AffinityImmuno · ELISA Kit
Trastuzumab(Herceptin®) Pharmacokinetic ELISA
Cat.No
EL-1611-201
1X96 wells
- Reactivity
- Human, mouse, rat
- Detection range
- 40ng/ml - 1.25ng/ml
- Sample type
- Serum Plasma
- Assay time
- 2.5 hours
- Antigen
- Trastuzumab (Herceptin)
- Detection minimum
- 1.25ng/ml
Product Overview
This Trastuzumab (Herceptin) ELISA Pharmacokinetic kit is designed to measure trastuzumab with high specificity and enhanced sensitivity.
INTRODUCTION
Trastuzumab (trade name Herceptin®) is indicated for the treatment of HER2-positive breast cancer, and adjuvant therapies for metastatic gastric cancer and gastroesophageal cancer. Serum concentration of trastuzumab may predict some clinical outcome during therapy. It is also possible that the surveillance of circulating trastuzumab concentration during therapy represents a direct factor for immunogenicity and some other side effects. Identification of biomarkers and risk factors for adverse drug reactions that might be related to serum concentrations, and maintaining the effective minimum concentration of trastuzumab in order to potentially avoid some side effects, might be beneficial using a reliable method.
The Trastuzumab ELISA kit is designed to measure trastuzumab with high specificity and enhanced sensitivity. The assay design utilizes a pair of antibodies that allows the detection of the whole trastuzumab molecule in biological matrices.
PRINCIPLE OF THE ASSAY
This assay employs the sandwich enzyme immunoassay technique. Anti-Trastuzumab is coated onto a 96 well microplate. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Trastuzumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, enzyme linked detection antibody is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Trastuzumab present in test samples.
The standard curve was generated with the supplied calibrator using the recommended
conditions. Each sample was run with 6 replicates. Intra-assay coefficient of variance is <10%.
Target background
Trastuzumab (Herceptin®) is indicated for the treatment of HER2-positive breast cancer, and adjuvant therapies for metastatic gastric cancer and gastroesophageal cancer. Serum concentration of Trastuzumab may predict some clinical outcome during therapy. It is also possible that the surveillance of circulating Trastuzumab concentration during therapy represents a direct factor for immunogenicity and some other side effects. Identification of biomarkers and risk factors for adverse drug reactions that might be related to serum concentrations, and maintaining the effective minimum concentration of Trastuzumab in order to potentially avoid some side effects, might be beneficial using a reliable method.
Specifications
- Gene ID
- 2064
- Reactivity
- Human, mouse, rat
- Detection range
- 40ng/ml - 1.25ng/ml
- Sample type
- Serum Plasma
- Sample volume
- 15ul
- Assay time
- 2.5 hours
- Assay precision
- <10%, <10%
- Storage
- -20C, 1 year
- Size
- 1X96 wells
- Antigen
- Trastuzumab (Herceptin)
- Detection minimum
- 1.25ng/ml
- Method
- Direct sandwich ELISA
- Detection method
- Peroxidase / OD450
- Application
- Quantification of Trastuzumab in biological matrices
- Plate format
- Strip
- Specificity
- Trastuzumab
- Preservative
- None
Kit Components
- Coated microtiter plate, 96 wells
- Calibrator diluent. - 1.8ml
- Calibrator 12ul
- 10X wash buffer - 25ml
- Assay buffer - 50ml
- 1000X detection reagent - 17ul
- TMB - 12ml
- TMB stop solution - 12ml
- Plate sealers - 3
Assay Information
Reagent preparation
Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/10 before use (for example add 10mL concentrate to 90mL ultra-pure water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 10μl concentrate to 10ml of assay buffer). Mix welll. 3. Preparation of Calibrators: Prepare calibrators with concentrations ranging from 1000 ng/mL to 62.5 ng/mL. The following is an example calibrator curve.
Results calculation
1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. Alternatively a log-log curve fit may be used. 2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample. 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with calibrator diluent and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
Sample collection
This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20°C for up to 1 year.
Sample preparation
Dilute calibrators and test samples 1/50 with assay buffer (for example add 5µL of prepared calibrator or sample to 245µL of assay buffer). Mix well. Do not store diluted samples.