Development of Monoclonal Antibody-based Enzyme-linked Immunosorbent Assay for Detecting Tetrodotoxin
This study aimed to develop a method for the detection of tetrodotoxin (TTX) based on monoclonal antibody (MeAb).The hapten TTX was linked to carrier protein keyhole limpet hemocyanin (KLH) as immunogen,and linked to ovalbumin (OVA) as coating antigen by the Mannich method.Then the 6~8 weeks Babl/c mice were immunized.After cell amalgamation,a cell line with high specificity and sensitivity was obtained,and McAb was produced.The indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed for detecting TTX.The optimized working conditions of the icELISA were 4.0 μg/mL coating antigen,0.75 μg/mL McAb,45 min competitive time,at room temperature (20~25 ℃).The IC50 value of this method was 24.0 ng/mL,the working ranges were 5.2~107.6 ng/mL,the intra-assay and inter-assay coefficient of variation (CV %) were 4.2 and 4.5,respectively.This investigation will benefit the assay kit development for detecting TTX.
tetrodotoxin monoclonal antibody E LISA
ZHONG Qing Ping LIU Yan Ting SUN Yuan Ming YU Hoi Fu LEI Hong Tao
Guangdong provincial key laboratory of food quality and safety,South China Agricultural University,G Department of Applied Biology & Chemical Technology,Polytechnic University,Hong Kong,China
国际会议
太原
英文
220-224
2012-01-13（万方平台首次上网日期，不代表论文的发表时间）