Journal of
Medical Genetics and Genomics

  • Abbreviation: J. Med. Genet. Genomics
  • Language: English
  • ISSN: 2141-2278
  • DOI: 10.5897/JMGG
  • Start Year: 2009
  • Published Articles: 48

Full Length Research Paper

Preparation and partial characterization of monoclonal antibody against human phosphdiesterase 5 (PDE5)

Fawzi Alsheyab1, Nizar Abuharfil1, Mohanad Aldarouish1, Dalal Awad2 and Yousef Mosameh3*
1Department of Biotechnology, Jordan University of Science and Technology (JUST), Irbid- Jordan. 2Department of Biological Sciences, Yarmouk University, Irbid-Jordan. 3Renad Medical Laboratory, Amman-Jordan.
Email: [email protected]

  • Article Number - CBC0F609054
  • Vol.5(1), pp. 6 - 13, February 2013
  •  Accepted: 01 June 2012
  •  Published: 28 February 2013

Abstract

Phosphodiesterase 5 (PDE5) is a catalytic enzyme for degradation of cyclic guanosine monophosphate (cGMP) in human smooth muscle cells. Inhibition of this enzyme by certain chemicals including sildenafil, vardenafil and tadalafil, is considered a breakthrough treatment of erectile dysfunction. Unfortunately, these drugs are associated with several side effects. In this study, another strategy is adopted in the inhibition of the PDE5 enzyme by using a monoclonal antibody (Mab) against the enzyme. The Mab was prepared by immunizing a mouse, taking the spleen to make single cell suspension of lymphocytes. Hybridoma cells were obtained by fusion of BALB/c myeloma cells with the immunized lymphocytes using the polyethylene glycol method. Several clones were produced that have been separated by limiting dilution method. The antibodies were tested by enzyme-linked immunosorbent assay (ELISA) and western blot. The obtained Mab has titer of 625, immunoglobulin G1 (IgG1) isotype and 3.9 × 104 mol-1 affinity constant.

 

Key words: Phosphodiesterase (PDE), chemical inhibition, monoclonal antibody (Mab).

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