title: A new insulin immunoassay specific for the rapid-acting insulin analog, insulin aspart, suitable for bioavailability, bioequivalence, and pharmacokinetic studies.
abstract: OBJECTIVE
  To validate a specific enzyme-linked immunosorbent assay for the rapid-acting human insulin analogue, insulin aspart, in human serum, human plasma, and porcine plasma.
  
  
  METHODS
  For the enzyme-linked immunosorbent assay, two murine monoclonal antibodies were developed that bind to two different epitopes on the insulin aspart molecule. Key parameters for validation were imprecision, accuracy, matrix effects, dilution-linearity, and cross-reactivity.
  
  
  RESULTS
  No cross-reactivity was found with human and porcine insulin, human proinsulin, or human C-peptide. The assay is sensitive (limit of quantification = 11.5 pmol/L), accurate (95-107% recovery with human serum, human plasma, and porcine plasma in the range 16-800 pmol/L), and has a 14.7% total imprecision within the entire analytical range. Dilution of samples gave linear results with human serum as the diluent.
  
  
  CONCLUSIONS
  The insulin aspart-specific enzyme-linked immunosorbent assay described in this study is well suited to study the bioavailability, bioequivalence, and pharmacokinetics of this insulin analogue.
authors.affiliation: Novo Nordisk A/S, Hagedornsveg 1 (HAB3.66), DK-2820 Copenhagen, Denmark. lean@novonordisk.com
authors.forename: L
authors.initials: L
authors.lastname: Andersen
authors.forename: P N
authors.initials: PN
authors.lastname: Jørgensen
authors.forename: L B
authors.initials: LB
authors.lastname: Jensen
authors.forename: D
authors.initials: D
authors.lastname: Walsh
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001682:Biological Availability; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives* / Q000032:analysis* / Q000276:immunology; D061267:Insulin Aspart; D051379:Mice; D010599:Pharmacokinetics; D011863:Radioimmunoassay; D015203:Reproducibility of Results; D013552:Swine; D013810:Therapeutic Equivalency
chemical_list: D000911:Antibodies, Monoclonal; D007328:Insulin; D061267:Insulin Aspart