AI Article Synopsis
- A mouse monoclonal antibody (MAb) developed through hybridoma technology targets a 150-kD surface antigen of Entamoeba histolytica, effectively blocking the microorganism's ability to adhere and cause damage to mammalian cells.
- The genes for the antibody's light chain and part of the heavy chain were cloned into Escherichia coli for expression, using a plasmid designed specifically for producing Fab fragments with a hexa-histidine tag.
- The resulting recombinant Fab fragments matched the specificity of the original MAb and notably inhibited the adherence of E. histolytica to red blood cells, demonstrating the efficiency of E. coli as an expression system for producing neutralizing antibodies.
Article Abstract
A mouse monoclonal antibody (MAb) (EH3015, IgG1 with a K light chain) prepared by hybridoma technology recognizes a 150-kD surface antigen of Entamoeba histolytica and inhibits adherence and cytotoxicity of the ameba to mammalian cells. The genes encoding the light chain and the Fd region of the heavy chain of the MAb were cloned and expressed in Escherichia coli. The plasmid used was designed for the expression of Fab with a hexa-histidine tag in the periplasmic space. Recombinant Fab fragments were purified and analyzed by an indirect immunofluorescence antibody test and Western immunoblot. The specificity of the recombinant Fab fragment was comparable with the parent whole IgG. In addition, the Fab fragments significantly inhibited the adherence of E. histolytica to erythrocytes. These results suggest that the production of a neutralizing MAb in Escherichia coli is practical and efficient with this expression system.
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| http://dx.doi.org/10.4269/ajtmh.1999.60.1.9988319 | DOI Listing |
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