Octet RED-Related Presentations at SBS Label-Free
Technologies Symposium
June 10-11, 2008 Dresden, Germany
Rapid and Versatile Characterization of Small Molecule Kinetics using Biolayer Interferometry
Eric Martin, Novartis, Emeryville, CA, USA
Biolayer Interferometry (BLI) provides label-free, real-time measurements from proteins to small molecules for kinetics characterization and screening. Affinities can be measured ranging from pM to mM with association and dissociation rates for better characterization and selection of drug candidates. Good agreement was found was found with SPR, ITC, NMR, and high concentration biochemical screening methods for small molecules down to MW=150. Applications included fragment-based screening, screening of inactive enzymes, and protein quantitation in cell lysates.
Applying Label-Free Technologies to Validate Screening Hits
David F. Mark, Ph.D., Hoffmann-La Roche Inc., Nutley, NJ, USA
Label-free direct binding technology is the method of choice for confirming that screening hits bind directly to the target of interest. A number of such technologies are available, such as, NMR, SPR, Thermo-shift and Biolayer Interferometry. A hit validation workflow will be described that takes advantage of the unique properties of each label-free technology to confirm the binding of hits to the target proteins. In particular, we will discuss how we have incorporated the ForteBio Octet RED System, a fiber optic based biosensor, into our hit validation workflow.
Hit Validation Using Biolayer Interferometry
Frank Podlaski, Hoffmann-La Roche Inc., Nutley, NJ, USA
ForteBio’s Octet RED System utilizes Biolayer Interferometry, a fiber optic based biosensor technology that can perform kinetic analysis of small molecule binding in an 8-channel format that is cost effective and easy to use. Its non-fluidic, simple, drop and read configuration give it advantages for quick assay development and may be better suited for analysis of complex mixtures or compounds with low solubility. This tutorial will describe our work evaluating the Octet RED System for hit validation while highlighting pros and cons of its current configuration.
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New Publications in Press
Detecting DNA: Getting and Begetting Cancer
AM Farkas et al., Current Opinion in Investigational Drugs, 8:981-986, 2007.
Determining Kinetics and Affinities of Protein Interactions using a Parallel Real-time Label-Free Biosensor, the Octet
Y Abdiche et al., Analytical Biochemistry, 377:209-217, 2008.
A Rapid Method for Determining Dynamic Binding Capacity of Resins for the Purification of Proteins
L Lerner et al., Protein Expression and Purification, 60:147-150, 2008.
Probing the Binding Mechanism and Affinity of Tanezumab, a Recombinant Humanized anti-NGF Monoconal Antibody, Using a Repertoire of Biosensors
Y Abdiche et al., Protein Science, Punished online doi:10.1110/ps.035402.108. |
New Technical Notes for use with the Octet QK and Octet RED Systems
ForteBio are pleased to offer four new Technical Notes for download from the support library on www.fortebio.com. Octet Serial number registration is required to download technical notes.
TN-3010: Batch Immobilization of a Biotinylated Ligand onto Streptavidin Biosensors
This technical note outlines a general batch mode procedure for immobilizing a biotinylated ligand onto Streptavidin biosensors in the biosensor tray assembly on the benchtop. The goal is to develop a procedure that creates a binding surface that yields maximum and reproducible response. The protocol outlines how to determine the optimal concentration and time for immobilization online, and then describes how to transfer these parameters to offline immobilization.
TN-3011: Biotinylating Antibody in Stocks Containing Carrier Protein
This technical note provides a detailed protocol for isolating antibodies from carrier protein and subsequently biotinylating the purified antibodies.
TN-3012: Biotinylating Very Small Quantities of Protein for Immobilization onto Streptavidin Biosensors
This technical note provides guidelines for biotinylating a protein of interest for use with Streptavidin biosensors on the Octet QK or Octet RED System, when <150 mg total protein is available.
TN-3013: Development of a Quantitation Assay using His-Tag Biosensors
Label-free quantitation assays for His6-tagged proteins can be developed using the Octet QK or Octet RED instruments with Streptavidin biosensors. The flexible design of the biosensor tray assembly enables batch immobilization of nickel-activated Biotin-NTA or an anti-His directed antibody onto Streptavidin biosensors outside the instrument. The ligand-coated biosensors can be used to rapidly quantitate protein(s) that are expressed with the His6-tag. (available August 2008) |
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