Nagasaki T, Yasuda S, Imai T
Laboratory of Biomass Resources Utilization, Graduate School of Bioagricultural Sciences, Nagoya University, 464-8601, Nagoya, Japan.
Phytochemistry. 2001 Nov;58(6):833-40. doi: 10.1016/s0031-9422(01)00323-5.
In order to immunolabel heartwood extractives in Japanese cedar (Sugi, Cryptomeria japonica), we attempted to prepare antibodies against agatharesinol, a major norlignan of these heartwood extractives. Agatharesinol by itself is not antigenic due to its low-M(r), and thus was covalently bound to bovine serum albumin in order to synthesize an antigenic hapten-carrier conjugate (artificial antigen). The number of agatharesinol molecules per artificial antigen molecule was estimated as 27-28 by quantifying Lys in an acid hydrolysate of the artificial antigen by HPLC. Reaction between the artificial antigen and serum obtained from a rabbit immunized with the artificial antigen was competitively inhibited by agatharesinol, indicating the successful production of anti-agatharesinol antibodies. Inhibition by sequirin C, another major norlignan in Sugi, was weaker than that by agatharesinol. Furthermore, an EtOAc soluble fraction, which contains mainly norlignans, inhibited the reaction more strongly than any of the other fractions of Sugi heartwood extractives. Thus, the antiserum we have produced reacts most strongly with agatharesinol and recognizes norlignans almost selectively among Sugi heartwood extractives.
为了对日本柳杉（杉木，Cryptomeria japonica）的心材提取物进行免疫标记，我们尝试制备针对阿加他雷醇的抗体，阿加他雷醇是这些心材提取物中的一种主要去甲木脂素。由于阿加他雷醇分子量较低，其本身没有抗原性，因此将其与牛血清白蛋白共价结合，以合成一种抗原性半抗原 - 载体偶联物（人工抗原）。通过高效液相色谱法对人工抗原的酸水解产物中的赖氨酸进行定量分析，估计每个人工抗原分子中的阿加他雷醇分子数为27 - 28个。人工抗原与用该人工抗原免疫的兔子血清之间的反应受到阿加他雷醇的竞争性抑制，这表明成功制备了抗阿加他雷醇抗体。杉木中另一种主要去甲木脂素水苏素C的抑制作用比阿加他雷醇弱。此外，主要含有去甲木脂素的乙酸乙酯可溶部分比杉木心材提取物的任何其他部分对反应的抑制作用更强。因此，我们制备的抗血清与阿加他雷醇反应最强，并且在杉木心材提取物中几乎选择性地识别去甲木脂素。
