%0 Journal Article %1 citeulike:469345 %A Ligler, F. S. %A Bredehorst, R. %A Talebian, A. %A Shriver, L. C. %A Hammer, C. F. %A Sheridan, J. P. %A Vogel, C. W. %A Gaber, B. P. %C Bio/Molecular Engineering Branch, Naval Research Laboratory, Washington, DC 20375-5000. %D 1987 %J Anal Biochem %K l-i-l-a competition complement immunoassay homogeneous ag_detection liposome %N 2 %P 369--375 %T A homogeneous immunoassay for the mycotoxin T-2 utilizing liposomes, monoclonal antibodies, and complement. %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=3661986 %V 163 %X The trichothecene mycotoxin T-2 is a fungal metabolite known to contaminate agricultural products and cause intoxication of humans and animals. We have developed a homogeneous competition inhibition assay for T-2 mycotoxin based on complement-mediated lysis of liposomes. The T-2 mycotoxin was converted to an acid chloride derivative, subsequently coupled to the amino group of phosphatidylethanolamine, and incorporated with the phospholipid into unilamellar liposomes. Carboxyfluorescein, which is self-quenched at high concentrations, was entrapped in the liposomes as a release marker. We used a monoclonal IgG1 antibody specific for T-2 mycotoxin and a polyclonal anti-mouse Ig as a secondary antibody since the anti-T-2 IgG1 does not activate complement. In the absence of free T-2, the liposomes were lysed within 30 min after the addition of complement, releasing carboxyfluorescein into the surrounding buffer. In the presence of free T-2 toxin, the binding of antibodies to the liposomes was reduced, causing a corresponding decrease in lysis. This assay proved to be sensitive to T-2 toxin levels as low as 2 ng, which is 10-fold more sensitive than the present enzyme immunoassay using the same antibodies.

@article{citeulike:469345, abstract = {The trichothecene mycotoxin T-2 is a fungal metabolite known to contaminate agricultural products and cause intoxication of humans and animals. We have developed a homogeneous competition inhibition assay for T-2 mycotoxin based on complement-mediated lysis of liposomes. The T-2 mycotoxin was converted to an acid chloride derivative, subsequently coupled to the amino group of phosphatidylethanolamine, and incorporated with the phospholipid into unilamellar liposomes. Carboxyfluorescein, which is self-quenched at high concentrations, was entrapped in the liposomes as a release marker. We used a monoclonal IgG1 antibody specific for T-2 mycotoxin and a polyclonal anti-mouse Ig as a secondary antibody since the anti-T-2 IgG1 does not activate complement. In the absence of free T-2, the liposomes were lysed within 30 min after the addition of complement, releasing carboxyfluorescein into the surrounding buffer. In the presence of free T-2 toxin, the binding of antibodies to the liposomes was reduced, causing a corresponding decrease in lysis. This assay proved to be sensitive to T-2 toxin levels as low as 2 ng, which is 10-fold more sensitive than the present enzyme immunoassay using the same antibodies.}, added-at = {2006-07-07T01:10:50.000+0200}, address = {Bio/Molecular Engineering Branch, Naval Research Laboratory, Washington, DC 20375-5000.}, author = {Ligler, F. S. and Bredehorst, R. and Talebian, A. and Shriver, L. C. and Hammer, C. F. and Sheridan, J. P. and Vogel, C. W. and Gaber, B. P.}, biburl = {https://www.bibsonomy.org/bibtex/27aff383a9567a661631ae2a026356517/biblio24}, citeulike-article-id = {469345}, interhash = {9e9ac3ba7c7aff468846ad1108d6fd85}, intrahash = {7aff383a9567a661631ae2a026356517}, issn = {0003-2697}, journal = {Anal Biochem}, keywords = {l-i-l-a competition complement immunoassay homogeneous ag_detection liposome}, month = {June}, number = 2, pages = {369--375}, priority = {2}, timestamp = {2006-07-07T01:10:50.000+0200}, title = {A homogeneous immunoassay for the mycotoxin T-2 utilizing liposomes, monoclonal antibodies, and complement.}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=3661986}, volume = 163, year = 1987 }