Reusable linking chemistry for His-6 tagged proteins in an affinity-based porous silicon biosensor
✍ Scribed by Bonanno, Lisa ;DeLouise, Lisa
- Book ID
- 105365245
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 352 KB
- Volume
- 206
- Category
- Article
- ISSN
- 0031-8965
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
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)