专一识别脱落酸甲酯的单克隆抗体的制备与应用

专一识别２－顺（Ｓ）ＡＢＡ甲酯的单克隆抗体来源于以ＡＢＡ分子中的１－ＣＯＯＨ为偶联位点合成的免疫原。它与游离态ＡＢＡ和结合态ＡＢＡ葡萄糖酯的交叉反应仅分别为１％与３．５％，而与ＡＢＡ类似物，如２－顺－黄质醛、紫黄质以及ＡＢＡ的２－反式异构体和（Ｒ）－对映体则无交叉反应。利用该抗体建立的高度灵敏和精确的ＡＢＡｍｅ酶联免疫测定法，其检测线性范围为０．０４８～１．５２ｐｍｏｌ。通过ＡＢＡｍｅＥＬＩＳＡ和ＧＡ１＋３ＥＬＩＳＡ分析可知羊蹄叶片衰老与内源ＧＡ１＋３／ＡＢＡ比值的下降有关。

Preparation and Application of Monoclonal Antibodies Specific for Abscisic Acid Methyl Ester

A monoclonal antibody 5 E ED7-51, IgG1 which specifically recognizes abscisic acid methyl ester(ABAme) is described. This antibodywas derived from an immunogen inwhich the carrier proteins were linked toABA at 1-COOH. It showed very littlecross-reactivity with ABA (1 %) orABA glucosyl ester (3. 5% ). Othercompounds structurally similar to ABA,such as 2-cis-xanthoxin, violaxanthinand both the 2, thans-isomer and the(R )-enantiomer of ABA, were notfound to exhibit any cross-reactivity(Table 1 ). Based on the use of this antibody, an enzyme--linked immunosorbent assay (ELISA) with high sensitivity and precision was developed, whichdisplayed a linear range of 0. 048 1. 52 pmol of ABAme (Fig. I ). Usingthis ELISA, as well as an establishedELISA for gibberellins (GAl 3 ), thelevels of ABA and GA, 3 in senescingleaves of Ruga Ppeicus were examined. The resultS showed that the decrease in GA1 3/ABA ratio was connected with the senescence of leaves ofvigu (Fig. 3).