A new insulin immunoassay specific for the rapid-acting insulin analog, insulin aspart, suitable for bioavailability, bioequivalence, and pharmacokinetic studies.
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.
Andersen
L
L
Novo Nordisk A/S, Hagedornsveg 1 (HAB3.66), DK-2820 Copenhagen, Denmark. lean@novonordisk.com
Jørgensen
P N
PN
Jensen
L B
LB
Walsh
D
D
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
D000911:Antibodies, Monoclonal; D007328:Insulin; D061267:Insulin Aspart