serum proteins in two-dimensional polyacrylamide gel electrophoresis.
abstract: A novel chemiluminescence (CL)-based imaging method capable of directly detecting proteins in polyacrylamide gels after electrophoresis is proposed. Human serum proteins are presently detected by a direct CL imaging method after native 2-D PAGE. As a consequence, some proteins, including haptoglobin (Hp), Hp precursor, hemopexin (Hpx) precursor, Ig alpha-1 chain C region, and Complement C3 precursor can be detected and identified by MS and MS/MS techniques. These proteins are all acute phase proteins, which have been defined as biomarkers for certain diseases. Moreover, serum proteins from healthy people and cirrhotic patients were analyzed. A decrease in Hp spots for cirrhotic patients could be confirmed. The CL imaging conditions were optimized, including the concentrations of H(2)O(2) and luminol. The process of CL detection of proteins is simple, and there is no need for specialized equipment. In comparison with the traditional CBB-R250 staining method, the detection sensitivity was improved and the detection period decreased about 70 times. Hence, this technique possesses potentials as a rapid, convenient, and inexpensive analytical technique for protein detection and for the diagnosis of diseases.
authors.affiliation: College of Chemistry, Beijing Normal University, Beijing, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Tao
authors.forename: Zhenzhen
authors.initials: Z
authors.lastname: Wang
authors.forename: Heping
authors.initials: H
authors.lastname: Zhao
authors.forename: Willy R G
authors.initials: WR
authors.lastname: Baeyens
authors.forename: Joris R
authors.initials: JR
authors.lastname: Delanghe
authors.forename: Lingyun
authors.initials: L
authors.lastname: Huang
authors.forename: Jin
authors.initials: J
authors.lastname: Ouyang
authors.forename: Dacheng
authors.initials: D
authors.lastname: He
authors.forename: Xianhua
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D015415:Biomarkers / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D013194:Staining and Labeling
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D010455:Peptides