Trastuzumab (Herceptin®)免疫原性ELISA试剂盒

抗原（Antigen）：Trastuzumab (Herceptin) Immunogenicity
反应性（Reactivity）：Human, mouse, rat
特异性（Specificity）：Anti-Trastuzumab antibodies
检测范围（Detection range）：125ng/ml - 31.25ng/ml
灵敏度（Sensitivity）：31.25ng/ml应用（Application）：Quantification of antibodies to Trastuzumab
检测类型（Method type）：Bridging 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）：3.5 hours
防腐剂（Preservative）：None
试剂盒组分（Components）：
Coated microtiter plate, 96 wells
QC samples - 4x50ul
10X wash buffer - 25ml
Assay buffer - 50ml
1000X secondary antibody - 17ul
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

曲妥珠单抗免疫原性ELISA试剂盒，Trastuzumab (Herceptin®) Immunogenicity ELISA Kit，可用于定性检测生物基质中抗曲妥珠单抗抗体的存在。曲妥珠单抗是一种人源化重组单克隆抗体，用于治疗过度表达人表皮生长因子受体2（HER2）的原发性乳腺癌。

背景信息（Background information）：
Trastuzumab (Herceptin® ) is a humanized recombinant monoclonal antibody used for the treatment of primary breast cancers overexpressing human epidermal growth factor 2 (HER2). HER2 protein is overexpressed in 25-30% of breast cancers.

检测原理（Summary of the assay）：
The Trastuzumab immunogenicity assay employs the bridging ELISA technique. A precoated 96 well capture antibody plate is provided. Quality control and test samples are pipetted into the appropriate wells. Anti-Trastuzumab present in biological matrices binds the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. 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 anti-Trastuzumab present in test samples. Four levels of QC samples give a qualitative reference signal which can be used to determine the level (High, Medium, Low, Negative) of anti-Trastuzumab antibody in the unknown samples.

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 QC samples and test samples 1/10 with assay buffer (for example add 30µL of prepared calibrator or sample to 270µL of assay buffer). Mix well. Do not store diluted samples. If test samples are out of range, then they may be further diluted.

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 ultra-pure water 1/10 before use (for example add 2mL concentrate to 18mL ultra-pure water). Mix well.
2. Secondary antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for examples add 12μl concentrate to 12ml of assay buffer). Mix well.
3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12μl concentrate to 12ml of assay buffer). Mix well.

检测程序（Assay procedure）：
This immunogenicity assay employs the bridging ELISA technique. Capture antibody is precoated onto a 96 well microplate. Quality control and test samples are pipetted into the appropriate wells. AntiTrastuzumab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, secondary antibody is added to the wells and after a final wash a detection reagent is added. 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 anti-Trastuzumab present in test samples. Three levels of QC samples give a qualitative reference signal which can be used to determine the level of anti-Trastuzumab antibody in the unknown samples. The color development is stopped and the intensity of the color is measured.

结果计算（Results calculation）：
1. Because anti-drug antibodies will vary in terms of affinity and concentration, this assay provides a qualitative readout. As such the user should use the comparable positive controls when comparing interassay results. The provided controls are tested for comparability between lots and can be traced.
2. The anti-drug antibody titers in the test samples will fall in the range of high, medium, low or negative. We recommend each lab develop their own statistical cutpoint using methodologies as described by G. Shankar, et al. (2008). (Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J. Pharmaceutical and Biomedical Analysis 48:1267–1281).
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]：勃谱生物的曲妥珠单抗免疫原性ELISA试剂盒(Trastuzumab 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.