產(chǎn)品英文名稱：CHO HCP ELISA kit, 3G
產(chǎn)品中文名稱：CHO HCP酶聯(lián)免疫分析試劑盒

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CHO HCP ELISA 試劑盒 產(chǎn)品介紹：
CHO HCP是第三代ELISA試劑盒，比之前試劑盒具有更廣泛反應(yīng)性。新型試劑盒在檢測CHO HCPs 蛋白方面有很多改進(jìn)。新試劑盒具有更高的靈敏度，在抗體物和其它CHO表達(dá)蛋白中檢測HCP蛋白可以精確到到十億分之一百。第三代ELISA試劑盒經(jīng)過大量藥物原料和中試樣品檢測評估，是工藝開發(fā)和大量釋放試驗(yàn)檢測的通用方法?？贵w反應(yīng)性使用傳統(tǒng)2D 蛋白雜交技術(shù)（2D HPLC-ELISA）鑒定。2D HPLC-ELISA方法顯示抗體對超過750種HCPs都具有反應(yīng)性。用蛋白檢測方法和ELISA正交試驗(yàn)表明這750種HCPs相當(dāng)于98%總蛋白量。因此，這種抗體可以避免丟失任何重要的HCP蛋白。產(chǎn)品使用時(shí)一定要做好保護(hù)措施：0.5M硫磺酸有強(qiáng)腐蝕性，要避免接觸眼睛、皮膚和衣物。檢測前所有試劑都要平衡到室溫。

§檢測原理：CHO檢測是一個(gè)雙位點(diǎn)酶聯(lián)免疫檢測。樣本中包含的CHO HCPs蛋白與標(biāo)記有抗-CHO抗體的辣根過氧化物酶同時(shí)反應(yīng)，反應(yīng)發(fā)生在之前涂有一層吸附性抗-CHO蛋白抗體的微量滴定板中進(jìn)行。免疫反構(gòu)成為夾層式結(jié)構(gòu)：固相抗休-HCP-酶標(biāo)記抗體。反應(yīng)結(jié)束后清洗微量滴定板去除未結(jié)合反應(yīng)物。添加TMB作用底物反應(yīng)。微量滴定板讀數(shù)器上測定水解物濃度，水解物濃度與CHO HCPs蛋白濃度成比例。用提供的標(biāo)準(zhǔn)液繪制標(biāo)準(zhǔn)曲線，根據(jù)讀數(shù)可以計(jì)算出CHO HCPs蛋白含量。

§檢測過程：取出所有試劑平衡到室溫→移取100µl anti-CHO:HRP（#F551）至每孔→分別移取50µl標(biāo)準(zhǔn)液，對照物和樣品至待測孔→密封滴定板，搖床室溫溫育2小時(shí)，180rpm→清空小孔，用紙吸干殘留液，350µl清洗緩沖液清洗4次→加100µl TMB 作用物（#F005）→室溫靜置30分鐘，不需搖床→加100µl反應(yīng)終止液（#F006）→分光光度計(jì)450/650nm讀數(shù)

§結(jié)果分析：標(biāo)準(zhǔn)液（0, 1, 3, 12, 40, 100ng/mL）可用于制作標(biāo)準(zhǔn)曲線，標(biāo)準(zhǔn)液濃度相當(dāng)于免疫反應(yīng)HCP總濃度。根據(jù)測定的小孔中標(biāo)準(zhǔn)液吸光值繪制標(biāo)準(zhǔn)曲線。數(shù)據(jù)分析可以使用計(jì)算機(jī)選配曲線，例如點(diǎn)到點(diǎn)，三次樣本函數(shù)，或四參數(shù)邏輯曲線。不要使用線性回歸分析修改樣品吸光值，這樣會出現(xiàn)顯著誤差。也可以手工繪圖，Y軸標(biāo)注吸光值，X軸標(biāo)注濃度，然后用光滑曲線連接各點(diǎn)。然后根據(jù)樣品吸光值計(jì)算出樣品中HCP濃度。（操作時(shí)每一濃度標(biāo)準(zhǔn)液都要滴定到兩個(gè)小孔中，以便計(jì)算平均分光光度值，樣品也需計(jì)算平均吸光值）。

CHO HCP ELISA組成成份：
● Anti-CHO:HRP（辣根過氧化物酶）   1x12mL   F551（Product#）
● Anti-CHO微量滴定板           12x8孔  F552
● CHO HCP標(biāo)準(zhǔn)液（0, 1, 3, 12, 40, 100ng/mL）  1mL/瓶 F553
● Stop Solution終止液（0.5M硫磺酸）   1x12mL F006
● TMB （3,3’,5,5’-四甲基聯(lián)苯氨）           1x12mL F005
● Wash Concentrate清洗物（20倍濃縮）    1x50mL   F004

需自備物：
? 微量板分光光度計(jì)，雙波長450 & 650nm
? 移液器-50L and 100L
? 可重復(fù)使用或多通道移液器-100L
? 微量板搖床（150 - 200 rpm）
? 樣品稀釋液（推薦使用Cat # I028）
? 1L清洗瓶
? 蒸餾水

CHO HCP ELISA kit, 3G英文簡述：
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.

重慶市華雅干細(xì)胞技術(shù)有限公司