TI-3D Facilities : Automation & High Throughput Screening Facility

The Texas Institute for Drug and Diagnostics Development is currently bringing on-line an High Throughput Automation Facility. This facility has been established to foster drug discovery and development through the fully automated compound synthesis and (screening of commercial chemical libraries as well as in-house compounds. As a part of the Automation facility, the High Throughput Screening Facility (HTSF) is designed to be highly flexible in order to meet the needs of multiple investigators performing a variety of assays. In addition we aim to provide cost-effective and time-efficient access to high throughput screening technology across the iUT-Campus as well as off-campus users. The facility staff assists assay development and miniaturization, willl train users to use some of the instruments such as a microplate reader and liquid handling robot, and provide consultation for data analysis.

ACKNOWLEDGMENT:
Users of the High throughput screening facility are urged to acknowledge any work performed by this Facility in their publications. Our service is a scientific support, not just a technical. Please send a copy of your publications to facility director, Eun Jeong Cho. We will compile the publications and may use as supporting material in the future for grant application such as Shared Instrumentation Grant application.
Examples:
1. Screening: “We thank The Texas Institute for Drug and Diagnostic Development (TI3D)-Automation and High Throughput Facility for their support on high-throughput screening and chemical libraries.”
2. Instrument use: “We thank The Texas Institute for Drug and Diagnostic Development (TI3D)-Automation High Throughput Facility for the access to the (instrument/s)”.

Instrumentation

Instrument Reservation

TI-3D users can reserve instruments from TI3D Scheduler. To reserve an instrument, you can log in and select the day and time you wish to use an instrument.

Screening Equipment

Perkin Elmer EnVision Microplate Reader (Website)

Application
General assay development utilizing various detection technology such as absorbance (UV-Vis), fluorescence intensity (FI), fluorescence polarization (FP), time-resolved fluorescence (TRF), luminescence (Lum), and Alphascreen.

Specs

  • User-changeable label-specific optical mirror modules and filters
  • Superior detection sensitivity and high measurement speed (< 1 min for 1536-well plate with one flash/well)
  • 1-1536 well microplate reading capability from bottom or top
  • Endpoint, kinetic, spectrum and well-area scanning read types
  • Temperature controlled (4-45°C)
  • Manual or automated serial plate reading via integrated plate stacker holding 50 plates
  • Filters

ManualUser ManualReference Manual

Perkin Elmer MicroBeta TriLux Microplate Scintillation and Luminescence Counter*(Website)

Application
Specific for radioactive assay utilizing liquid scintillation or luminescence detection.

Specs

  • Samples in microplates, tubes or on filters
  • Sample capacitites of up to 16 or 32 plates
  • Updated model is fitted with an external loading platform for fully automatic loading by robot systems

* Located in BME 6.328B

Jasco J-815 CD Spectrometer (Website)

Application
Specific for assay development for optically active biological molecules (type of sugar or amino acid) and secondary structure of macromolecules to analyze for activity.

Specs

  • Circular dichroism (CD) and UV-Vis absorbance (standard)
  • Measurement ranges from 163-900 nm, 1100 nm optional
  • 6-cell Petier thermoelectric temperature control for simultaneous Tm measurement of upto 6 samples (6-90°C)
  • Fluorescence accessory to measure fluorescence simultaneously
  • High throughput, temperature regulated automated sampling of up to 192 samples including thermal ramping and sample recovery capabilities

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Liquid Handling Equipment

Perkin Elmer Janus Workstation (Website)

Application
Laboratory automation workstation that performs versatile liquid handling tasks. Main task will be plate reformatting, large scale assay plating, and cherry picking.

Specs

  • Pipette tools consisted with 8-tip varispan and 96/384 multi-channel can transfer from 0.5 µL to 1 mL with high precision
  • Integrated stacker carousel can provide external supply and storage of microplates (50 plates), deepwell plates, and P20 and P250 pipette tips
  • Faster throughput achieved with multichannel tools
  • Flexible with easy adaptation of any labwares or tips

Art Robbins Instruments Phoenix (Website)

Application
Laboratory automation workstation specific for low volume (< 1 µL) liquid handling tasks. Main task will be library transferring and assay plate formation.

Specs

  • Simultaneous dispensing equipped with fixed 384 syringe head module and a single channel dispenser.
  • Fast liquid transferring speed (< 1 min) with a single channel head to 96 well plates.
  • Transfer liquids from 384-microwell source plates to 384/1536-microwell destination plates with very high accuracy from 100 nL to 100 µL.
  • 9 deck positions are available for plates and washing stations.

BioTek Microflo Select (Website)

Application
Self contained, user programmable dispenser machine that dispenses a wide variety of liquids to microwell plates.

Specs

  • Contains a peristaltic pump with eight individual tubes which transfer liquids from a supply bottle to various microwell plates.
  • Able to dispense volumes from 1 to 9999 µl using one of 3 cassettes.
  • Cassette sizes are 1 µl, 5 µl, and 10 µl.
  • Plate stackers are able to hold up to 30 plates each (input and output).
  • Standard carrier is able to accommodate 6-, 12-, 48-, 96-, 384-, and 1536-well plates that comply with SBS microplate standards.
  • Able to dispense a variety of solutions including buffered saline and reagent solutions.
  • Can store up to 99 dispense protocols.

Synthesis Equipment

CEM Explorer 48 Automated Microrave Synthesizer (Website)

Description
Many reactions which require forcing conditions (high temperatures and reaction times) can benefit greatly from microwave irradiation. The Explorer 48 is easily programmed to automatically run up to 48 reactions in sequence, using 4- 10- or 35- mL reaction vials.

CEM Liberty Automated Microwave Peptide Synthesizer (Website)

Description
The Liberty is an automated solid-phase peptide synthesizer which uses microwave irradiation to minimize reaction times and often give improved yields and purity over standard conditions. The system is capable of making up to 12 peptides a day at scales up to 5 mmol.

Mettler-Toledo MiniBlock Peptide Synthesizer (Website)

Description
The MiniBlocks system is used for convenient parallel synthesis with up to 48 reactions at once in small test tubes (smallest size 4.5 mL). The reactions can be sealed under an inert atmosphere and heated or cooled under a large range of temperatures, either on a standard stir plate (with MiniBlock XT) or on a rotary shaker. Solid-phase synthesis is especially convenient with the MiniBlocks, as reaction solutions can be drained and filtered out the bottom of each reaction concurrently. Parallel work-up (purification) with e.g. solid-phase extraction (SPE) cartridges is also supported. The MiniBlock platform is great for the synthesis of chemical libraries, but it is also useful for reaction optimization, especially when used with the MiniMapper liquid handling robot.

Mettler-Toledo MiniMapper liquid handler (Website)

Description
This liquid-handling robot can be easily programmed to add numerous solvents and reagent solutions to reaction vessels or microtiter plates. It can handle small quantities of corrosive reagents under an inert atmosphere. It is optimized for convenient use with the MiniBlock platform.

Combiflash Rf-200 UV-Vis Automated Flash Chromatography System (Website)

Description
A flash chromatography system with a 200-780 nm UV/Vis-variable wavelength PDA detector. Operating pressure capability of 200 PSI for reversed phase flash chromatography. Radio Frequency Identification technology to identify the columns & collection racks, eliminate errors, streamlines programming, and provides additional safety features by automatically setting pressure limits & fill levels. Automated, self-cleaning injection valve. Level sensing for inlets and waste container, vapor detection, and secondary containment of solvents and waste overflow prevention

Dionex Ultimate 3000 HPLC

Description
Analytical HPLC system, a 3 µm C18 column (4.6 x 150 mm) for reverse phase applications. UV-vis detection by a PDA (190-800nm). Temperature-controlled column compartment (rt-45C) and solvent degasser. Two mobile phase mixing. Sample introduction is accomplished through autosampler. Variable combination of sample containers are applicable (3 racks, each rack can hold 1.8, 2, or 10mL vial, 96 or 384well microwell), suitable pressure up to 7000psi, sample injection volume of up to 100uL

Agilent 1200 Series HPLC

Description
The Agilent LC system features a quaternary pump that provides mixing of up to four solvents and flow rates up to 10 mL/min. A 5 µm C18 column (4.6 x 150 mm) is connected for analytical, reverse-phase separations. The system is equipped with two detectors: a variable wavelength detector capable of monitoring a single wavelength between 190-600 nm and an evaporative light scattering detector (ELSD) that provides universal detection independent of an analyte’s absorbance profile. A temperature-controlled column compartment and solvent degasser are also available, and sample introduction is accomplished through the use of a manual injector and a 20 µL sample loop.

Shimadzu analytical and preparative HPLC systems

Description
Two custom designed Shimadzu HPLC systems for high throughput analytical and preparative work. Both systems have cooled autosamplers with adjustable configuration for either two 96-well plates, or up to 70 standard HPLC vials, automated eight-solvent and six-column switching and PDA detectors (190-800nm). Both systems are also enabled with dual rack 3.5 ml x 70 tube fraction collection racks. The preparative system pumps can deliver up to 20 ml/minute and the preparative collection system can also be modified for large volume fraction collection.

Genevac DD-4 centrifugal evaporator

Description
Conveniently removes both low- and high-boiling solvents (including DMSO) from test-tubes (e.g. generated by MiniBlock or prep. HPLC purifications) and microtiter plates.

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Thermo-Electron/Finnigan LTQ XL LC-MS (Website)*

Description
*Services are provided through Department of Chemistry, Mass Spectrometry Facility.

Other Equipment

Thermo Scientific ALPS 3000 Plate Sealer (Website)

Application
Securely seal various plate types, which contain either chemical libraries dissolved in 100% DMSO solution or assay reagents for high throughput screening.

Specs

  • Uses five (5) standard and two (2) custom seals including clear sealing for both polystyrene and polypropylene plates.
  • The range of sealing plate height is 8 -46mm.
  • Operating temperature range is 15-40°C
  • Cycle time is between 6 to 12 seconds (per plate) depending on plate types, sealing time can be adjustable from 0.5s to 5s.
  • Sealing temperature can be controlled within the range of ambient to 200°C.

Zebra Z4M Plus Bar Code Printer (Website)

Application
Preparing bar coded labels for all screening processes.

Specs

  • Print Methods: Thermal Transfer.
  • Auto-calibration.
  • Resolution: 203dpi.
  • Print Speed: 10″ (254 mm) per Second.
  • Print length up to 157″ (3,988 mm).

Eppendorf Centrifuge 5810R with Swing-bucket Rotor (Website)

Application
Sample re-hydration and spin down.

Specs

  • Variable Speeds to 14,000 rpm.
  • Variable Time from 1-99 minutes.
  • Temperature Control from -4 – 40° C.
  • Print Speed: 10″ (254 mm) per Second.
  • Capability: 18 x 1.5 and 2.0 ml Tubes, 16 Microplates, 2 Culture Plates, 4 Deepwell Plates, or 2 Deepwell Assemblies (filter and catch-plate stacks)

VWR Signature Incubating Microplate Shaker (Website)

Application
Can be used for defrosting and mixing samples from microwell plates.

Specs

  • Can shake up to four microplates or two microtube racks with adjustable temperature (5° C above ambient temperature to 65° C)
  • Speed range: 100 – 200 rpm
  • Timer: 1 second – 160 hours

TxSACT Program

Supporting scinetists with cancer-related screening and compound development granted by CPRIT in collaboration with GCC-CG

More Info

Chemical Library

Libraries are all formatted on 384-well, dissolved in 100% DMSO with ~10 mM concentration. Library plates are stored at -40° C Revco freezers.

Provider Name of Library Number of Compounds Number of Plates, 384 Well Plate Number
Chembridge Kinase Set 11,250 36 TI3D_0001 – 0013
TI3D_0027 – 0049
Fragment Set 4,000 13 TI3D_0014 – 0026
NIH NCC Clinical Collection 446 2 TI3D_0050 – 0051
MicroSource Discovery Systems, Inc. Spectrum Collection 2,000 7 TI3D_0097 – 0103
Maybridge HitFinder V. 9 11,400 45 TI3D_0052 – 0096
UT-Austin Martin Collection 800 2 TI3D_SFM01 – 02
Chemdiv Diversity Set 12,900 40 TI3D_0133 – 0173
Halopyridine 1,180 4 TI3D_0174 – 0177
Brominate 163 1 TI3D_0178
Non-Brominated Fragment Set 12,800 40 TI3D_0179 – 0218
Target Focused Kinase Set ~600 2 TI3D_0106 – 0107

Please refer to the wiki for more information.

Data Analysis

TI-3D currently uses Collaborative Drug Discovery (CDD) for their Data Analysis needs. CDD’s secure web-based database system allows researchers to archive, mine, and collaborate with both other researchers in the lab and internationally. The application’s many features include compatibility with Excel, making it easy to store and download data to a public format; and the ability to restrict data or make it public, allowing researchers to both secure their data and share it with others.

CDD Application Tutorial
Batch Upload File for Tutorial

Services and Fee Structure

  • NOTE: Fee schedule is now active, starting from April 1, 2009.
  • Fee Schedule is applied to both on-campus and off-campus users.
  • Off-campus users should contact the TI3D-HTSF Staff.
  • Pricing is highly dependent on the number of steps involved, the degree of multiplication, the length of preparation, and a number of other factors. Please contact TI3D-HTSF for project-specific pricing.
  • Services: Consultation, training, compound screening, cherry picking, and preliminary data analysis

Please refer to the wiki for more information.

Publications

  • Synthesis and evaluation of quinoxaline derivatives as potential influenza NS1A protein inhibitors
    You L, Cho EJ, Leavitt J, Ma LC, Montelione GT, Anslyn EV, Krug RM, Ellington A, Robertus JD. [pdf]
  • Selective Inactivation of a Human Neuronal Silencing Phosphatase by a Small Molecule Inhibitor
    Zhang M, Cho EJ, Burstein G, Siegel D, Zhang Y. [pdf]
  • A Continuous, Fluorescent, High-Throughput Assay for Human Dimethylarginine Dimethylaminohydrolase-1
    Linsky T, Fast W. [pdf]
  • Discovery of Halopyridines as Quiescent Affinity Labels: Inactivation of Dimethylarginine Dimethylaminohydrolase
    Johnson CM, Linsky TW, Yoon DW, Person MD, Fast W. [pdf]

Location & Contact

Mailing Address
The University of Texas at Austin
1 University Station
Austin, TX 78712
Mail Code: A5300
Lab Location
The University of Texas at Austin
Robert A. Welch Hall
Room 3.316
Lab Phone: (512) 471-5485
Map on Campus
Google Map
Contact
Dr. Eun Jeong Cho
1 University Station
Austin, TX 78712
Mail Code: A5300
Office Phone: (512) 471-5507
Office Location: WEL 3.310