title: Detection of cystatin C biomarker for clinical measurement of renal disease by developed ELISA diagnostic kits.
abstract: BACKGROUND
  Human cystatin C (HCC) is a potential biomarker for tubular damage and impaired renal function. It is difficult to obtain efficient paired monoclonal antibodies against HCC with low molecular to meet the requirements for clinical application The present study was to establish a stable and repeatable measurement for HCC with self-made monoclonal antibodies (McAbs) and Variable domain of heavy chain of heavy-chain antibody (VHHs) increase the sensitivity.
  
  
  METHODS
  With hybridoma technology and phage display technology: R-HCC as a screening antigen and N-HCC as the detector for antigens to obtain the specific antibody and established an enzyme-linked immunosorbent assay for human cystatin C using self-made McAbs and VHHs.
  
  
  RESULTS
  We have successfully obtained three McAbs; 5 F2, 4E4, 1E11 and four VHHs; 3-2, 3-24, 3-33 and 4-5 which were specific for HCC. The measurement of HCC was established with the self-made monoclonal antibodies and VHHs with a high sensitivity the lower limit of detection at 0.5 ng/ml and the detection range at 0.5 ~ 31.3 ng/ml.
  
  
  CONCLUSIONS
  Our data provides a new approach for paired antibody screening and testing of the small molecular biomarker with a single dominant epitope, with the important biological and clinical significance.
authors.forename: Renren
authors.initials: R
authors.lastname: Jiang
authors.forename: Chao
authors.initials: C
authors.lastname: Xu
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Key Laboratory for Food Safety Research, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. tianhao74@hotmail.com.
authors.forename: Tianhao
authors.initials: T
authors.lastname: Wang
authors.forename: Gang
authors.initials: G
authors.lastname: Yao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D055316:Cystatin C / Q000737:chemistry / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006639:Histidine; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D007674:Kidney Diseases / Q000175:diagnosis*; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009842:Oligopeptides; D011933:Reagent Kits, Diagnostic; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D015415:Biomarkers; D055316:Cystatin C; C471213:His-His-His-His-His-His; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D009842:Oligopeptides; D011933:Reagent Kits, Diagnostic; D011993:Recombinant Fusion Proteins; D006639:Histidine