M30 Apoptosense® ELISA is an enzyme

Enzyme

Enzymes are proteins that catalyze chemical reactions. In enzymatic reactions, the molecules at the beginning of the process, called substrates, are converted into different molecules, called products. Almost all chemical reactions in a biological cell need enzymes in order to occur at rates...

-linked immunosorbent assay developed for the detection of soluble caspase-cleaved keratin 18

Keratin 18

Keratin 18 is a type I cytokeratin. It is, together with its filament partner keratin 8, perhaps the most commonly found products of the intermediate filament gene family. They are expressed in single layer epithelial tissues of the body. Mutations in this gene have been linked to cryptogenic...

 (ccK18, K18-Asp396, formerly cytokeratin 18, ccCK18 or CK18-Asp396).

History

The M30 Apoptosense® ELISA was developed by Peviva AB (Bromma, Sweden) in collaboration with the Karolinska Institute in 2000.

Uses

The M30 Apoptosense® ELISA is a non-invasive test for the detection of apoptosis of epithelially derived cells. Caspases are activated in apoptotic cells and cleave intracellular protein substrates. K18 is one such substrate, expressed by many epithelial cells (e.g. hepatocytes, intestinal cells). Cleaved K18 is released into the circulation after cell death. The main uses of M30 Apoptosense® ELISA are to:

1. Measure apoptosis of tumor cells in response to cytotoxic drugs (breast, prostate and other forms of carcinomas) .
2. Measure hepatocyte apoptosis to aid the diagnosis of non-alcoholic steatohepatitis (NASH) and steatosis in patients with hepatitis C virus infection .
3. Serve as a biomarkers of epithelial apoptosis in liver and intestinal graft-versus-host disease .

Biology

Caspases cleave keratin 18 at two sites during apoptosis. Cleavage at Asp396 generates a neo-epitope recognized by the monoclonal antibody M30 . This antibody does not recognize uncleaved K18 and is therefore specific for apoptotic epithelial cells. M30 Apoptosense® ELISA utilizes a second monoclonal antibody (M5) which recognizes an epitope N-terminal from the M30 epitope.

M30 Apoptosense® ELISA can be combined with the M65® ELISA (Peviva AB) to determine cell death mode (apoptosis versus necrosis) .