Cytotoxicity assay in which the cell toxicity is assayed using different kinds of techniques. Some of these techniques patented are given below.
Patent no: 6,982,152 titled ‘Cytotoxicity Assay’ with the abstract: Disclosed are a method and a corresponding kit for determining the cytotoxicity of a test agent. The method includes the steps of adding a pre-determined amount of the test agent to a vessel containing living cells in culture medium. The cells are then incubated for a pre-determined amount of time. To the cells and culture medium is then added a reagent mixture that is non-toxic to the living cells. In the preferred embodiment, the reagent mixture contains a solvent, a dye, an electron transfer agent, a substrate for a cytoplasmic enzyme having a half-life greater than two hours and NAD+. The contents of the vessel are then measured for production of the reduced state of the dye, the production of the reduced state of the dye being caused by a cytoplasmic enzyme specific for the substrate in the reagent mixture. In the preferred embodiment, the production of the reduced state of the dye is caused by lactate dehydrogenase released from dead and dying cells within the vessel. The kits contain components necessary for performing the method.
Patent no: 5,514,708 titled ‘Cytotoxic metabolites from Myriapora truncata’ with the abstract : The present invention is based upon the discovery that the methanol extract of the bryozoan Myriapora truncata showed potent cytotoxicity against L1210 murine leukemia cells (99% inhibition at 50 .mu.g/mL). Fractionation and purification of active components from this extract, guided by a cytotoxicity assay, resulted in the isolation of a novel, highly cytotoxic polyketide-derived metabolite MT-332 (Compound 3) and its equilibrium isomer (Compound 4), along with two less active compounds, MT-381 (Compound 1) and MT-381-B (Compound 2). The equilibrium mixture of Compounds 3 and 4 showed 88% inhibition at 0.2 .mu.g/mL against L1210 cels. ##STR1##
Patent no: 5,306,624 titled Process of quantifying cell number with the abstract: The present invention provides a process of quantifying the number of viable cells in an aqueous suspension of cells using an energy-emitting non-hazardous probe and a probe-trigger. The process provides quantification data in short periods of time without the use of hazardous materials. A process of the present invention can also be used to quantify negatively charged particle number, assay for cytotoxicity, assay for cell proliferation and assay for cell differentiation. Still further, the present invention provides an assay kit for quantification of cells or negatively charged particles.
Patent no: 5,055,400 titled ‘Leukotoxin gene of pasteurella haemolytica’ with the abstract: The gene coding for Pasteurella haemolytica leukotoxin can be cloned in a plasmic expressed in Escherichia coli. The leukotoxin gene can be isolated from a clone bank of P. haemolytica. The clone bank is constructed by partial digestion of genomic DNA. The resultant 5 to 10 kilobase-pair fragments are ligated into plasmid vector pBR322. The resultant clones are screened for the production of P. haemolytica soluble antigens by a colony enzyme-linked immunosorbent assay blot method with a rabbit antiserum raised against the soluble antigens. The clones producing P. haemolytica soluble antigens are then analyzed for the production of the leukotoxin by a cytotoxicity assay with cells from a bovine leukemia-derived B-lymphocyte cell line as the target cells. Positive clones are identified, and subsequent restriction analysis of the recombinant plasmids shows the same insert DNA is cloned in the plasmid vector. The DNA sequence analysis of the insert DNA reveals regions coding for the leukotoxin.
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