title: Specificity and sensitivity of commercially available assays for glucagon and oxyntomodulin measurement in humans.
abstract: OBJECTIVE
  To determine the specificity and sensitivity of assays carried out using commercially available kits for glucagon and/or oxyntomodulin measurements.
  
  
  METHODS
  Ten different assay kits used for the measurement of either glucagon or oxyntomodulin concentrations were obtained. Solutions of synthetic glucagon (proglucagon (PG) residues 3361), oxyntomodulin (PG residues 3369) and glicentin (PG residues 169) were prepared and peptide concentrations were verified by quantitative amino acid analysis and a processing-independent in-house RIA. Peptides were added to the matrix (assay buffer) supplied with the kits (concentration range: 1.25-300 pmol/l) and to human plasma and recoveries were determined. Assays yielding meaningful results were analysed for precision and sensitivity by repeated analysis and ability to discriminate low concentrations.
  
  
  CONCLUSIONS
  Three assays were specific for glucagon (carried out using the Millipore (Billerica, MA, USA), Bio-Rad (Sundbyberg, Sweden), and ALPCO (Salem, NH, USA) and Yanaihara Institute (Shizuoka, Japan) kits), but none was specific for oxyntomodulin. The assay carried out using the Phoenix (Burlingame, CA, USA) glucagon kit measured the concentrations of all three peptides (total glucagon) equally. Sensitivity and precision were generally poor; the assay carried out using the Millipore RIA kit performed best with a sensitivity around 10 pmol/l. Assays carried out using the BlueGene (Shanghai, China), USCN LIFE (Wuhan, China) (oxyntomodulin and glucagon), MyBioSource (San Diego, CA, USA) and Phoenix oxyntomodulin kits yielded inconsistent results.
authors.affiliation: NNF Center for Basic Metabolic Research, Department of Biomedical Sciences, Faculty of Health Sciences.
authors.forename: Monika J
authors.initials: MJ
authors.lastname: Bak
authors.forename: Nicolai Wewer
authors.initials: NW
authors.lastname: Albrechtsen
authors.forename: Jens
authors.initials: J
authors.lastname: Pedersen
authors.forename: Bolette
authors.initials: B
authors.lastname: Hartmann
authors.forename: Mikkel
authors.initials: M
authors.lastname: Christensen
authors.forename: Tina
authors.initials: T
authors.lastname: Vilsbøll
authors.forename: Filip K
authors.initials: FK
authors.lastname: Knop
authors.forename: Carolyn F
authors.initials: CF
authors.lastname: Deacon
authors.forename: Lars O
authors.initials: LO
authors.lastname: Dragsted
authors.forename: Jens J
authors.initials: JJ
authors.lastname: Holst
mesh_terms: D003924:Diabetes Mellitus, Type 2 / Q000175:diagnosis; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D005934:Glucagon / Q000032:analysis* / Q000097:blood; D006801:Humans; D053772:Oxyntomodulin / Q000032:analysis* / Q000097:blood; D011863:Radioimmunoassay / Q000592:standards; D012680:Sensitivity and Specificity
chemical_list: D053772:Oxyntomodulin; D005934:Glucagon