Cetuximab (Erbitux®)药代动力学ELISA试剂盒

抗原（Antigen）：Cetuximab (Erbitux)
反应性（Reactivity）：Human, mouse, rat
特异性（Specificity）：Cetuximab
检测范围（Detection range）：50ng/ml - 1.56ng/ml
灵敏度（Sensitivity）：1.5ng/ml应用（Application）：Quantification of Cetuximab in biological matrices
检测类型（Method type）：Direct sandwich ELISA
检测方法（Detection Method）：Peroxidase / OD450 
检测精度（Assay precision）：Intra-assay coefficient of variation (CV) <10%. Inter-assay CV was <10%
酶标板类型（Plate）：Strip
样本类型（Sample type）：Serum and Plasma
样本体积（Sample volume）：15ul
检测时间（Assay time）：2.5 hours
防腐剂（Preservative）：None
试剂盒组分（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
运输条件（Shipping）：Dry ice干冰
储存（Storage）：Stable at -20℃ for 1 year

西妥昔单抗药代动力学ELISA试剂盒，Cetuximab (Erbitux®) Pharmacokinetic ELISA Kit，包含一种嵌合型 IgG1 单克隆抗体，该抗体可与表皮生长因子受体（EGFR）的胞外域相结合。适用于血浆和血浆样本。

背景信息（Background information）：
Cetuximab (Erbitux®) is a chimeric IgG1 monoclonal antibody that binds the extra-cellular domain of the epidermal growth factor receptor (EGFR). It is a 152-kDa molecule composed of four polypeptide chains: two identical heavy chains and two identical light chains, consisting of 449 and 214 amino acids, respectively, bound by covalent and non-covalent bonds. The bond with EGFR is characterized by a higher affinity than either endogenous ligand, as epidermal growth factor (EGF), or transforming growth factor alpha. This binding inhibits activation of the receptor tyrosine kinase and the associated downstream signaling that includes the mitogenactivated protein kinase, phosphoinositide 3-kinase/ Akt and the Janus kinases/ signal transducers and activator of transcription (Stat) pathways. Furthermore Cetuximab induces antibody-mediated receptor dimerization, internalization and degradation leading to receptor down-regulation. In addition, it exhibits antibody-dependent cellular cytotoxicity that could contribute to its antitumor effect.

检测原理（Summary of the assay）：
The Cetuximab ELISA kit is designed to measure free Cetuximab with high specificity and sensitivity . This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Cetuximab 96 well plate is provided. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Cetuximab 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 Cetuximab present in test samples and the concentration is calculated from the standard series.

样本采集（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.

样本制备（Sample preparation）：
Dilute calibrators and test samples 1/100 with assay buffer (for example add 5µL of prepared calibrator or sample to 495µL of assay buffer). Mix well. Do not store diluted samples.

试剂准备（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 deionized water 1/10 before use (for example add 20mL concentrate to 180mL deionized water). Mix well.
2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11μl concentrate to 11ml of assay buffer). Mix well. The following is an example calibrator curve.

检测程序（Assay procedure）：
This assay employs the sandwich enzyme immunoassay technique. Anti- Cetuximab is coated onto a 96 well microplate. Calibrator, quality control samples (if desired) and test samples are pipetted into the appropriate wells. Cetuximab present in biological matrices is bound by the immobilized anti- Cetuximab antibody. After washing away any unbound substances, enzyme linked anti- Cetuximab antibody is added to the wells. This antibody is developed and purified specifically against truncated Erbitux® (domain residing in Fc portion of the Erbitux® molecule). 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 Cetuximab present in test samples. The color development is stopped and the intensity of the color is measured.

结果计算（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 assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.

[注意事项]：勃谱生物的西妥昔单抗PK ELISA试剂盒(Cetuximab PK ELISA试剂盒)产品仅用于研究用途。不可用于诊断、医疗或其他用途。
[NOTICE]: Our products are for RESEARCH USE ONLY. Not for diagnostic, medical or other use.