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三分饿的吧至尊木虫 (著名写手)
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[求助]
求帮忙翻译下这篇英文文献摘要~，生化与分子方面的
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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. |
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baoshanqiu
至尊木虫 (著名写手)
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- 专业: 胃肠道免疫相关疾病
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三分饿的吧: 金币+140, 翻译EPI+1, ★★★★★最佳答案 2016-01-06 09:20:54
三分饿的吧: 金币+140, 翻译EPI+1, ★★★★★最佳答案 2016-01-06 09:20:54
| 建立了一种基于单克隆抗体的直接竞争性酶联免疫吸附试验 (dcELISA)用于检测新的beta-肾上腺素能激活剂苯乙醇胺A (PEAA).在分别通过重氮反应和过碘酸钠法将其偶联到载体蛋白产生PEAA-BSA,PEAA-OVA和PEAA-HRP前，以 RANEY (R)镍为催化剂合成了一种PEAA的衍生物 PEAA-NH2。以PEAA-BSA为免疫原免疫3只Balb/C小鼠，获得了稳定产生对PEAA抗体的3个克隆。随后，用单克隆抗体(2H8)和PEAA-HRP建立了dcELISA方法。PEAA浓度在0.025-2.025ng/mL范围内，IC50值为0.204ng/mL。该单克隆抗体对PEAA及其衍生物PEAA-NH2高度特异，对莱克多巴胺有少量交叉反应(CR)(CR = 8.3%)，而对15种其它的beta-激活剂化合物的交叉反应可忽略(CR < 0.1%)。对样品测试，空白样品（猪肉、猪肝、猪尿和饲料）的LOD (检测限)值分别为0.431，0.455,0.225和8.693ng/mL；对应的LOQ（定量限）值分别0.578,0.647,0,360和14.477ng/mL。 回收率范围从82%到120%。批间和批内变异系数分别为13.33%和10.53%以下。此外dcELISA还用了液相色谱偶联质谱(LC-MS/MS)法评价，结果显示二法有高相关系数。总之，结果提示此 dcELISA以高灵敏度和特异性可用于可靠检测真实样品中PEAA的残留。 |
2楼2016-01-06 01:18:23