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Mid Atlantic Chapter



April 2003
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The Laboratory Robotics Interest Group
Mid Atlantic Chapter

April 2003 Meeting

Emerging Technologies

Date:        April 9, 2003
Place:      Somerset Marriott Hotel, 110 Davidson Ave., Somerset, NJ 08873
                    Phone: 732-560-0500, Fax: 732-560-3669
Itinerary:  Social Period -  5:00 to 6:30 PM
                  Meeting & Presentations -  6:30 to 9:00 PM  
Registration: REQUESTED, not required.  Registering will allow us to more accurately gauge seating requirements and refreshment needs.  Register on the web at https://www.lab-robotics.org/member/meetings.asp?rid=1There will be drawings from the web registrants for our beautiful LRIG laser pointers and photon keyring lights and any other donated (hint, hint) prizes.

Door Prizes:
Laser Pointers (LRIG)
Photon Keyring Lights (LRIG)
Door prizes for the drawing gratefully accepted!
 

Agenda:  
Attend this program to hear short presentations on new laboratory automation tools and technologies. Based on a similar meeting held in Palm Springs, Emerging Technologies Showcase at LabAutomation2003, this meeting will focus on the scientific and productivity gains of these technologies rather than the business aspects.

Food and refreshments will be available FREE OF CHARGE during the Social Period. We thank IGEN International, Inc. for subsidizing the food and refreshments at this meeting.

There is always a Job posting board at the social. Please encourage your recruiters to give you material to post and distribute. Openings may also be posted at https://www.lab-robotics.org/forum/default.asp?CAT_ID=2.

There is no fee to attend the meeting.

Presentation:  Double-Sink?PAMPA: The High Throughput Gastrointestinal Absorption Model for 21st Century Drug Discovery
Cynthia Berger, Ph.D.; pION Inc.; http://www.pion-inc.com/

The need for high-throughput, low-cost permeability screens to augment the use human cell cultures has spurned the innovation of the Parallel Artificial Membrane Permeability Assay (PAMPA). The acceptance of PAMPA has increased in the pharmaceutical industry and several PAMPA models have been shown to correlate well with passive gastrointestinal tract (GIT) absorption.

Several models make use of multiple pH measurements to explain in vivo results found with acidic and basic compounds. To simplify the experimentation and calculations, both acceptor and donor compartments are normally the same pH (iso-pH). However, these measurements do not accurately reflect the true underlying process that drives permeability: pH and chemical sink.

In this work, we describe a new PAMPA model called the ouble Sink?to simulate pH and chemical sink and provide a closer correlation with actual human jejunal conditions. This new protocol uses a pH gradient between the acceptor and donor wells (the first sink) and a chemical scavenger system present in the acceptor wells (the second sink). This model is beneficial for high throughput screening because it operates on a faster time scale than traditional PAMPA and has greatly reduced membrane retention. The later fact facilitates quantitation of highly lipophilic compounds produced in today drug discovery work, a particular problem with iso-pH studies.

Presentation:  Upping the Ante in Low Volume Pipetting: Can we Really Handle Nanoliter Volumes in the Real Screening World?
Dean Colone, Yan Kong, Mary Jo Wildey, Ph.D., (Johnson and Johnson PRD) Gregory J. Wendel, Ph.D., Rick Bernal, George Grubner, (Zymark Corp.) Marc Boillat, Bart van der Schoot, Ph.D. (Seyonic Inc.)

Nanoliter dispensing will prove beneficial in efficiently managing compound libraries and reagents, lowering overall screening costs, and minimizing waste. Challenges are moving small volumes accurately and precisely, issues such as clogging, handling viscous liquids, minimizing dead volumes, washing tips and individual tip dispensing. Working with the Beta Zymark inl10 Sciclone system we are now capable of dispensing 25nl into microtiter plates with a total volume of 10uls with CV < 10%. Attached to the each dispenser is a microflowmeter controlled closed loop system that is capable of aspirating and dispensing 10nls to 10,000nls. Tips can either be fired simultaneously or independently and the system reports volumes actually transferred. This presentation will outline our experiences with the validation of these low nanoliter dispensing systems, using both dye validators as well as test screening assays. Concepts on how this type of dispenser unit may fit into an HTS/uHTS production facility will also be discussed.

Presentation:  RNA-based on-bead screening to identify small molecules that modulate gene expression
Matthew C. Pellegrini1, David Dunn2, Shah Ikramul Huq1, Andrew Cole2, Edward Heebner2
1PTC Therapeutics, Inc., 100 Corporate Court, South Plainfield, NJ 07080
2Pharmacopeia, Inc., PO Box 5350, Princeton, NJ 08543-5350

RNA is a single-stranded nucleic acid and, much like proteins, can fold into unique structures that are capable carrying out multiple functions. In addition to its key role in being the messenger of gene expression, there are now many examples of RNA structures that function as key regulators of gene expression, or as part of complexes that have enzyme activities. These properties make RNA an exceptional target for drug discovery. The challenge is to develop appropriate screening technologies and chemical libraries that lead to the identification of small molecules that interact directly with RNA and affect gene expression; PTC Therapeutics, Inc., has developed a Targeted Ribonucleic Acid Chemistry (TRAC) technology platform for just this purpose. One TRAC technology utilizes on-bead screening (OBS) of combinatorial chemical libraries to identify small molecules that bind directly to RNA structures. The bead-based library is rapidly screened against multiple RNA targets utilizing RNA that has been tagged with a fluorescent compound. In this procedure, fluorescent RNA-bound beads carrying active compounds that bind the target RNA are readily separated from those carrying inactive (non-binding) compounds, and their unique chemical structures are determined by biophysical techniques.

Pharmacopeia, Inc., has developed a large and diverse bead-based library of drug-like compounds using ECLiPSTM technology (ncoded Combinatorial Library on Polymeric Supports?. The work presented herein describes the use of TRAC technology to screen a portion of the Pharmacopeia collection for compounds that bind to medically relevant RNA targets. ECLiPSTM beads were allowed to react with fluorescent-labeled target RNA and the chemical structure of the active compounds was determined by gas-chromatographic analysis of the chemically encoded tag, attached to the beads during the combinatorial synthesis. Active compounds were resynthesized and characterized in vitro using a functional assay (Transcription-linked-Translation). The re-synthesized compounds maintained their activity when tested in solution and showed high specific binding to one of two target RNAs. Furthermore, the compounds specifically inhibited RNA-dependent gene expression in a cell-free in vitro translation assay. Thus, the combination of ECLiPSTM and TRAC technologies provides a robust, drug discovery platform technology that enables the discovery novel chemical scaffolds that interact specifically and selectively with a myriad of RNA-based drug targets, and automation of the screening assay can be accomplished using the fluorescent-labeled RNA and flow cytometry (FACS).

Presentation:  
Roeland Papen, Picoliter Inc., http://www.picoliterinc.com/

Picoliter Inc. is a development stage company with a novel technology for precisely moving and sensing very small volumes of liquids including compounds dissolved in DMSO, proteins and live cells. We are using this technology to enable a broad range of tools for life science researchers. Our current path is to focus on providing nanoliter dispensing instruments for HTS applications, as well as enabling emerging applications in cell- and tissue-based research.

Presentation:  Rb+ efflux assays with the ICR12000 to study K ion channels
Tina Garyantes, Junjie Xiong; Aventis Pharmaceutical

Methods for high throughput screening of ion channel assays have been very limited to date. Although manual patch clamp evaluation of channel activity is the gold standard the technique can only handle a handful of compounds per week. Automated patch clamp methods promise to improve this performance dramatically. However, higher throughput methods such as Ca++ dyes, membrane potential dyes, and radioactive efflux assays all have their limitations. We will describe one new method, non radioactive Rb+ efflux measurements, that we are in the process of adding to our arsenal of techniques for studying potassium channels. We are measuring Rb+ efflux with Aurora Bioscience new atomic absorption instrument the ICR12000. We will present the theory behind these measurements and our initial testing of the system.

Presentation:  
Paul Hensley, President; Microplate Automation, Inc.; http://www.microplateautomation.com/

Paul Hensley, President of Microplate Automation, will present a technical overview of TipCharger?technology, the value proposition of the technology, and discuss product integration. TipCharger is a low temperature, atmospheric pressure plasma process that conditions and cleans pipetting devices. It is a replacement for solvent wash stations and disposable probe tip transfer stations on automated fluid handling equipment. The TipCharger apparatus removes organic substances from the treated surfaces of the pipet. Carryover is virtually unmeasurable. On contact with the plasma, organic substances are ionized into component molecules and form water, carbon dioxide and other similar compounds.

DON'T FORGET TO REGISTER TO INSURE THAT THERE IS ENOUGH FOOD AND SEATS. 
https://www.lab-robotics.org/member/meetings.asp?rid=1

Directions:
http://www.marriotthotels.com/SOSNJ

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