Conference Program - ABRF 2011 - Association of Biomolecular ...

Research GroupPresentation Abstracts(R6b) Conclusions from the MIRG 2010 BenchmarkStudy: Molecular Interactions in a Three ComponentSystem and Presentation of 2011 Survey Results onLabel-Free TechnologiesA.P. Yamniuk 1 , S.P. Yadav 5 , S. Bergqvist 2 , M.L. Doyle 1 ,E. Eisenstein 3 , M.K. Robinson 4 , T. Neubert 61Bristol-Myers Squibb, Princeton, NJ, United States; 2 Pfizer, LaJolla, CA, United States; 3 University Maryland BiotechnologyInstitute, Baltimore, MA, United States; 4 Fox Chase CancerCenter, Philadelphia, PA, United States; 5 Cleveland ClinicFoundation, Cleveland, OH, United States; 6 New YorkUniversity School of Medicine, New York, NY, United StatesCharacterizing the assembly of multi-protein complexes and thecompetition between multiple protein ligands for a given target arecommon challenges faced by core facilities. The MIRG2010 Benchmarkstudy was designed to assess participants’ ability to correctly describethe interactions between two protein ligands and their target proteinusing primarily biosensor technologies such as surface plasmonresonance. Participants were provided with microgram quantities ofthree proteins (A, B and C) and asked to determine if a ternary A-B-Ccomplex can form, or if ligands B and C bind competitively to proteinA. This presentation will summarize the conclusions from the 2010Benchmark Study, and provide perspective on the potential for futureapplication of this system as a reference standard for quantitativecharacterization of protein-protein interactions using biosensortechnologies. The field of label-free biophysical technologies likesurface plasmon resonance (SPR) and isothermal calorimetry (ITC) arebecoming indispensable in translational research and in the discoveryphase of biotherapeutics. Investigators are much more aware about thedevelopments in biomolecular interaction analysis using SPR and ITCand usefulness of these technologies in designing better drugs basedon biomolecules and vaccines. The Molecular Interaction ResearchGroup (MIRG) of ABRF has conducted an on-line survey to capture therecent explosive developments in these technologies. The survey wastargeted to both academia and pharmaceutical industry and the surveydata will be presented during the meeting.(R7) Light Microscopy Research Group (LMRG)(R7-1) Point Spread Functions, Spectral Calibration,and BeyondR. Stack, R. ColeWadsworth Center, New York State Department of Health,Albany, NY, United StatesModern light microscopes are highly evolved opto-electronicmechanicaldevices. Establishing instrument performance is crucial inensuring that reliable and accurate images can be acquired. Imagers, aswell as granting agencies, need to be confident that data collected willbe uniform and quantifiable both temporally and from instrument toinstrument. Last year, in phase-one of our world-wide research studyon instrument performance, we successfully concentrated our effortson three image-based tests: long and short term stability of illuminationsources, uniformity of field illumination, and co-registration acrossvarious wavelengths. A manuscript summarizing the phase-one studyhas been submitted to and accepted by Microscopy & Microanalysis,one of the highest rated imaging journals. In the coming year, phasetwoof our instrument performance study will focus on determining thefollowing: the point spread function of an imaging system, the system’sspectral separation ability and the spectral calibration and resolutionof the detection system. As with the phase-one study, the goal of thisstudy will be neither to compare the performance of different brandsof instruments, nor to ascertain which brand had better performancein a given area. Instead, the goal of our proposed phase-two study iscontinue to focus on determining the current state of modern imagingsystems through straightforward, efficient and robust tests. These testswill aid imagers in the early detection of system problems. Moreover,these tests will continue to help define relative standards that will assistboth core personnel and imagers in maintaining their instruments inoptimal operating conditions.(R7-2) Deconvolution: Core Concepts, Algorithmsand Advanced IssuesB. Northan, N. BeaversMedia Cybernetics, Guilderland, NY, United StatesDeconvolution is a computational technique used to remove blurfrom images. In this presentation image formation in the microscopewill be reviewed and it will be explained what deconvolution doesand why deconvolution is needed as a post acquisition processingstep. The concept of Point Spread Function (PSF) will be introduced.Several approaches to deconvolution and deblurring exist from simplesubtractive methods to complex iterative approaches. The majorapproaches will be examined. It will be explained why in the presenceof noise a statistical approach is required. Blind deconvolution will beintroduced. The concept of Point Spread Function will be examinedin detail. The effect of microscope modality, lens parameters, andspecimen parameters on PSF shape will be discussed. Theoretical PSFcalculation and PSF measurement using beads will be reviewed withexamples. It will be explained why theoretical and measured PSFs candiffer from the true system PSF. Blind deconvolution will be furtherexamined as a tool to deal with the uncertainty of the true form ofthe PSF. Practical guidelines for data acquisition will be reviewed. Therelationship between sampling rate and quality of deconvolutionresults will be explained.58 • ABRF 2011 — Technologies to Enable Personalized Medicine