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  • Facts To Know About Cytotoxicity Testing

    Johnson

    July 18, 2022
    Health
    Facts To Know About Cytotoxicity Testing

    Cytotoxicity assays are used to assess the effect of certain drug compounds on the human body. The method proves valuable in determining whether a certain drug candidate is responsible for nonaccidental cell destruction (referred to as necrosis) or programmed cell death (referred to as apoptosis). Cytotoxic compounds can hinder or stop cell proliferation, making them strong candidates for cancer treatment drugs. Cytotoxicity assays are performed to quantify the toxicity of new drug compounds. They help identify and understand the impact of a compound on the normal cells or specific types of cells. Cytotoxicity assays allow researchers to observe the effects of drugs on normal human cells as well as cells that have been intentionally damaged through radiation or drugs. Cytotoxicity assays also identify compounds that produce irreversible damage to cells, which could cause adverse side effects during clinical trials.

    A cytotoxicity testing reveals the extent to which a drug candidate is damaging to cells. Certain compounds can even induce apoptosis, or programmed cell death, while others trigger necrosis. In either case, dose-dependent assessment helps researchers make important decisions early in the drug discovery process. Cytotoxicity assays are developed to analyze the ability of certain drug compounds to destroy the healthy cells of an organism. Cytotoxic compounds can lead to accidental cell death (referred to as necrosis) or programmed cell death (referred to as apoptosis). During the nonclinical and clinical research phases of drug discovery and development, cytotoxicity assays assess the safety profile of the drug candidate. The method proves valuable in identifying off-target effects of certain drug compounds on the human body.

    In this method of ELISA, the analyte whose concentration is to be measured is sandwiched in-between two antibodies that bind to different regions (epitopes) on the antigen. These antibodies are referred to as detection antibody and capture antibody. In this assay, the capture antibody is coated to a microtiter plate in a 96-well format. The antigen binds to the capture antibody, and a detection antibody is used to measure the analyte. The antibody is conjugated to an enzyme, typically horseradish peroxidase (HRP), and detected via catalysis of a substrate which yields a colored product. Spectrophotometry is used to monitor the colored product, and a standard curve is utilized for calculating the antigen concentration in the sample.

    ELISA is a qualitative or quantitative assay based on the immune system. ELISA is used to detect antibodies in the blood and other body fluids. The type of elisa lab test kit you have will determine how you should use it. To run an ELISA on blood, blood must be serum and not plasma. The two separate instructions come with your kit from the manufacturer and should be followed carefully.

    ELISA is a common laboratory test used to detect the presence or quantify an analyte in a sample. It is also referred to as a “sandwich” type of assay as it requires both antibodies, one attached to the microtiter plate and one in solution, that bind to different regions of the targeted antigen. This sandwich is incubated with the protein of interest in samples from patients with or without disease. Protein conjugated with enzyme indicators, such as HRP or AP (alkaline phosphatase), are used for detection by their color change at appropriate pH and are measured using spectrophotometry.

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