Comparison of a luminescent oxygen channeling immunoassay and an ELISA for detecting insulin aspart in human serum.
The study was a comparison between a Luminescent Oxygen Channeling Immunoassay (LOCI) and an enzyme-linked immunosorbent assay (ELISA) for quantification of Insulin Aspart (IAsp) in human serum. The advantage of LOCI compared to ELISA is reduced workload and higher throughput. The ELISA assay was performed as published (Andersen et al., 2000 [5]). The LOCI followed a 2-step reaction. First, the sample was incubated for 1h with a mixture of biotinylated antibody specific for IAsp and beads coated with insulin-detecting antibody. This step was followed by a 30-min incubation with beads covalently coated with streptavidin. When the beads were brought in proximity through binding of IAsp, light was generated from a chemiluminescent reaction in the beads. This light was measured and quantified. Spiked samples with different concentrations of IAsp were prepared in human serum to compare ELISA and LOCI. Human serum samples (n=510) from a pilot study with healthy subjects receiving IAsp were also analysed and compared in the two assays. Higher precision, improved accuracy and a wider analytical range were found using LOCI compared to ELISA. However, sample haemolysis interfered more when using LOCI than ELISA. The IAsp concentrations determined in the human serum samples from the pilot study gave a good correlation between the two assays. In conclusion, LOCI can determine IAsp in human serum just as well as ELISA. Using LOCI reduces the workload, which is particularly useful when handling large sample sizes.
Petersen
Signe Beck
SB
Novo Nordisk A/S, Global Development and Diabetes Research Unit, Novo Nordisk Park, 2760 Måløv, Denmark.
Lovmand
Julie Mangor
JM
Honoré
Lone
L
Jeppesen
Claus Bekker
CB
Pridal
Lone
L
Skyggebjerg
Ole
O
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007004:Hypoglycemic Agents / Q000097:blood*; D007118:Immunoassay / Q000379:methods; D007328:Insulin / Q000031:analogs & derivatives* / Q000097:blood; D061267:Insulin Aspart; D008163:Luminescent Measurements / Q000379:methods*; D010100:Oxygen / Q000737:chemistry; D015203:Reproducibility of Results; D013552:Swine
D007004:Hypoglycemic Agents; D007328:Insulin; D061267:Insulin Aspart; D010100:Oxygen