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首页> 外文期刊>Acta Horticulturae >Preliminary data on an antibody for an isoxanthohumol ELISA.
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Preliminary data on an antibody for an isoxanthohumol ELISA.

机译：异黄腐酚ELISA抗体的初步数据。

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摘要

Isoxanthohumol is a weak estrogenic prenylflavanone from hop that possesses anti-carcinogenic effects. It is also a precursor of 8-Prenylnaringenin. A carboxylic acid hapten of prenylflavanone was synthesized with a spacer arm of 2 carbons at the C4' position. The hapten was characterized by 1H-NMR, 13C-NMR, IR, MS (ESI+) and HR-MS (ESI+). It was coupled to Bovine Serum Albumin (BSA) by the activated ester method. The coupling efficiency was assessed by MALDI-MS. It was found that the average coupling ratio was 26.5+or-1. Two antibodies were raised in two rabbits. For the preliminary tests, competitive ELISA was chosen as the testing method. To undertake such tests, H-prenylflavanone was coupled to swine thyroglobulin with a 50:1 ratio in order to perform the coating step. Antibodies were tested for titres and the best was retained for further investigations. A first standard curve was obtained with the following conditions: coating 0.03 micro g/ml, Ac1: 1/250,000; Ac2: 1/10,000. Specificity tests were undertaken using various flavones and flavanones. Cross-reactions were: Xanthohumol: 0.09%, 8-Prenylnaringenin: 2.71%, Naringenin: 12.6%, Liquiritigenin: 3.2%, Eriodictyol 1.6%. A sample of LifenolReg. was assayed after extraction with ethanol:water 75:25 or enzyme hydrolysis and extraction with ethyl-acetate. The average concentration was 6.3+or-2.0% or 6.0+or-2.1% after one or the other extraction.
机译：异黄腐酚是一种来自啤酒花的弱雌激素异戊烯基黄酮，具有抗癌作用。它也是8-Prenylnaringenin的前体。合成了异戊烯基黄酮的羧酸半抗原，其在C4'位置具有2个碳的间隔臂。半抗原的特征是 1 H-NMR， 13 C-NMR，IR，MS（ESI +）和HR-MS（ESI +）。通过活化酯法将其与牛血清白蛋白（BSA）偶联。通过MALDI-MS评估偶联效率。发现平均偶合比为26.5+或-1。在两只兔子中产生了两种抗体。对于初步测试，选择竞争性ELISA作为测试方法。为了进行这种测试，将H-异戊烯基黄酮与猪甲状腺球蛋白以50：1的比例偶联，以进行包衣步骤。测试抗体的滴度，并保留最佳抗体以进行进一步研究。在以下条件下获得第一标准曲线：包衣0.03微克/毫升，Ac1：1 / 250,000; Ac2：1 / 10,000。使用各种黄酮和黄烷酮进行了特异性测试。交叉反应为：黄腐酚：0.09％，8-苯甲基柚皮苷：2.71％，柚皮苷：126.6％，皂苷元：3.2％，松香醇1.6％。 LifenolReg的样本。用乙醇：水75∶25萃取或酶水解并用乙酸乙酯萃取后，测定糖含量。一种或另一种萃取后的平均浓度为6.3+或-2.0％或6.0 +或-2.1％。

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