A MONOCLONAL-ANTIBODY TO (S)-ABSCISIC ACID - ITS CHARACTERIZATION AND USE IN A RADIOIMMUNOASSAY FOR MEASURING ABSCISIC-ACID IN CRUDE EXTRACTS OF CEREAL AND LUPIN LEAVES
A MONOCLONAL-ANTIBODY TO (S)-ABSCISIC ACID - ITS CHARACTERIZATION AND USE IN A RADIOIMMUNOASSAY FOR MEASURING ABSCISIC-ACID IN CRUDE EXTRACTS OF CEREAL AND LUPIN LEAVES
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DOI：
10.1007/bf00401020
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发表时间：
1988-03-01
期刊：
影响因子：
4.3
通讯作者：
LOVEYS, BR
中科院分区：
文献类型：
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作者：
QUARRIE, SA;WHITFORD, PN;LOVEYS, BR
一种产生脱落酸（ABA）的单克隆抗体已被表征，并描述了使用该抗体的ABA放射免疫测定（RIA）的发展。该抗体对(S)-cis，trans-ABA的游离酸具有较高的选择性。使用该抗体，可以在RIA中可靠地测定ABA，测定范围为100至4000 pg(0.4至15 pmol) /瓶。由于甲醇和丙酮影响ABA-抗体结合，因此用水提取ABA。水提取ABA的效果与甲醇和丙酮同等。小麦（Triticum aestivum）、玉米（Zea mays）和卢平（Lupinus angustifolius， L. cosentinii）叶片的粗水提物可以在不受其他免疫反应物质严重干扰的情况下进行分析。这可以通过薄层色谱（TLC）和高效液相色谱（HPLC）分离粗提物的免疫反应性分布，或通过与理化分析方法的比较来证明。用RIA和TLC纯化后的电子捕获检测器气相色谱或HPLC纯化后的气相色谱-质谱联用（GC-MS）对粗提取物进行分析，发现初始叶样品中的ABA浓度非常相似。然而，RIA分析的豌豆种子的粗水提取物导致相当大的高估ABA存在的量。采用高效液相色谱法纯化豌豆籽提取物后，采用气相色谱-质谱法和RIA法测定ABA含量的结果相似。虽然RIA不能用于分析豌豆种子粗提取物中的ABA，但很可能可以成功地分析其他几种植物叶片的粗提取物。
A monoclonal antibody produced to abscisic acid (ABA) has been characterised and the development of a radioimmunoassay (RIA) for ABA using the antibody is described. The antibody had a high selectivity for the free acid of (S)-cis,trans-ABA. Using the antibody, ABA could be assayed reliably in the RIA over a range from 100 to 4000 pg (0.4 to 15 pmol) ABA per assay vial. As methanol and acetone affected ABA-antibody binding, water was used to extract ABA from leaves. Water was as effective as aqueous methanol and acetone in extracting the ABA present. Crude aqueous extracts of wheat [Triticum aestivum], maize [Zea mays] and lupin [Lupinus angustifolius, L. cosentinii] leaves could be analysed without serious interference from other immunoreactive material. This was shown by measuring the distribution of immunoreactivity in crude extracts separated by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), or by comparing the assay with physicochemical methods of analysis. Analysis of crude extracts by RIA and either, after TLC purification, by gas chromatography using an electron-capture detector or, after HPLC purification, by combined gas chromatography-mass spectrometry (GC-MS) gave very similar ABA concentrations in the initial leaf samples. However, RIA analysis of crude aqueous extract of pea seeds resulted in considerable overestimation of the amount of ABA present. Determinations of ABA content by GC-MS and RIA were similar after pea seed extracts had been purified by HPLC. Although the RIA could not be used to analyse ABA in crude extracts of pea seeds, it is likely that crude extracts of leaves of several other species may be assayed successfully.