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西妥昔单抗(Cetuximab)是针对EGF受体的IgG1单克隆抗体,两者特异性结合后,通过对与EGF受体结合的酪氨酸激酶(TK)的抑制作用,阻断细胞内信号转导途径,从而抑制癌细胞的增殖,诱导癌细胞的凋亡,减少基质金属蛋白酶和血管内皮生长因子的产生。在美国,西妥昔单抗用于治疗头颈癌和结直肠癌。在欧盟,西妥昔单抗适用于治疗表达表皮生长因子受体(EGFR) 的 RAS 野生型转移性结直肠癌和头颈部鳞状细胞癌。 本试剂盒采用间接法酶联免疫吸附试验(ELISA)。往预先包被有人EGFR抗原的微孔中,依次加入样本、标准品、HRP标记的二抗,中间经过温育和洗涤,用底物TMB显色,TMB在过氧化物酶(HRP)的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样本中的西妥昔单抗(Cetuximab)呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
This kit employs the indirect enzyme-linked immunosorbent assay (ELISA) principle. Microplate wells are pre-coated with Human EGFR antigen. The sample or standard is first added to the wells, followed by an HRP (horseradish peroxidase)-labeled secondary antibody. The procedure involves incubation and wash steps between each addition. TMB substrate is then added for color development. Under the catalysis of the peroxidase (HRP), TMB is converted into a blue product, which subsequently turns into a final stable yellow color upon the addition of an acid stop solution. The intensity of the yellow color is directly proportional to the concentration of Cetuximab in the sample. The optical density (OD) is measured at a wavelength of 450 nm using a microplate reader, and the concentration of the target analyte in the sample is calculated based on the standard curve.