About Sharp Edge Labs

The Fluorogen Activation Modules (the "FAMs")

Sharp Edge is bringing a new class of targeted biosensors to drug discovery and trafficking biology. Fluorogenic Activating Modules (or FAMs)* provide a more accurate and detailed detection method for cellular trafficking that is amendable to high-throughput screening and more detailed follow-up.

How the FAMs work

The FAMs are single-chain antibody fragments (scFvs) of approximately 140 amino acids that have been selected to bind and activate the fluorescence of specially designed dyes (the Flurogens).(1) FAM-fusions are created that behave similarly to GFP-tagged proteins. However, there are several advantages to the FAM technology. The ability to genetically target the FAM by creating fusion proteins (Targeting), combined with the ability to modulate the properties of the Flurogens (Sensing)) leads to a variety of new types of assays. For example, Sharp Edge Labs has developed a new set of cell-surface protein trafficking assays that are more detailed, and yet simpler than competing technologies.

Trafficking: Targeting + Sensing

By targeting the FAM to a cell surface protein (a GPCR for eaxample), and using cell impermenant and cell permeant dyes, we've created an easy and direct assay for receptor internalization.


Note that se-Red-xc is excluded from the cytoplasm, and therefor provides nicely selective labelling for the plasma membrane component of the target protein. Note also that unlike a surface ELISA, no washing is necessary because the Fluorogen is dark until it binds the FAM. The figure below shows this comparison in a live cell assay.

se-Red-s labels both surface protein and protein in the biosynthetic pathway (the ER/Golgi in this figure) and measures total protien, similar to GFP. The ability to switch dyes (and even add se-Red-S after imaging with se-Red-xc) provides and easy way to untangle cell-surface from total protein, which is difficult with other labelling methods.


A variety of fluorogenic sensor dyes has also been created, allowing targeted sensing, which creates a powerful tool for understanding trafficking. By using an extracellular pH Sensing Fluorogen, you can get a clean and uniqe signal from surface protein that has entered the endocytic pathway.

In this case, the pH sensor dye changes "color" when it moves from the neutral environment of the extracellular space, to the acidic pH of the endosome. Note that the sensor dye only reads out the pH at the location of the FAP, since it is dark otherwise (that is, it is a true, targeted sensor). This assay also requires no washing, and is amenable to live-cell and fixed cell preparations.

These features are especially useful for studying trafficking of well-known drug targets such as GPCRs, Ion Channels, and Transporters.

*The FAMs are covered by US Patent 8,664,364 and other US and international pending patent applications.

(1) Szent-Gyorgyi C, Schmidt BF, Creeger Y, Fisher GW, Zakel KL, Adler S, Fitzpatrick JA, Woolford CA, Yan Q, Vasilev KV, Berget PB, Bruchez MP, Jarvik JW, Waggoner A. Fluorogen-activating single-chain antibodies for imaging cell surface proteins. Nature biotechnology. 2008;26(2):235-40. Epub 2007/12/25. doi: 10.1038/nbt1368. PubMed PMID: 18157118.

(2) Fitzpatrick JA, Yan Q, Sieber JJ, Dyba M, Schwarz U, Szent-Gyorgyi C, Woolford CA, Berget PB, Waggoner AS, Bruchez MP. STED nanoscopy in living cells using Fluorogen Activating Proteins. Bioconjugate chemistry. 2009;20(10):1843-7. Epub 2009/01/01. doi: 10.1021/bc900249e. PubMed PMID: 20976031; PubMed Central PMCID: PMC2957894

(3) Szent-Gyorgyi C, Schmidt BF, Fitzpatrick JA, Bruchez MP. Fluorogenic Dendrons with Multiple Donor Chromophores as Bright Genetically Targeted and Activated Probes. Journal of the American Chemical Society. 2010;132:6.

The Sharp Edge Team
Scott F. Sneddon, Ph.D., J.D. Company President & CEO Scott holds a Ph.D. in Chemistry & Biophysics from Carnegie-Mellon University, a J.D. from Columbia University Law School and has over 20 years experience in the drug discovery industry, having held leadership positions at Pfizer and Genzyme. At Pfizer Dr. Sneddon was a member of the New Leads Discovery group under Fred Vinick. He then went to Genzyme with Fred to help establish Genzyme's small molecule drug discovery program. There he led the Assay Development and High Throughput Screening group and was a pioneer in implementing high-throughput functional cellular assays for primary drug screening (before such a thing was fashionable). He has worked as an attorney handling venture financing, licensing and ongoing strategic operation for startup and growth-phase companies in the biotechnology sector. He is also a registered patent attorney licensed to practice before the US Patent and Trademark Office.

Marcel Bruchez, Ph.D. Scientific Co-Founder & Chief Technology Officer Marcel P. Bruchez is a recognized leader in developing and commercializing research tools for bionanotechnology - an emerging field that creates or adapts materials and chemical processes to solve biological problems. As a graduate student, he modified quantum dots - nanometer-sized crystal particles - so that they could be used to tag proteins and label cells. Based on this work, he founded Quantum Dot Corporation to develop and commercialize quantum dots for biological applications. In 2005, Quantum Dot was purchased by Invitrogen Corporation, and Bruchez joined the Molecular Biosensor and Imaging Center as Program Manager for the National Technology Center for Networks and Pathways at Carnegie Mellon University. Research in his lab is focused on optical tools for detection of complex biological processes. They integrate nanotechnology, organic chemistry, and molecular biology tools to develop a wide range of probes for particular environments. There are three major programs that are currently underway: Quantum dots for real-time imaging in tissue regeneration and tumor biology. Dye-based structures for signal enhancement in sensors and fluorescent molecules.Expressible probes for in-vivo and live-cell imaging and sensing. These tools are all designed to investigate biological changes, in real-time, as they occur in living cells and animals.

Alan Waggoner, Ph.D. Scientific Co-Founder & Chief Architect Dr. Waggoner's research has focused on development of fluorescence-based detection systems for biology and biotechnology. The cyanine dye fluorescent labeling reagents developed in the laboratory have become widely used in industry and academic research for multicolor analysis of proteins, nucleic acids, cells and tissues with imaging microscopes and flow cytometers. Dr. Waggoner is currently leading the Molecular Biosensor and Imaging Center into development of microbiosensors for studying protein regulatory processes in living cells and tissues. The Center also has a NASA project for detecting sparse microorganisms in extreme environments.