Immobilization of small molecules on solid matrices: A novel approach to enzyme-linked immunosorbent assay screening for saxitoxin and evaluation of anti-saxitoxin antibodies

摘要: A novel enzyme-linked immunosorbent assay (ELISA) technology was developed for detecting saxitoxin or evaluation of anti-saxitoxin antibodies, which is based on non-covalent immobilization "free' to Maxisorp microtitre plates. The effect pH studied in media with wide-range buffering capacities (piperazine-glycylglycine and barbiturate buffers). Increasing resulted better responses, although this mainly due non-specific interactions. At 10.0, however, quite effective specific. same pattern found under four different conditions; absence vs presence bovine serum albumin precoating 150 mM NaCl. best results (high specific response) were achieved the method choice involved 5 micrograms/ml followed by addition microM 0.01 M piperazine-glycylglycine buffer, 10.0. efficacy demonstrated a polyclonal rabbit antibody compared conventional ELISA using saxitoxin-bovine conjugate as coating antigen. In experiments investigating cross-reactivities various derivatives competitive assay, significantly greater sensitivity approach, e.g. 35 pM could be detected (3 x 10(4) times lower concentrations than conjugate). works well mussel tissue homogenates, because it does not require use covalent saxitoxin-carrier conjugates offers simpler alternative traditional saxitoxin.