Inventors:
Hanping Feng - Framingham MA, US
Saul Tzipori - Shrewsbury MA, US
Guilin Yang - Shenzhen, CN
Assignee:
Tufts University - Boston MA
International Classification:
G01N 33/53
US Classification:
435 732, 435 72, 435 71, 435 29, 435 34, 435 39, 4242471, 4241671
Abstract:
Cell-based methods for rapid real time assay of a presence of toxin and/or cells are provided, using an assay having a toxin-enhancing antibody and a sensitive cell line carrying FcyR receptors, and kits for this assay. An ultrasensitive cell based immunocytotoxicity assay for detecting less then 1 pg/ml of toxins in clinical samples. A TcdA-specific monoclonal antibody, AIH3, was found to significantly enhance the cytotoxicity of TcdA to macrophages and monocytes. The AIH3-dependent enhancement of glucosyltransferase activity, cytoskeleton disruption, and TNF-a production induced by TcdA was demonstrated also in RAW 264. 7 cells. Methods for high level recombinant expression of toxins in cells, and vectors for expression, strains of carrying the vectors are provided.