Nishi Kosuke, Ishiuchi Masatake, Morimune Kosuke, Ohkawa Hideo
Research Center for Environmental Genomics, Kobe University, Japan.
J Agric Food Chem. 2005 Jun 29;53(13):5096-104. doi: 10.1021/jf050246t.
A monoclonal antibody (mab) selective for the thiomethyl-s-triazine herbicide simetryn was obtained and characterized in enzyme-linked immunosorbent assay (ELISA). An IC(50) value for simetryn was 8.5 ng/mL, and the detection range extended from 1.1 to 70 ng/mL in ELISA. The cDNAs encoding variable heavy chain (VH) and variable light chain (VL) of the mab were cloned to produce various recombinant antibodies. Single-chain variable fragment (scFv) antibodies derived from the mab were characterized in ELISA and showed similar reactivities and specificities to the parent mab. A urea denaturation test revealed that the scFv antibodies bound to simetryn were more stable than those in the absence of antigen. A sandwich ELISA based on VH and VL fragments of the mab was successfully developed and showed similar sensitivity to those based on the mab and scFv antibodies in ELISA. In the recovery experiments using spiked environmental samples, the results obtained in ELISA based on the mab were favorably correlated with those by HPLC.
获得了一种对硫甲基 - s - 三嗪除草剂西草净具有选择性的单克隆抗体（mab），并在酶联免疫吸附测定（ELISA）中对其进行了表征。西草净的半数抑制浓度（IC50）值为8.5 ng/mL，在ELISA中的检测范围为1.1至70 ng/mL。克隆了编码该mab重链可变区（VH）和轻链可变区（VL）的cDNA，以产生各种重组抗体。源自该mab的单链可变片段（scFv）抗体在ELISA中进行了表征，显示出与亲本mab相似的反应性和特异性。尿素变性试验表明，与西草净结合的scFv抗体比未结合抗原的抗体更稳定。基于该mab的VH和VL片段成功开发了一种夹心ELISA，其在ELISA中的灵敏度与基于mab和scFv抗体的ELISA相似。在使用加标环境样品的回收率实验中，基于mab的ELISA结果与高效液相色谱法（HPLC）的结果具有良好的相关性。
