Development of rapid and reliable immunochemical methods for monitoring psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine; Pyb) and psilocin (dephosporylated metabolite; Psi), the psychoactive compounds contained within hallucinogenic mushrooms (magic mushrooms), is desirable in order to identify these mushrooms and regulate their illicit use. Because no antibody was publicly available for this purpose, we generated two independent monoclonal antibodies (mAbs) against Pyb or Psi, and then developed enzyme-linked immunosorbent assays (ELISAs) by using them. To generate the specific antibodies, novel immunogenic conjugates were prepared by linking Pyb or Psi molecules to carrier proteins by modifying their 2-(N,N-dimethylamino)ethyl side chains. Spleen cells from mice immunized with these conjugates were fused with P3/NS1/1-Ag4-1 myeloma cells, and hybridoma clones secreting anti-Pyb and anti-Psi mAbs were established. These mAbs were characterized for their biochemical features and then applied to competitive ELISAs, which used microplates coated with Pyb or Psi linked with albumin. These ELISAs enabled the determination of Pyb or Psi with measurable ranges of ca. 0.20−20 or 0.040−2.0 μg/assay (limit of detection was 0.14 or 0.029 μg/assay), respectively. The related tryptamines were satisfactorily discriminated as exemplified by the cross-reactivity of the ELISA to determine Pyb (or Psi) with Psi (or Pyb) that were found to be 2.8% (or <0.5%), respectively. The Pyb and Psi contents in a dried powder of the hallucinogenic mushroom, Psilocybe cubensis, were determined to be 0.39 and 0.32 (w/w)%, respectively. The ELISAs developed using the current mAbs are promising tools for identifying illegal hallucinogenic mushrooms.

为了确定致幻蘑菇（魔术蘑菇）中包含的精神活性化合物，需要开发一种快速，可靠的免疫化学方法来监测psilocybin（4-磷酸酰氧基-N，N-二甲基色胺； Pyb）和psilocin（脱磷代谢物； Psi）。这些蘑菇并规范其非法使用。由于没有公开的抗体可用于此目的，因此我们生成了两种针对Pyb或Psi的独立单克隆抗体（mAb），然后通过使用它们开发了酶联免疫吸附测定（ELISA）。为了产生特异性抗体，通过修饰Pyb或Psi分子的2-（N，N-二甲基氨基）乙基侧链。将用这些缀合物免疫的小鼠的脾细胞与P3 / NS1 / 1-Ag4-1骨髓瘤细胞融合，并建立分泌抗Pyb和抗Psi mAb的杂交瘤克隆。对这些单克隆抗体的生化特征进行表征，然后将其应用于竞争性ELISA中，该ELISA使用涂有Pyb或与白蛋白连接的Psi的微孔板。这些ELISA可以测定Pyb或Psi的可测范围为ca。分别为0.20-20或0.040-2.0μg/测定（检出限为0.14或0.029μg/测定）。ELISA的交叉反应性可以令人满意地区分相关的色胺，以确定Pyb（或Psi）与Psi（或Pyb）的比率分别为2.8％（或<0.5％）。在致幻蘑菇，的干燥粉末的PYB和帕普西内容裸盖菇霜霉病，被确定为0.39和0.32（重量/重量）％之间。使用当前的单克隆抗体开发的ELISA是用于鉴定非法致幻蘑菇的有前途的工具。