M13 Bacteriophages as Bioreceptors in Biosensor Device

摘要

New recognition probes sensible, specific and robust is one of the major problems of biosensor assay. Detection biosensors has utilized antibodies or enzymes as bioreceptors; however, these have numerous disadvantages of limited binding sites and physico-chemical instabilities, can negatively affect capture and detection of target in diagnostic device. In this contest, Phage-Display provides a valuable technique for obtaining large amounts of specific and robustness bio-probes in a relatively short time. This technique relies on the ability of M13 bacteriophages (or phages) to display specific and selective target-binding peptides on major coat protein pⅧ of their surface. In this work, we used P9b phage clone, displaying a foreign peptide QRKLAAKLT to selectively recognize Pseudomonas aeruginosa like bioreceptor. We describe different methods of functionalization to realize a selective bacteria biosensor surfaces. Several surfaces, such as latex and magnetic beads and polymeric surfaces such as mica, APTES and PEI, were functionalized by covalent bonds or physisorption with P9b. The efficiency of the surface functionalization procedures was evaluated by ELISA and AFM, while capture efficiency of the anchored phages has been assessed by plate count and Fluorescence microscopy. The results of this work pave the way to the use of phages as bioreceptor.