Luminex: Integrating Genomics & Protein Assays

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Luminex: Integrating Genomics & Protein Assays Clones Homebrew/Serology Cytokines Gene Expression xmap PhosphoProteins GE modulation (microrna) Transcription Factors

Luminex s xmap Liquid Bead Array Technology Dominic Andrada Life Science Research Business Manager dandrada@luminexcorp.com

Topics > What is Luminex > Benefits of using xmap > Applications > Backwards compatibility, forward thinking > Who is Luminex

What s Life Science Research Using?? > Technology platform basics > Industry advances in multiplex and parallel analysis > Test tubes > Microwell plate > Microarray > xmap Technology: Microsphere-based liquid array

What is xmap Technology? > Combination of multiple proven technologies > Color-coded 5.6 micron microspheres > Assay occurs on bead surface > Luminex LX200 Analyzer

Each Bead Type Is Assigned A Distinct Spectral Address Each bead maps to a specific region of the array based on the ratio of the two classification signals Red (660 nm) IR(infrared) (730 nm) Darker Conc. of Classfication dye 2 Conc. of Classification dye 1 Darker 100 color codes = 100 bead regions = 100 simultaneous tests in each well

How Does xmap Technology Work? Capture antibodies precoupled to unique bead set Cell supernate added to pooled bead sets, multiple cytokines captured Biotinylated antibodies bind to cytokines, detection via streptavidin-pe

xmap Technology Flow Cytometry Evolving > Assays are read using a compact microsphere analyzer > Analyzer samples well > Lasers excite fluorescent dyes - red laser for bead classification and green laser for assay result > Multiple readings for each microsphere set > Software reports results in real-time > Up to 9600 results read in 1 hour

xmap Technology Hardware Compact LX 200 Analyzer Reads 96-well plates, automated fluidics; wash, prime, shutdown Red laser for bead classification and green laser for analyte quantitation Software reports results in real-time Up to 9600 results read in 1 hour

xmap vs. Traditional Methods Measurement of cytokines in extracellular fluids ELISA Measurement of intracellular phosphoproteins Western Blot

Better interaction btw assay & analytes > The Beads > Microspheres are suspendable and easily multiplexed. > Liquid-Phase Kinetics allows greater interaction and equilibrium with analytes and sample matrix > Greater accessibility for epitope binding > Beads find their correct target

Benefits of Using xmap Technology - EFFICIENCY > Multiplex > Decrease sample volume requirements > Reduce assay reagent volume, expense & labor > Generate more information on interrelationships between analytes within a single sample > Bead-based array > More favorable liquid phase hybridization kinetics > Suitable for life science research and diagnostics > Potential to translate LS research to clinical diagnostic

Benefits of Using xmap Technology - SCALABILITY Each bead maps to a specific region of the array based on the ratio of the two classification signals Red (660 nm) IR(infrared) (730 nm) Darker Conc. of Classfication dye 2 Conc. of Classification dye 1 Darker Many analytes few samples Focused analytes many samples

Benefits of Using xmap Technology - FLEXIBILITY Applications

microrna: Bead-based hybridization vs. Planar array Method 2 Method 1 Nature Vol. 435, June 9, 2005 Jun Lu, et. al.

Luminex Assay Menus > FDA-cleared/IVD Clinical assays > More than 4 dozen diagnostic assays > Growing list of Research assays > Proteins > Gene Expression > microrna > Transcription Factors > Specialty Panels

xmap Microspheres Kits or Homebrew Beads: Carboxy SeroMAP LumAvidin FlexMAP MagPlex

Who is Luminex 1995 > Founded 1999 > First Instrument 1996 > First Patent 2000 > IPO (LMNX) 2005 > 50 Partners > Over 500 analytes > FDA cleared kits > 27 Patents issued > 52 patents pending 2008 5000 Placements FlexmiR microrna xponent RVP Panel FlexMAP3D Genomic Assays 1992 1994 1996 1998 2000 2002 2004 2006 2008

Luminex Business Model Multi-Partner model allows rapid development of the broadest assay menu > We license xmap technology to many kit developers > These partners provide market flexibility and multiple choices of vendors for end-users (Partners) (Customers) > Customers can also choose to develop custom-coupled assays with help from these partners or Luminex directly

Luminex in the Future > New Hardware: FlexMAP3D > 96 & 384-well > Magnetic Beads > Faster & Higher Thruput

Luminex: Integrating Genomics & Protein Assays Clones Homebrew/Serology Cytokines Gene Expression xmap PhosphoProteins GE modulation (microrna) Transcription Factors