Human g csf duoset elisa kit

Blood samples for PK analysis were collected during the in‐patient phase, predose and up to 96 hours postdose, and during the ambulatory visits in each period.
Pegfilgrastim concentrations in serum were determined using a standard quantitative enzyme‐linked immunosorbent assay (ELISA) technique. The assay employed components from the R&D Systems’ (Biotechne AG, Switzerland) Human G‐CSF DuoSet ELISA kit. Microplates are coated with mouse anti‐human G‐CSF capture antibody which binds the G‐CSF in the sample. After the analyte is bound it is detected using a biotinylated goat anti‐human G‐CSF detection antibody. The capture antibody is then bound by streptavidin‐horseradish‐peroxidase (HRP), which in turn enzymatically catalyses tetramethylbenzidine (TMB) conversion.
The determination was carried out over an expected calibration range of 0.20‐8.00 ng/mL (samples above the calibration range could be diluted up to 400‐fold). The lower limit of quantification of the assay was 0.2 ng/mL For low, medium, and high quality control, the intraday variability was between 3.9% and 6.5% while interday variability was between 3.1% and 4.3%. The method was validated in accordance with the European Medicines Agency (EMA) Guideline on Bioanalytical Method Validation9 and the FDA Draft Guidance for Industry on Bioanalytical Method Validation 10.