A new insulin immunoassay specific for the rapid-acting insulin analog, insulin aspart, suitable for bioavailability, bioequivalence, and pharmacokinetic studies

Clin Biochem. 2000 Nov;33(8):627-33. doi: 10.1016/s0009-9120(00)00183-1.


Objectives: 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.

Design and 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.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Biological Availability
  • Cross Reactions
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Insulin / analogs & derivatives*
  • Insulin / analysis*
  • Insulin / immunology
  • Insulin Aspart
  • Mice
  • Pharmacokinetics
  • Radioimmunoassay
  • Reproducibility of Results
  • Swine
  • Therapeutic Equivalency


  • Antibodies, Monoclonal
  • Insulin
  • Insulin Aspart