Cells Useful for Immuno-Based Botulinum Toxin Serotype A Activity Assays

Abstract

The present specification discloses clonal cell lines susceptible to BoNT/A intoxication, methods of producing such clonal cell lines, and methods of detecting Botulinum toxin serotype A activity using such clonal cell lines.

We claim: 
     
         1 . An established single cell derived clonal cell line comprising cells susceptible to intoxication by botulinum toxin type A (BoNT/A),
 wherein the cells exhibit at least a 1.5-fold increase in mRNA expression levels after differentiation in the genes ADAMTS9, ATAD2, CDC45L, CNTN1, Cyclin A, Cyclin E, E2F1, E2F2, E2F7, FGFR2, MYBL1, MYBL2, NDC80, NDN, ORC1L, PLS3, RAD54L, RBL1, RRM2, S1PR3, SPC24, SPC25, TFDP1, TFPI2, TK1, TTK, TYMS, TYK and ZWINT relative to the mRNA expression levels of each of these genes in cells from the parental SiMa cell line DSMZ ACC 164; and,   wherein the cells exhibit at least a 1.5-fold decrease in mRNA expression levels after differentiation in the genes CDKN2A and CNTN2 as compared to the mRNA expression levels of each of these genes in cells from the parental SiMa cell line DSMZ ACC 164; and,   wherein the clonal cell line comprises cells susceptible to BoNT/A intoxication by about 100 pM or less of a BoNT/A.   
     
     
         2 . The established clonal cell line of  claim 1 , wherein the cells exhibit at least a 2.0-fold difference in gene expression levels of at least one gene as compared to the expression levels of the gene in cells from the parental SiMa cell line DSMZ ACC 164. 
     
     
         3 . The established clonal cell line of  claim 1 , wherein the cells exhibit at least a 3.0-fold difference in gene expression levels of at least one gene as compared to the expression levels of the gene in cells from the parental SiMa cell line DSMZ ACC 164. 
     
     
         4 . The established clonal cell line of  claim 1 , wherein the cells exhibit at least a 4.0-fold difference in gene expression levels of at least one gene as compared to the expression levels of the gene in cells from the parental SiMa cell line DSMZ ACC 164. 
     
     
         5 . The established clonal cell line of  claim 1 , wherein the cell from an established clonal cell line exhibits about 100 pM or less of a BoNT/A for at least 5 or more cell passages.