Cygnus F550 CHO HCP ELISA Kit 宿主细胞蛋白免疫检测试剂盒，第三代

产品英文名称：CHO HCP ELISA kit, 3G
产品中文名称：CHO HCP酶联免疫分析试剂盒

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CHO HCP ELISA 试剂盒 产品介绍：
CHO HCP是第三代ELISA试剂盒，比之前试剂盒具有更广泛反应性。新型试剂盒在检测CHO HCPs 蛋白方面有很多改进。新试剂盒具有更高的灵敏度，在抗体物和其它CHO表达蛋白中检测HCP蛋白可以精确到到十亿分之一百。第三代ELISA试剂盒经过大量药物原料和中试样品检测评估，是工艺开发和大量释放试验检测的通用方法。抗体反应性使用传统2D 蛋白杂交技术（2D HPLC-ELISA）鉴定。2D HPLC-ELISA方法显示抗体对超过750种HCPs都具有反应性。用蛋白检测方法和ELISA正交试验表明这750种HCPs相当于98%总蛋白量。因此，这种抗体可以避免丢失任何重要的HCP蛋白。产品使用时一定要做好保护措施：0.5M硫磺酸有强腐蚀性，要避免接触眼睛、皮肤和衣物。检测前所有试剂都要平衡到室温。

§检测原理：CHO检测是一个双位点酶联免疫检测。样本中包含的CHO HCPs蛋白与标记有抗-CHO抗体的辣根过氧化物酶同时反应，反应发生在之前涂有一层吸附性抗-CHO蛋白抗体的微量滴定板中进行。免疫反构成为夹层式结构：固相抗休-HCP-酶标记抗体。反应结束后清洗微量滴定板去除未结合反应物。添加TMB作用底物反应。微量滴定板读数器上测定水解物浓度，水解物浓度与CHO HCPs蛋白浓度成比例。用提供的标准液绘制标准曲线，根据读数可以计算出CHO HCPs蛋白含量。

§检测过程：取出所有试剂平衡到室温→移取100µl anti-CHO:HRP（#F551）至每孔→分别移取50µl标准液，对照物和样品至待测孔→密封滴定板，摇床室温温育2小时，180rpm→清空小孔，用纸吸干残留液，350µl清洗缓冲液清洗4次→加100µl TMB 作用物（#F005）→室温静置30分钟，不需摇床→加100µl反应终止液（#F006）→分光光度计450/650nm读数

§结果分析：标准液（0, 1, 3, 12, 40, 100ng/mL）可用于制作标准曲线，标准液浓度相当于免疫反应HCP总浓度。根据测定的小孔中标准液吸光值绘制标准曲线。数据分析可以使用计算机选配曲线，例如点到点，三次样本函数，或四参数逻辑曲线。不要使用线性回归分析修改样品吸光值，这样会出现显著误差。也可以手工绘图，Y轴标注吸光值，X轴标注浓度，然后用光滑曲线连接各点。然后根据样品吸光值计算出样品中HCP浓度。（操作时每一浓度标准液都要滴定到两个小孔中，以便计算平均分光光度值，样品也需计算平均吸光值）。

CHO HCP ELISA组成成份：
● Anti-CHO:HRP（辣根过氧化物酶）   1x12mL   F551（Product#）
● Anti-CHO微量滴定板           12x8孔  F552
● CHO HCP标准液（0, 1, 3, 12, 40, 100ng/mL）  1mL/瓶 F553
● Stop Solution终止液（0.5M硫磺酸）   1x12mL F006
● TMB （3,3’,5,5’-四甲基联苯氨）           1x12mL F005
● Wash Concentrate清洗物（20倍浓缩）    1x50mL   F004

需自备物：
» 微量板分光光度计，双波长450 & 650nm
» 移液器-50L and 100L
» 可重复使用或多通道移液器-100L
» 微量板摇床（150 - 200 rpm）
» 样品稀释液（推荐使用Cat # I028）
» 1L清洗瓶
» 蒸馏水

CHO HCP ELISA kit, 3G英文简述：
This CHO HCP 3rd Generation ELISA kit uses a new and even more broadly reactive antibody than our earlier kits. The kit incorporates a number of other improvements in the detection of CHO HCPs. This new ELISA is more sensitive than the previous kits, able to detect HCPs in the range of 100 parts per billion for a variety of antibodies and other therapeutic proteins expressed in CHO. It has been extensively qualified with a large number of drug substances and in-process samples. This data indicates the kit is truly a generic method that can likely be validated as not only a process development tool, but in most cases as a lot release test, eliminating the need to develop a custom or process specific assay. The antibody has been evaluated for reactivity to individual HCPs using the conventional 2D Western Blot as well as a method we have shown to be superior to 2D WB, which we call 2D HPLC-ELISA. This 2D HPLC-ELISA method demonstrated the antibody is reactive to more than 750 individual HCPs from conditioned media and cell lysates in both a CHO-S strain as well as a K1 strain. These 750+ HCPs represent more than 98% of the total mass of protein as indicated by protein detection methods orthogonal to ELISA. As such, this antibody may overcome concerns about missing any significant individual HCPs. This new ELISA is also more robust than the earlier kits showing less sample matrix interference and improved dilutional linearity. If you have found the F015 and CM015 kits lacking in any way meeting your HCP detection needs, or if you are considering the need for a process-specific assay, we recommend you evaluate this 3rd Generation CHO HCP ELISA kit.

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