Genetically engineered luminescent proteins in biosensing

Laura Rowe, Mark Ensor, Daniel Scott, Sapna Deo, Sylvia Daunert

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Scopus citations

Abstract

Luminescent proteins originally isolated from marine or terrestrial organisms have played a key role in the development of several biosensing systems. These proteins have been used in a variety of applications including, immunoassays, binding assays, cell-based sensing, high throughput screening, optical imaging, etc. Among the luminescent proteins isolated, the bioluminescent protein aequorin has been one of the proteins at the forefront in terms of its use in a vast number of biosensing systems. In our laboratory, we have employed aequorin as a label in the development of highly sensitive assays through chemical and genetic modifications from single step analysis of physiologically important molecules in biological fluids. An important aspect of optimizing these assays for clinical use involves understanding the stability of the various aequorin variants that are available. To this end we have designed several stability studies involving three important aequorin mutants, Mutant S, Mutant 5, and Mutant 53. The cysteine free aequorin, Mutant S, has been the most ubiquitously used aequorin variant in our laboratory because of its increased stability and activity as compared to native aequorin. Mutant 5 and Mutant 53 contain a single cyteine residue at position 5 and 53 in the protein, respectively. Because of the presence of a single cysteine residue, Mutant 5 and Mutant 53 both can be site-specifically conjugated. This site specific conjugation capability gives Mutant 5 and Mutant 53 an advantage over native aequorin when developing assays. Additional studies optimizing the expression, purification, and charging of aequorin Mutant S were also performed. A thorough understanding of the efficient expression, purification, and storage of these aequorin mutants will allow for the more practical utilization of these mutants in the development of future biosensing systems.

Original languageEnglish (US)
Title of host publicationGenetically Engineered Probes for Biomedical Applications
DOIs
StatePublished - Jun 26 2006
Externally publishedYes
EventGenetically Engineered Probes for Biomedical Applications - San Jose, CA, United States
Duration: Jan 24 2006Jan 24 2006

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6098
ISSN (Print)1605-7422

Other

OtherGenetically Engineered Probes for Biomedical Applications
CountryUnited States
CitySan Jose, CA
Period1/24/061/24/06

Keywords

  • Aequorin
  • Aequorin expression
  • Bioluminescence
  • Biosensors
  • Luminescent proteins
  • Photoprotein stability
  • Site-directed mutagenesis

ASJC Scopus subject areas

  • Engineering(all)

Fingerprint Dive into the research topics of 'Genetically engineered luminescent proteins in biosensing'. Together they form a unique fingerprint.

Cite this