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三分餓的吧至尊木蟲 (著名寫手)
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[求助]
求幫忙翻譯下這篇英文文獻(xiàn)摘要~,生化與分子方面的
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| A direct competitive enzyme-linked immunosorbent assay (dcELISA) based on a monoclonal antibody was developed for detecting a new beta-adrenergic agonist phenylethanolamine A (PEAA). A PEAA derivative, PEAA-NH2, was synthesized by hydrogenation using RANEY (R) nickel as a catalyst, before it was coupled to carrier proteins to generate PEAA-BSA, PEAA-OVA and PEAA-HRP via diazotization and sodium periodate methods, repectively. Three Balb/C mice were immunized with PEAA-BSA as the immunogen, and three clones stably producing antibodies against PEAA were acquired. Subsequently, a dcELISA method was established using the monoclonal antibody (clone 2H8) and PEAA-HRP. With the PEAA concentration range of 0.025-2.025 ng mL(-1), the IC50 value was 0.204 ng mL(-1). The monoclonal antibody was highly specific for PEAA and its derivative (PEAA-NH2), with minor cross-reactivity (CR) with ractopamine (CR = 8.3%) and negligible cross-reactivity with 15 other beta-agonists compounds (CR < 0.1%). For sample testing, the LOD (limit of detection) values of blank samples (pork, swine liver, swine urine and feed) were shown to be 0.431, 0.455, 0.225 and 8.693 ng mL(-1), respectively; the corresponding LOQ (limit of quantification) values were 0.578, 0.647, 0,360 and 14.477 ng mL(-1), respectively. The recovery rates ranged from 82% to 120%. The coefficients of variation were below 13.33% and 10.53% for inter-assay and intra-assay, respectively. Furthermore, the dcELISA was validated by a liquid chromatography tandem mass spectrometry (LC-MS/MS) method, and the result showed a high correlation coefficient between the two methods. Overall, the results suggested this dcELISA method with high sensitivity and specificity could be used to detect PEAA residues in real samples reliably. |
至尊木蟲 (著名寫手)
| 建立了一種基于單克隆抗體的直接競爭性酶聯(lián)免疫吸附試驗 (dcELISA)用于檢測新的beta-腎上腺素能激活劑苯乙醇胺A (PEAA).在分別通過重氮反應(yīng)和過碘酸鈉法將其偶聯(lián)到載體蛋白產(chǎn)生PEAA-BSA,PEAA-OVA和PEAA-HRP前,以 RANEY (R)鎳為催化劑合成了一種PEAA的衍生物 PEAA-NH2。以PEAA-BSA為免疫原免疫3只Balb/C小鼠,獲得了穩(wěn)定產(chǎn)生對PEAA抗體的3個克隆。隨后,用單克隆抗體(2H8)和PEAA-HRP建立了dcELISA方法。PEAA濃度在0.025-2.025ng/mL范圍內(nèi),IC50值為0.204ng/mL。該單克隆抗體對PEAA及其衍生物PEAA-NH2高度特異,對萊克多巴胺有少量交叉反應(yīng)(CR)(CR = 8.3%),而對15種其它的beta-激活劑化合物的交叉反應(yīng)可忽略(CR < 0.1%)。對樣品測試,空白樣品(豬肉、豬肝、豬尿和飼料)的LOD (檢測限)值分別為0.431,0.455,0.225和8.693ng/mL;對應(yīng)的LOQ(定量限)值分別0.578,0.647,0,360和14.477ng/mL。 回收率范圍從82%到120%。批間和批內(nèi)變異系數(shù)分別為13.33%和10.53%以下。此外dcELISA還用了液相色譜偶聯(lián)質(zhì)譜(LC-MS/MS)法評價,結(jié)果顯示二法有高相關(guān)系數(shù)?傊Y(jié)果提示此 dcELISA以高靈敏度和特異性可用于可靠檢測真實樣品中PEAA的殘留。 |
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