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Asian Journal of Scientific Research
Year: 2014  |  Volume: 7  |  Issue: 3  |  Page No.: 303 - 311

Lotus-leaf Inspired Hydrophobic Nanocomposite Matrices for Electrophoretic Separation of Bacterial Outer Membrane Proteins

Lourdusamy Arul Antony Maria, Vidya N. Chamundeswari and Meera Parthasarathy    

Abstract: Electrophoretic separation of membrane proteins is limited mainly by their intrinsic hydrophobicity and poor solubility arising from association with membrane lipid components. In this investigation, polyacrylamide gel matrix is made hydrophobic by introducing one-dimensional and two-dimensional carbon nanomaterials viz., multi-walled carbon nanotubes and graphene. The nanocomposite matrices were optimized for SDS-PAGE with water-soluble molecular weight marker proteins. SDS-PAGE in the nanocomposite matrices of a commercial outer membrane porin protein, OmpA indicated a significant decrease in anomalous migration of the protein with increasing carbon nanotube loading in the gel matrix. Outer membrane proteins of Escherichia coli, were isolated and characterized by Fourier Transform Infrared Spectroscopy. When the nanocomposite gels were tested for electrophoretic separation of outer membrane proteins isolated from E. coli, the resolution of protein bands improved with respect to the pristine polyacrylamide gel. Especially, graphene/polyacrylamide composite hydrogels yielded far better resolution and faster migration of the E. coli membrane proteins compared to multi-walled carbon nanotube/polyacrylamide composite hydrogels. Based on contact angle measurements of the composite hydrogels, the improved resolution of membrane proteins is attributed to the more hydrophobic environment rendered by carbon nanotubes and graphene. The present results could be useful to develop more hydrophobic nanocomposite gels exclusively for electrophoretic separation of membrane proteins isolated from different sources.

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