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News

October 22, 2019

We are happy to announce that the University has acquired a Applied Photophysics Chriascan V-100 Circular Dichroism Instrument and a Nanotemper Dianthus NT2.3 pico Instrument.

The Chirascan instrument is especially good at detecting changes in secondary and tertiary structure induced by any change in the solvent environment or ligand binding. We are the only University in the greater metropolitan area to have such an instrument for general use. It is frequently used  to perform quality control studies on proteins and peptides. Our instrument is equipped with a heat ramp, so that one can observe dynamic changes in the spectra during unfolding. This allows for the measurement of ligand binding, and enables stability studies among other things.

We foresee the Dianthus instrument to be be useful for many types of experiments, including microplate based testing for  small-molecule/target and antibody/epitope interactions. We are the first University to have such an instrument.  It can measure the dissociation rate constants of most interactions of pure and partially pure biomolecules. The technology requires a fluorescent-labelled receptor or epitope to act as a molecular beacon for changes in the fluorescent properties based on molecular environment induced by a rapid, mild heat pulse. Because of the sensitivity of fluorescence, this instrument can achieve a throughput of about 760 samples per hour and with reduced protein use compared to other biophysical techniques.  The purchase of a specialized microplate is required.

The product descriptions can be found at

https://www.photophysics.com/systems/chirascan-systems/chirascan-v100/system-information/

September 12, 2019

We are happy to announce that the University has acquired a new high-throughput multi-function microplate reader for all to use. It is called the “Biotek Synergy NEO2- TRF” and is capable of performing AlphaLisa, Luminescence,and various forms of Fluorescence assays in a 96, or 384-well format.

https://www.biotek.com/products/detection-multi-mode-microplate-readers/synergy-neo2-hybrid-multi-mode-reader/

It is especially sensitive for time-gated FRET experiments, because it is equipped with laser excitation tuned for the most common fluorophores. It is also equipped with a 50-plate automated loading system. It is located in Bronk room 214 and is now bookable on the PPMS system (https://ppms.us/rockefeller/login/?pf=4) for $19.16 per hour charged to your grant. If you have any questions, or would like to be trained on this instrument, please feel free to contact us.

October 18, 2018

The HTSRC is now offering a fragment-based screening platform based on 1056 fragments purchased from LifeChemicals, with measured high solubility in aqueous media. Each fragment represents a cluster, from which analogs of hits can be purchased for initial S.A.R. studies. Our basic platform includes 240uM- 1mM primary screening using DSF, followed by hit confirmation using either MST, SPR or ITC. Functional assay readouts are also amenable to this form of compound screening.

We have also made significant additions of 705 compounds to our drug re-purposing libraries and annotated pharmacologically active compounds.

March 20, 2018

The Tri-Institutional Therapeutic Discovery Institute has donated a 100,000 member compound collection from ChemDiv. This is a maximally diverse set and a great point for a rapid start to generate biological data. It nicely covers chemical space from a larger collection of ChemDiv molecules (collection of 1.6M compounds) and is based on a few thousand unique chemotypes in ChemDiv’s collection. This 100,000 compound set includes representative samples from ChemDiv’s Discovery Chemistry (DC), New Chemistry (NC) and Innovative Chemistry (IC) collections.

August 23, 2016

A new SeaHorse XF96 has been installed. This instrument is useful in studying cell metabolism, as it can kinetically measure the dissolved oxygen and pH of the cell media surrounding cells cultured in 96-well plates.  pH changes and oxygen consumption correlate with anaerobic and aerobic metabolism. The system can be used for characterizing the effects of compounds or genetic modifications  in cell lines  and for examining potential toxicities of hit compounds using extracellular acidification rate and oxygen consumption rate analysis. It is additionally useful for studying adipose function and differentiation and other aspects of mitochondrial and glycolytic function in living cells and isolated mitochondria.

We recently installed a new Differential Scanning Fluorimeter (DSF) system (http://www.ncbi.nlm.nih.gov/pubmed/17853878).  This instrument can be used to measure the binding affinity of small molecules to proteins. The instrument we have is called the Bio-Rad CFX 96 (http://www.bio-rad.com/en-us/product/cfx96-touch-real-time-pcr-detection-system) generates a thermal ramp in the presence of an environmentally sensitive fluorescent dye on 96-well plates and measures the changes fluorescence intensity of the sample. A shift in the melting curve happens when the protein is bound to a ligand. The melting temperature dependence on ligand concentration can be fit to derive an affinity constant or Kd. It can also be used to perform qPCR in a 96 well format.

6000 new screening compounds have been purchased, from Edelris, ChemBridge and ChemDiv. These compounds were chosen to differ from the existing HTS screening collection and to represent diverse high quality scaffolds based on QED scores and fsp3 scores.  The Edelris compounds have been designed to mimic themes from naturally derived compounds and analogs of any hits can be obtained from Edelris.

The annotation of the HTSRC library now includes REOS filters, QED and fsp3 scores so that screening hits can be ranked according to these, among many other standard medicinal chemistry metrics already available in CDD.

March 10, 2015

We recently acquired a Wyatt Dynapro plate reader which is a high-throughput dynamic light scattering instrument. The system can read 384-well plates and is ideally suited for measuring aggregation and solubility (average particle size) of small molecules and biomolecules. We routinely use this to test all of our screening hits as a quality control measure to ensure that the compounds are soluble at the concentration needed for the assay, and to ensure that there are no compound mediated protein aggregation artifacts. The instrument also can be used for polymerization dynamics.

We now have a refurbished Agilent HPLC system, coupled to a Photo-diode array detector and an Agilent time-of-flight mass spectrometer. We routinely use this to ensure compound purity and integrity, but it can be useful to those interested in analyzing compound samples for purity and composition.

We have added about 800 Selleck and 11,000 Enamine compounds to our compound collection. The Selleck compounds augment our current collection of 3000 “annotated” compounds, meaning that these are compounds with known references in the scientific literature. The Enamine compounds represent expanded diversity, but also have very high medicinal chemistry scores and are available at low-cost for re-supply and commercially available analogs for structure activity relationship determination.

We have decommissioned the Union Biometrica COPAS, particle sorting instrument, which we found to be a rarely-used and difficult to maintain, suffering from many breakdowns.

May 2, 2014

The new Rapid-Fire (Agilent) mass spec-based screening system is now operational. This system allows one to perform rapid assays using accurate mass spectrometry of solid-phase extracts. We are very excited about this technology, as it enables label free detection of enzyme producs and substrates and cell metabolites. Unofortunately, we are fully booked on this platform for the time-being.

The TECAN EVO150 is the latest liquid handling instrument installed at the HTS Resource Center. This multi-functional liquid handler is equipped with an MCA384 pipetting head to perform microtiter plate-to-plate transfers in both 96- and 384-well plate formats from as low as 1uL to 200uL. The EVO150 is also built with an 8-channel liquid handling arm called the LiHa. This liquid handling arm offers parallel pipetting from 1uL up to 1000uL. The instrument has high-throughput plate stackers that can handle up to 100 plates, a barcode reader, and a disposable tip washer. With this setup, one can quickly replicate plates, cherry pick and re-array samples, perform automated serial dilutions, and with the center’s assistance, build custom applications. The TECAN EVO150 has replaced the Janus Mini.

As with any of our services, please use the online suggestion box to let us know how we can improve.

October 19th, 2012

We are happy to announce several improvements to the capabilities of the High-throughput Screening Resource Center, which are available to you, and present you with a summary below. Please feel free to contact our staff if you have any specific questions.

Improvements in capability for the analysis of biomolecular interactions

We have installed a Proteon XPR surface plasmon instrument. This instrument is more sensitive (mw> 100 versus 300) than our current Biacore 3000 and has higher throughput, allowing for 36 interactions with controls, simultaneously, versus 4 in the biacore. We are happy to train anyone interested and help in the design of experiments.

A Nanotemper microscale thermophoresis instrument has been installed. This instrument can measure biomolecular interactions (equilibrium binding) in a very small volume (8 ul), provided one of the interacting molecules is fluorescently labeled. As contrasted with fluorescence polarization or Surface Plasmon Resonance, there are no specific limits of molecular weight; however, the binding event must effect the interactions of the biomolecule with the solvent system, through changing mass, charge and/or solvation shell, such that its migration through a temperature gradient is affected. We are happy to train anyone interested in this simple technique and help in the design of experiments.

Improvement in Cellular Assays and Western Blots

A LICOR Odyssey SA is an infrared laser scanning instrument that allows for a quantitative determination of antibody binding to western blots or to fixed cells in a microplate. The instrument is capable of reading 50, 384 well microplates in 90 minutes and can scan two colors simultaneously, thus allowing the introduction of normalization controls in every sample. The system has a much larger linear range than chemiluminescent systems typically used in such applications.  This is a very useful instrument for measuring antigen expression in both cells and in purified samples.

 

 



Contact

The Rockefeller University
Box 203
1230 York Ave
New York, NY 10065

High-Throughput Screening Resource Center
Bronk Laboratory
DWB Room 216