Trastuzumab (Herceptin) Pharmacokinetic ELISA Kit
适用物种:Human,mouse,rat

样本类型:Serum,Plasma

检测范围:40ng/ml - 1.25ng/ml

最低检出限:1.25ng/ml

测定时间:2.5 hours

检测格式:Direct sandwich ELISA

检测原理:
The Trastuzumab ELISA kit is designed to measure free Trastuzumab with high specificity and sensitivity . This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Trastuzumab 96 well plate is provided. 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 and the concentration is calculated from the standard series.

检测方法:Peroxidase / OD450 

靶抗原:Trastuzumab (Herceptin)

应用:Quantification of Trastuzumab in biological matrices

包被板类型:Strip

样品体积:15ul

特异性:Trastuzumab

检测精度(批内差异,批间差异):<10%, <10%

防腐剂:None

Gene ID:2064

试剂盒组分:
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

储存温度和稳定性:Stable at -20°C for 1 year

背景描述:
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.

检测程序:
This assay employs the sandwich enzyme immunoassay technique. Anti- Trastuzumab is coated onto a 96 well microplate. Calibrator, quality control samples (if desired) and test samples are pipetted into the appropriate wells. Trastuzumab present in biological matrices is bound by the immobilized anti- Trastuzumab antibody. After washing away any unbound substances, enzyme linked antiTrastuzumab 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 color development is stopped and the intensity of the color is measured.

样本收集:
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.

样品制备:
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.

试剂制备:
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.

结果计算:
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.



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