Hradecká Veronika, Novák Ondrej, Havlícek Libor, Strnad Miroslav
Laboratory of Growth Regulators, Palacký University & Institute of Experimental Botany ASCR, Slechtitelů 11, CZ-783 71 Olomouc, Czech Republic.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Mar 1;847(2):162-73. doi: 10.1016/j.jchromb.2006.09.034. Epub 2006 Oct 24.
Polyclonal antibodies with high specificity for C1-immobilised (+)-cis,trans-abscisic acid (ABA) were raised, characterised by enzyme-linked immunosorbent assay (ELISA) and used for preparation of an immunoaffinity chromatography (IAC) gel. The detection limit of the ELISA was approximately 4.6x10(-10)mol/L. Sensitive electrospray liquid chromatography-mass spectrometry (LC-ESI-MS) methods were also developed with detection limits below 0.1x10(-12)mol. The IAC allowed quick, single-step processing of samples prior to the analyses. The LC-ESI-MS and LC-ELISA techniques were used for comparative estimation of endogenous ABA levels in immunoaffinity purified extracts of normal and water-stressed Nicotiana tabacum L. leaves. The analytical approaches were validated using deuterium- and tritium-labelled internal standards, respectively. The IAC method was found to be highly effective, sensitive and convenient for isolating the target analyte from plant material.
制备了对固定有C1的（+）-顺式，反式脱落酸（ABA）具有高特异性的多克隆抗体，通过酶联免疫吸附测定（ELISA）对其进行表征，并用于制备免疫亲和色谱（IAC）凝胶。ELISA的检测限约为4.6×10（-10）mol/L。还开发了灵敏的电喷雾液相色谱-质谱联用（LC-ESI-MS）方法，检测限低于0.1×10（-12）mol。IAC允许在分析前对样品进行快速的单步处理。LC-ESI-MS和LC-ELISA技术用于比较估计正常和水分胁迫的烟草叶片免疫亲和纯化提取物中的内源ABA水平。分别使用氘和氚标记的内标对分析方法进行了验证。发现IAC方法对于从植物材料中分离目标分析物非常有效、灵敏且方便。
