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.
OBJECTIVES: To validate a specific enzyme-linked immunosorbent assay for the rapid-acting humaninsulin 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, humanproinsulin, or humanC-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: Christian Rask-Madsen; Erica Buonomo; Qian Li; Kyoungmin Park; Allen C Clermont; Oluwatobi Yerokun; Mark Rekhter; George L King Journal: Arterioscler Thromb Vasc Biol Date: 2012-03-15 Impact factor: 8.311
Authors: D R McCance; P Damm; E R Mathiesen; M Hod; R Kaaja; F Dunne; L E Jensen; H Mersebach Journal: Diabetologia Date: 2008-08-23 Impact factor: 10.122
Authors: Tina Parkner; Torben Laursen; Jian-Wen Chen; Marianne K Møller; Henrik F Thomsen; Christina Jørgensen; Jørgen S Smedegaard; Torsten Lauritzen; Jens S Christiansen Journal: J Diabetes Sci Technol Date: 2007-09
Authors: Lindy Kahanovitz; Erkin Seker; Robert S Marks; Martin L Yarmush; Tania Konry; Steven J Russell Journal: J Diabetes Sci Technol Date: 2016-05-03