Genetic engineering of a single-chain antibody fragment for surface immobilization in an optical biosensor.

Journal Article (Journal Article)

The development of a biosensor based on a genetically engineered biomolecule offers many potential advantages to sensors that rely on natural proteins only. Here we present how protein engineering techniques can be used to introduce a functional unit for surface immobilization into a single-chain antibody fragment (scFv). A peptide known to mimic the binding properties of biotin was fused to the carboxyterminus of the phosphorylcholine-binding scFv fragment of IgA McPC603. This fusion protein could be immobilized on a streptavidin monolayer. The resulting scFv monolayer was capable of binding a fluorescently labeled phosphorylcholine analog, as detected by total internal reflection fluorescence. In contrast, an scFv monolayer formed by introducing biotin through chemical modification was not capable of binding phosphorylcholine. These results demonstrate the utility of site-specific, oriented attachment strategies in the formation of protein monolayers in optical sensors, made possible by the use of protein engineering techniques.

Full Text

Duke Authors

Cited Authors

  • Piervincenzi, RT; Reichert, WM; Hellinga, HW

Published Date

  • March 1, 1998

Published In

Volume / Issue

  • 13 / 3-4

Start / End Page

  • 305 - 312

PubMed ID

  • 9642767

International Standard Serial Number (ISSN)

  • 0956-5663

Digital Object Identifier (DOI)

  • 10.1016/s0956-5663(97)00130-9


  • eng

Conference Location

  • England