Bioluminescence

Bioluminescent Reporter Proteins: Three Classes, One Powerful Toolkit

Bioluminescent organisms produce three classes of proteins with properties that make them ideal reporter genes. Researchers use them to study cellular processes, map metabolic pathways, and run high-throughput screening (HTS).

Luciferases

Luciferases catalyze the enzymatic oxidation of a luciferin substrate — most notably Coelenterazine — producing light as a byproduct. Unlike first-generation firefly luciferase systems, our Coelenterazine-based luciferases (NanoLight™) don’t require accessory high-energy molecules like ATP or Coenzyme A. That difference greatly simplifies their use across reporter applications.

Gaussia Luciferase: Small, Bright, and Hard-Won

One of the smallest and brightest Coelenterazine luciferase we offer is Gaussia princeps luciferase. Discovering it took many research trips to sea, hunting for tiny 5 mm copepods that live sparsely scattered at depths of 700–900 meters. Collecting enough of them in pristine condition to build functional expression-cloning libraries was extremely laborious and expensive.

Among other applications, Gaussia luciferase enables researchers to observe single neuronal vesicle release, making it possible to image depolarization and track the release of insulin and other fusion proteins directly and quantitatively.

Photoproteins 

Photoproteins (Aequorin, Obelin) come “pre-charged” with Coelenterazine and molecular oxygen. When calcium triggers oxidation, they emit a flash of light. They typically need a reducing environment and very low calcium levels — otherwise they behave as poor luciferases in calcium’s presence.

Green Fluorescent Proteins (NanoFluor™)

Green Fluorescent Proteins fluoresce intrinsically, thanks to a unique peptide-derived chromophore that self-assembles after translation. GFPs shift the wavelength of luciferase or photoprotein signals from blue to green, enabling whole-cell studies of gene expression and visualization of drug-target interactions over time and across dosages.

BRET: A New Class of Biosensors

Our unique GFP and luciferase properties earned our scientists a Phase I SBIR grant to evaluate BRET (bioluminescent resonance energy transfer) as a novel biosensor technology. BRET has broad potential for HTS, medical diagnostics, and basic research. Its key advantage: it reveals both spatial and orientation information when a binding or proximity event occurs.

Proven in the Field

One sea pen GFP, Ptilosarcus, has the highest natural quantum efficiency of any fluorescent protein known. We’ve codon-optimized both Ptilosarcus and Renilla GFP, and they’re now in use for High Content Screening by Cellomics, as well as by Molecular Devices and Rigel Pharmaceuticals.

Untapped Potential

NanoLight Technology believes basic researchers, drug discoverers, and the agrichemical industry are only beginning to harness the power of naturally occurring bioluminescence in their research, screening, and discovery work.

Most of the visible spectrum measurable by standard luminometers remains unexplored — and NanoLight™ is well-positioned to develop bioluminescent reporter systems for it. These systems integrate easily into ELISA detection, direct in vivo imaging, cell tracking, DNA-DNA, DNA-RNA, and nucleotide-peptide detection, using a variety of strategies including split luciferase techniques. Because our intellectual property is solid, life science companies can trust that their discoveries — and any patents built on NanoLight technology — rest on secure ground.