Premium
Reusable linking chemistry for His‐6 tagged proteins in an affinity‐based porous silicon biosensor
Author(s) -
Bonanno Lisa,
DeLouise Lisa
Publication year - 2009
Publication title -
physica status solidi (a)
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.532
H-Index - 104
eISSN - 1862-6319
pISSN - 1862-6300
DOI - 10.1002/pssa.200881088
Subject(s) - biosensor , porous silicon , nanotechnology , silicon , competitive binding , chemistry , porosity , reuse , fabrication , materials science , organic chemistry , engineering , biochemistry , medicine , receptor , alternative medicine , pathology , waste management
The inexpensive fabrication and intrinsic optical and filtering properties of porous silicon (PSi) make optical PSi biosensors ideally suited for diagnostic testing. Long term goals of a diagnostic sensor would benefit from reversible binding chemistry to allow retesting of patient samples and collection of captured target for analysis. This work illustrates a versatile method to immobilize hexahistidine (His‐6) tagged molecular probes and utilizes reversible competitive chelating chemistry to reuse the porous silicon sensor device. Reuse of this Ni–His‐6 chemistry in a PSi affinity‐based biosensor was demonstrated with two different His‐6 tagged proteins to give consistent probe loading values within mimimal error over 4 cycles of use. Advantages of utilizing any site‐directed immobilization method include the homogeneity of surface bound probe molecule orientation, which we also illustrate resulted in a 2‐fold increase in detection capability of a secondary target antibody. As the current sensitivity range of macroporous PSi sensors (diameters of 30–100 nm) pose limitations to their use in medical applications; this work identifies the ability to improve target capture through the development of an efficient and versatile linking chemistry. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)