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Dcninovatecpresentationfinal 101110124949-phpapp01

  1. 1. Complete, creative solutions for developers and manufacturers of rapid assays Complete, creative solutions for developers and manufacturers of rapid assays Seminar on Business and Innovation in Biotechnology INOVATEC Belo Horizonte, Brazil. October 5th, 2010
  2. 2. • Brief introduction to DCN • State of the art in applied rapid assay technology – Standard formats for standard applications – More advanced formats for quantification, high sensitivity and digital interpretation Agenda Complete, creative solutions for developers and manufacturers of rapid assays • Market Opportunities and Market Demands • Market Driven Innovation: Meeting market demand – Materials and reagents – Cassette and sample handler design – Reader development • System development case studies
  3. 3. Key Points 1. Rapid testing is a dynamically growing, readily accessible market with opportunities available in numerous market segments 2. Assays can be developed to handle almost any reasonable market condition, based on existing technology – Simple, low cost lateral flow assays for simple, qualitative low resolution applications – More complex, more expensive lateral flow systems for high Complete, creative solutions for developers and manufacturers of rapid assays – More complex, more expensive lateral flow systems for high sensitivity, more quantitative, difficult applications – These two types of assay are very different in how they are developed and manufactured – Other approaches to high performance rapid testing are possible, but with a much longer outlook. 3. All of the elements needed to manufacture high performance tests are available today, so while technological innovation is ongoing, we can also choose to innovate in terms of where we apply these technologies
  4. 4. Introduction to DCN • Based in Carlsbad, California • Founded in 2003 with the purpose of assisting companies in developing, manufacturing and commercializing rapid tests for any application. • Our primary platform technologies are lateral flow, flow Complete, creative solutions for developers and manufacturers of rapid assays through, microfluidic, dipstick (chemistry) and rapid ELISA, using visual, fluorescent or paramagnetic detection systems • We work in any market segment where these technologies can be applied
  5. 5. Primary Services and Products Rapid assay development and manufacturing From Feasibility and Prototyping to full Development and Manufacturing Transfer Education and training courses in rapid assay technology Learn how to develop and manufacture rapid tests Custom conjugation services protein – particle, protein-protein, small molecule etc Complete, creative solutions for developers and manufacturers of rapid assays Consulting on any aspect of rapid assay development and commercialization Products Materials starter kits Gold conjugation kits Latex conjugation kits Colloidal gold All related lateral flow materials Lateral flow reader systems: Fluorescent and visual
  6. 6. • Fully equipped for the manufacture of lateral flow, flow through, and other rapid assay formats • 8000 square feet, containing 3000 square feet of wet lab, housed in a class 10,000 capable cleanroom, and an associated controlled humidity room • ISO 9001:2008 compliant, with EN 13485 by end 2010 Facility and Personnel Complete, creative solutions for developers and manufacturers of rapid assays • ISO 9001:2008 compliant, with EN 13485 by end 2010 • Development staff of 14 industrially experienced assay development scientists Reagent Prep Area Main Lab (Class 10,000 capable cleanroom) Component Processing Area (Temperature / Humidity Controlled)
  7. 7. Our Markets Medical Diagnostics Veterinary Diagnostics Agricultural / Environmental Food Testing OTC/Consumer Our market is international and spread over multiple application areas. Complete, creative solutions for developers and manufacturers of rapid assays OTC/Consumer Pharmaceutical Biowarfare Law enforcement / Forensics Microbiology Nucleic Acid Testing Industrial Health and Safety Approximately 40% of our assay development programs are US- based, 40% are European and 20% are spread among Asia and Latin America
  8. 8. Representative Clients • Johnson and Johnson (UK) • Novozymes (Denmark) • Amic (Sweden) • ICP Bio (Australia) • Rockeby (Australia) • Government of Thailand NIH (Thailand) • Pacific Biotech (Thailand) • BioRad (US/EU) • Chiron Corporation (US) • Amgen (US) • Abbott (US) • Genzyme Diagnostics (US) • Becton Dickinson (US) • Church and Dwight (US) Complete, creative solutions for developers and manufacturers of rapid assays • Pacific Biotech (Thailand) • Span Diagnostics (India) • Daiichi Fine Chemical (Japan) • Elixir Biosciences (China) • MARS/Masterfoods (UK) • Laboratorio Silanes (Mexico) • Mintek (South Africa) • Biosensia (Ireland) • Church and Dwight (US) • 3M (US) • Whatman /GE (US / Europe) • Aspen BioPharma (US) • Genetic Testing Institute (GTI) (US) • Cibavision (US) • CDC (US) • NIH (US)
  9. 9. Simple Devices for Rapid Testing Standard Lateral Flow Strip Complete, creative solutions for developers and manufacturers of rapid assays Sample addition Conjugate solubilization Capillary flow; antibody binding Membrane clearing; optical reading Step 1 Step 2 Step 3 Step 4
  10. 10. Simple Devices for Rapid Testing Standard Flow – through device Complete, creative solutions for developers and manufacturers of rapid assays Sample addition Optional wash Conjugate addition; antibody binding Final wash; membrane clearing; optical reading
  11. 11. Evolution in Point of Care Assays Drivers for Innovation and Market Growth • Early POC devices were designed for qualitative applications • Earliest applications in the 1980s were pregnancy tests, growing in the 1990’s to include drugs of abuse and infectious disease tests • Market has grown steadily at a minimum CAGR of 10% since Complete, creative solutions for developers and manufacturers of rapid assays the late 1990’s • Market demand for tests in non-traditional areas has expanded to include quantitative, high sensitivity applications using difficult matrices, driving development • Intellectual property issues have driven technological innovation strongly in the past decade
  12. 12. Innovative Applications Complete, creative solutions for developers and manufacturers of rapid assays
  13. 13. Why lateral flow assays dominate the (non glucose) POC market • Ease of use: Minimal operator dependent steps and interpretation • Can handle small volumes of multiple sample types • Can have high sensitivity, specificity, good stability • Good Manufacturability and Scalability Complete, creative solutions for developers and manufacturers of rapid assays • Good Manufacturability and Scalability • Minimal market and user education required • Market presence and acceptance • Short development cycle – once reagents are developed, the timeline to manufacturing transfer and approval is short relative to other biotech products
  14. 14. • Sensitivity • Control of user interpretation • Reproducibility (CV)/ Quantification • Multiplexing is difficult Typical performance challenges in standard rapid assays Complete, creative solutions for developers and manufacturers of rapid assays
  15. 15. Application Requirements are Driving the Technology Forward •The requirements that the major rapid test markets are demanding include: • Speed to result: 2-10 minutes • Sensitivity: high picogram/ml to low nanogram/ml • Ability to handle difficult sample matrices • Quantification Complete, creative solutions for developers and manufacturers of rapid assays • Quantification • Multiplexing • Removal of interpretation issues • Control of data
  16. 16. Evolution in Rapid Test Technology: An Exercise in Market Driven Innovation Clients provide use inspired basic research Pasteur's Quadrant describes scientific research that both seeks fundamental understanding of scientific problems, and, at the same time, seeks to be eventually useful. Complete, creative solutions for developers and manufacturers of rapid assays Development in rapid diagnostic testing incorporates elements of use inspired basic research (application development) and pure applied research (technology development) DCN and partners provide the applied element
  17. 17. Bridging the Concept Gap DCN Complete, creative solutions for developers and manufacturers of rapid assays • Product Requirements Development • Concept Refinement • Feasibility and prototyping of all system elements • Product development and system integration • Transfer to manufacturing • Commercialization
  18. 18. Veterinary 9 Food and Beverage 1 Pharmceuticals 0.6 Envionmental 0.2 Water 0.2 Medical Diagnostics Veterinary Food and Beverage High Performance Allows for New Applications Rapid Test Market Penetration and Opportunities Complete, creative solutions for developers and manufacturers of rapid assays Medical Diagnostics 89 Percent of Market by Application Total Market 2007: USD $2.3 billion Total Market 2012: USD $$3.7 billion Pharmceuticals Envionmental Water
  19. 19. Major Rapid Test Market Opportunities 1. Medical Diagnostics 35% 20% 21% 13% 7% 4% % market 2007 Pregnancy Infectious diseases Cardiac markers Cholesterol/lipds Drugs of abuse Other •Growth opportunities exist in all medical segments •Growth will be driven by reimbursement in US and EU markets Complete, creative solutions for developers and manufacturers of rapid assays 20% 24% 27%26% 15% 5% 3% predicted market 2012 Pregnancy Infectious diseases Cardiac markers Cholesterol/lipds Drugs of abuse Other •Requires performance improvements in quantification, sensitivity, reproducibility and multiplexing
  20. 20. Major Rapid Test Market Opportunities 2. Veterinary Diagnostics • Market was $205 million (USD) in 2007 • 9% CAGR to 2012 predicts a $310 million market by 2012 • Highly fragmented, price sensitive market, split among small animals and livestock • Major segments are infectious disease testing, metabolic testing (cancer), diabetes and fertility/pregnancy Complete, creative solutions for developers and manufacturers of rapid assays •Small animal tests tend to be paid for by owners and are non- regulated for the most part. Large animal testing tends to be government mandated and is regulated •Growth driven by acceptance in large animal testing for infectious disease and fertility, and control of antibiotic use and in small animals by cancer and infectious disease testing •Requires performance improvements in quantification, sensitivity, reproducibility and multiplexing
  21. 21. Major Rapid Test Market Opportunities 3. Food and beverage testing • Worldwide market of about $30 million in 2007 •Expected CAGR of 15% predicts $60 million by 2012 •Market is under-penetrated by rapid tests due to the preponderance of microbiological testing (slow, traditional technology largely used) Complete, creative solutions for developers and manufacturers of rapid assays •Combinations of molecular and immunological testing may be important to this market •Growth will be driven by increased awareness of rapid formats coupled with sampling and performance improvements that can generate appropriate sensitivity •Requires performance improvements in quantification, sensitivity, reproducibility and multiplexing
  22. 22. Major Rapid Test Market Opportunities 4. Water Testing and Environmental Remediation • Worldwide market of about $5 million in 2007. • Expected CAGR of 4% predicts $6 million by 2012. • Highly regulated markets, with local or governmental agencies setting requirements for testing, often in association with AOAC (Association of Official Analytical Chemists International) • Most testing is performed in certified laboratories by approved methods Complete, creative solutions for developers and manufacturers of rapid assays methods • Rapid tests are typically performed as spot checks between mandated test periods • Also used for household and industrial testing of water quality in certain markets • Growth will be driven by increased awareness of rapid formats coupled with performance improvements •Requires performance improvements in quantification, sensitivity, reproducibility and multiplexing
  23. 23. • Option 1: Develop completely new platform technologies based on microfluidics and alternative detection technologies e.g. lab on a chip, Market – Driven Innovation Making test performance meet market demands Complete, creative solutions for developers and manufacturers of rapid assays • Option 2: Improve the existing technology platform
  24. 24. A System Approach • Rapid lateral flow and flow through assays can meet many of the market’s requirements, but a different approach to the assay is needed • High performing rapid assays need to be approached in the same way as high performance laboratory based systems • Key system elements: Complete, creative solutions for developers and manufacturers of rapid assays • Key system elements: 1) A properly developed assay made using 1) High quality materials 2) High quality reagents 3) Highly sensitive label for sensitive analyte detection 2) A well designed sample device and housing for the test 3) A controlled method for result interpretation 4) A well designed and controlled manufacturing process
  25. 25. 1. Assay Materials • Analytical procedures usually require multiple steps • Fluidics minimize number of steps in a test and increase ease of use • In most POC devices these steps are performed using treated porous matrices • Fluid flow in POC devices usually relies on capillary flow • Fluidic elements are usually designed to be disposable, and are Complete, creative solutions for developers and manufacturers of rapid assays • Fluidic elements are usually designed to be disposable, and are therefore made from low cost materials • Suppliers must be of the highest quality. Lowest cost materials will not support higher specified applications Test Line Control Line Particle Conjugate on conjugate pad Wick Sample Pad Nitrocellulose MembraneBacking
  26. 26. 1. Assay Reagents • Recognition is generally Ab-Ag or Ab-Ab based • Different characteristics are required for high performance in a lateral flow system than in an ELISA, for instance, due to the mechanics of the system: – The method chosen for screening is important Complete, creative solutions for developers and manufacturers of rapid assays • Important Parameters: – Specificity – Dissociation Constant (Kd) – High Affinity (High Binding Rate Constant (kon)) – Antibody Class (subtype: IgG1, IgG2A, IgG3, etc.) – Monoclonals typically preferred, but high purity polyclonals can also be used effectively 7777
  27. 27. 1. Assay Labels • Colloidal gold – Typically 20-40nm – Low cost, difficult to control passive conjugations, one color, relatively sensitive. Most common in qualitative applications • Latex: Colored, Fluorescent, Paramagnetic – Typically 100-300nm – Low cost, covalent conjugations, variety of chemistries, well Complete, creative solutions for developers and manufacturers of rapid assays – Low cost, covalent conjugations, variety of chemistries, well controlled, stable, variety of particles and labels. Most common for high sensitivity, quantitative applications (fluorescent, paramagnetic) • Others – Upconverting phosphors – Colloidal carbon – Direct labeling using fluorophores and enhancers
  28. 28. Commonly used fluorescent labels in POC Applications • We love fluorescence for high end applications! • DCN’s Experience: – Dark Red Labels (660-680nm) give good results due to lack of background fluorescence in biological fluids in that spectral area – Europium: Highly sensitive and commonly used due to broad Stokes’ shift and stability (300 – 625nm) Complete, creative solutions for developers and manufacturers of rapid assays Stokes’ shift and stability (300 – 625nm) – Direct labeling of antibodies using fluorescent dyes is also an option – sensitivity an issue – requires customization of system components.
  29. 29. 2. Readers The use of readers in lateral flow assay systems Detection of the signal generated by the reagents, translation into a numerical or alphanumerical output, and delivery of that output Use of readers allows for: Negative Positive 100% Visually Negative 100% Visually Positive Operator Dependent Negative or Positive Complete, creative solutions for developers and manufacturers of rapid assays Use of readers allows for: • Improved sensitivity • Reduction in user interpretation issues • Data capture, analysis and storage • Connectivity • Quantification • New applications Operator Dependent Negative or Positive Critical features: • Hardware • Algorithms for data processing, • Signal to noise reduction, • Measurement and data distribution • Calibration methods
  30. 30. Reader Supply Partners • Off-the-shelf readers are available that can be readily modified for any development program • DCN represents two suppliers: LRE and Qiagen Lake Constance (formerly ESE) – Provide a base platform from which to start – Have a history of development of this type of product Complete, creative solutions for developers and manufacturers of rapid assays – Have experience of regulatory and technical requirements – Can advise on important technical design considerations – May be faster to market – Generally have dedicated manufacturing capability • Readers can be custom designed by experienced designer such as Source Scientific, Polygenesis, Omnica (all DCN partners) – Slower and more expensive route in return for a custom solution
  31. 31. • Sensitive • Affordable • Simple to operate • Small, Mobile and Robust • Documented Results • Objective Results • Quantitative Results ESE Quant DCN’s Reader Partners: Handheld and Benchtop Readers Complete, creative solutions for developers and manufacturers of rapid assays • Quantitative Results DCN distributes both ESE and LRE readers DCN acts as the assay integration partner for both suppliers for customers worldwide LRE POC Reader 0 200 400 600 800 1000 1200 1400 0 200 400 600 800 1000 1200 1400 Data points Intensity[mV] Scan1 Scan2 Scan3 Scan4 Scan5 Scan6 Scan7 Scan8 Scan9 Scan10 Scan11 Scan12 Scan13 Scan14 Scan15 Scan16 Scan17 Scan18 Scan19 Scan20 Control Test
  32. 32. Comparison of gold and fluorescent labels LF Tuberculosis Fluorescent Assay • Based on a dark red dye (latex particle based) • Monoclonal sandwich assay • Compared directly to a colloidal gold method • Results read digitally using the ESE-Quant reader Complete, creative solutions for developers and manufacturers of rapid assays Dark Red Dye (Latex) ex 660 em 680 0 20 40 60 80 100 120 500 550 600 650 700 750 Wavelength (nm) Rel.Intensity Excitation Emission
  33. 33. LF Tuberculosis Fluorescent Assay Gold vs. Fluorescent Latex 6 8 10 12 300 400 500 600 700 PeakHeight(pixelunits) PeakHeight(mV) Sensitivity in TB Protein Assay: Gold vs Dark Red FlourescentLatex Complete, creative solutions for developers and manufacturers of rapid assays 0 2 4 0 100 200 0 0.1 0.5 0.75 1 1.25 5 PeakHeight(pixelunits) PeakHeight(mV) Concentration (ng/ml) FlourescentLatex Gold • Sensitivity greater than 0.2 ng/mL. • Using same antibody / antigen combination, the assay developed using a Dark Red fluorescent particle is >10 fold more sensitive than the assay developed using gold.
  34. 34. Anthrax Protective Antigen Assay • Assay for a recombinant Anthrax protein • Quantitative & qualitative results Complete, creative solutions for developers and manufacturers of rapid assays
  35. 35. 2 3 4 5 6 7 8 9 10 11 Signal/NoiseRatio Gold vs. Latex Comparison - Lateral Flow Anthrax Protective Antigen Assay Gold Fluorescent Anthrax Protective Antigen Results Complete, creative solutions for developers and manufacturers of rapid assays 0 1 2 1 10 100 1000 10000 Ng/mL rPA • Fluorescent assay sensitive to 2 ng/mL. Approximately 60- fold better sensitivity than comparable gold assay • Visual or reader based results
  36. 36. 3. Device Design The importance of device design in test performance • Cassette design plays an integral role in the performance of the assay • Early integration of cassettes with sample delivery elements and with strips is a critical element of the design process Complete, creative solutions for developers and manufacturers of rapid assays the design process • If a reader is to be used, design device with this in mind – off the shelf cassettes will rarely work • Novelty in approach can lead to major benefits e.g. separation of sample handling and conjugate mixing from the remainder of the device can significantly improve CV’s
  37. 37. The SymbioSysTm Program • Rapid diagnostics have historically had less emphasis on engineering than other industries. (ie. Medical Device) • Typical approach: Develop the strip, then find or try to design a cassette that works with it – highly inefficient and counter-productive. • With current demands for ease of use, precision, quantification, Complete, creative solutions for developers and manufacturers of rapid assays • With current demands for ease of use, precision, quantification, low cost of goods and reduced time to market, this method is no longer acceptable. • Concurrent engineering, assay development and user needs development is required to achieve this level of performance. • DCN and Symbient developed the SymbioSys program to address this demand.
  38. 38. DCN’s Design Partner: Symbient Product Development • Established in 2004. • Focus on development of single use diagnostic devices. • Product Types: – Lateral Flow Cartridges – Fluidic Cartridges with Integrated Functions – Sample Delivery Systems - pipettes, lancet devices, urine cups. Complete, creative solutions for developers and manufacturers of rapid assays urine cups. – Instrument Driven Systems
  39. 39. Services Provided • Mechanical Product Design • Industrial Design • 3D Solid Modeling (Solidworks) • Program Management • Precision Prototype Machining • Rapid Prototype Tooling and Molding • LIM Silicone Molding • Product Testing and Test Method Development. Complete, creative solutions for developers and manufacturers of rapid assays CONFIDENTIAL • Product Testing and Test Method Development. • Failure Modes and Effects Analysis (FMEA) • Process Development • Manufacturing Transition
  40. 40. Sample Handling Device and Cartridge Design • DCN and Symbient have • DCN and Symbient have developed various diagnostic cartridges including single test strip, multiple test strip and specialty housings. Complete, creative solutions for developers and manufacturers of rapid assays • DCN and Symbient have designed and developed several devices for collecting and processing samples for diagnostic testing. • Sample types include saliva, whole blood, fecal matter, urine, mucous and surfaces. • Methods include integrated buffers, mixing, filtering and metering.
  41. 41. Device Optical Reader Example: Sample handling in a quantitative agricultural test • Vertical flow assay • Rapid and quantifiable • Increased sensitivity with integrated pre-incubation • Particulate samples do not cause Complete, creative solutions for developers and manufacturers of rapid assays • Particulate samples do not cause flow problems 1. PRE-INCUBATION 2. FLOW THROUGH 3. RESULT
  42. 42. Alpha Amylase Detection in Wheat Hagberg Falling Numbers are determined by dropping a ball through a tube of ground wheat. The time it takes in seconds for it to reach the bottom correlates inversely to the amount of alpha-amylase in the wheat. High falling numbers indicate low concentrations of alpha-amylase, and low falling numbers indicate a high concentration of alpha-amylase. Falling Number Complete, creative solutions for developers and manufacturers of rapid assays 0 50 100 150 200 250 300 350 400 srw170 srw138 srw139 cbh2.7 cbh2.8 FallingNumber Wheat Sample Comparison of Values from Device and True Falling Number Values True FN DiagnostIQ Avg FN DiagnostIQ Avg [Amylase] Falling Number True FN avg amyl conc. St Dev %CV 304 1.54 6.4 1.77% 244 21.82 3.56 1.27% 223 36.02 16.8 4 6.77% 142 144.05 6.57 4.77%
  43. 43. Intellectual Property – Driven Innovation Rapiplex: A Novel Multiplex Rapid Assay Platform • RapiPlex: An integrated Point-of-Care in vitro diagnostic platform; • Unique intellectual property position • Multiplex, Quantitative and/or Qualitative, Versatile, Portable • Three integrating technologies to provide a robust platform • Fluidics Complete, creative solutions for developers and manufacturers of rapid assays • Fluidics • Immunodiagnostic assay systems • Integrated fluorescent reade
  44. 44. + Fluidic Chip • Proprietary fluidic strip & chip not constrained by lateral flow IP • Disposable oriented in the vertical position when an assay is run • No overlapping capillary materials – gaps between materials • Enables simultaneous multiplex analysis from a single sample • Defined individual channels in which sample flow can be controlled • Individual channel entry ports for each channel Complete, creative solutions for developers and manufacturers of rapid assays • Flow control zone ensures accurate and controlled flow in each channel • Injection molded – High Impact Polystyrene • Suitable for all assay formats and multiple biological fluid sample types – demonstrated on saliva, serum and whole blood to date
  45. 45. 400,000 500,000 600,000 700,000 800,000 Avg.PeakHeight(RFU) Biosensia Myoglobin Myoglobin assay Representative DCN Data Complete, creative solutions for developers and manufacturers of rapid assays 0 100,000 200,000 300,000 0 50 100 150 200 250 300 350 400 Avg.PeakHeight(RFU) Myoglobin Conc. (ng/ml) Error Bars – 95% Confidence
  46. 46. 150,000 200,000 250,000 300,000 Avg.PeakHeight(RFU) Biosensia Myoglobin – Low End Myoglobin assay Representative DCN Data Complete, creative solutions for developers and manufacturers of rapid assays 0 50,000 100,000 0 20 40 60 80 100 120 Avg.PeakHeight(RFU) Myoglobin Conc. (ng/ml) Error Bars – 95% Confidence
  47. 47. Myoglobin Conc. (ng/ml) 95% Confidence Limits 5 16,661 10,504 10 37,698 22,495 25 82,432 41,144 Myoglobin Assay Current DCN Sensitivity Data Complete, creative solutions for developers and manufacturers of rapid assays 25 82,432 41,144 50 145,149 106,220 100 295,043 183,477 350 764,314 483,319
  48. 48. Myoglobin Dilution Series (N=12) R² = 0.9985R² = 1 500,000 600,000 700,000 800,000 900,000 Avg.PeakHeight(RFU) Biosensia Myoglobin Before After Complete, creative solutions for developers and manufacturers of rapid assays 0 100,000 200,000 300,000 400,000 10 100 1000 10000 Avg.PeakHeight(RFU) log Myoglobin Conc. (ng/ml) After Poly. (Before) Poly. (After)
  49. 49. Summary • Solved major mechanical design issues • Assay CV’s controlled • Good Slope of Dose Response • Clinically relevant sensitivity (~5 ng/ml) Complete, creative solutions for developers and manufacturers of rapid assays
  50. 50. Manufacturing Process Design Membrane Conjugate Pad Dispense test and control lines Block membrane if necessary Pre-treat pad Dispense / dip conjugate Sample Pad Pre-treat pad Dry Dry Dry DryDry Complete, creative solutions for developers and manufacturers of rapid assays Assemble membrane, conjugate pad, sample pad, wick and backing into cards Cut cards into strips Assemble strips into cassettes Package cassettes
  51. 51. Process Control Batch Vs Continuous Processes XYZ 3050 dispenser with BioJets and AirJets Dispensing Dipping Drying Complete, creative solutions for developers and manufacturers of rapid assays BioJets and AirJets Clamshell benchtop laminator Manual Dip Tray CM 4000 Guillotine Cutter Lamination Cutting Forced Air Oven
  52. 52. Equipment and Processes Batch Vs Continuous Dispensing, Dipping, Drying Complete, creative solutions for developers and manufacturers of rapid assays Reel-to-reel web handling and dispensing platform Lamination, Cutting
  53. 53. In-line processing provides for greater process control and equivalent processing In line camera and bad part marking system on a BioDot RTR 4500 inline dispensing system Equipment and Processes Batch Vs Continuous Complete, creative solutions for developers and manufacturers of rapid assays 10 15 20 25 1 64 127 190 253 316 379 442 505 568 631 694 757 820 883 946 1009 1072 1135 1198 1261 1324 Pixels Mean Pixel Width Std Dev %CV 17.57 0.89 5.1 Output from a quality control camera system on a RTR 4500. Dispensing run over a 50m dispense length, 50mm/s, human IgG at 1mg/ml, 0.8ul/cm on nitrocellulose
  54. 54. All the elements of an integrated device are available over the counter • High quality test development • Reader development • Device development • Manufacturing equipment Complete, creative solutions for developers and manufacturers of rapid assays • A partnering approach is generally required to bring the specialized expertise and components into the assay
  55. 55. DCN’s Network Partners DCN’s Network of Affiliates is a group of closely knit companies that work together to provide all of the key elements necessary to design, develop manufacture and commercialize point of care diagnostic systems. Member companies include: • Manufacturing equipment suppliers Complete, creative solutions for developers and manufacturers of rapid assays • Cassette and device designers • High volume molding companies • Materials converters • Reader developers and manufacturers • Contract manufacturers • Platform technology suppliers
  56. 56. A reader based semi quantitative serological assay: Rapid Assay for anti-PF4:Heparin in human plasma (HIT) • Fluorescent, reader based, integrated RFID tag and novel cassette • ESEQuant fluorescent reader from Qiagen Lake Constance • Threshold assay, with Positive/Negative reporting • Predicate ELISA in market. Rapid assay must be as sensitive. • No known standards • Intended for the professional clinical laboratory market • Will be labeled as an IVD :developed under full design controls Complete, creative solutions for developers and manufacturers of rapid assays • Will be labeled as an IVD :developed under full design controls • Assay and device developed for large scale manufacturing
  57. 57. Major Challenges • Highly charged specific molecule • Total immunoglobulin assay imparts formatting requirement – sample and conjugate cannot mix prior to interaction with test line • High sensitivity needed • Lack of standards, so establishment of cutoff requires comparison to performance of predicate ELISA = Complete, creative solutions for developers and manufacturers of rapid assays comparison to performance of predicate ELISA = Requirement for high correlation to ELISA result. • Requirement for tight CVs • Integration of novel device design elements for IP generation • Predicate rapid assays in the field require multiple additions, timing steps and have poor user interface. The aim of the program is to generate a device without these attributes
  58. 58. Comparative results of lateral flow and ELISA data on all clinical samples 1000 1500 2000 2500 TestLinePeakHeight(RFU) OD vs Test Line Peak Height (All Data) OD vs TL Complete, creative solutions for developers and manufacturers of rapid assays 0 500 1000 0 0.5 1 1.5 2 2.5 3 3.5 4 TestLinePeakHeight(RFU) ELISA OD OD vs TL • A panel of 83 samples, characterized by predicate ELISA • All samples came from patients who had received heparin • Testing in triplicate / quadruplicate if adequate volume • Average values shown.
  59. 59. Cutoff: Comparative 0.400 0.500 0.600 0.700 TL/CL OD vs TL/CL (OD <1.0) OD vs TL/CL Complete, creative solutions for developers and manufacturers of rapid assays 0.000 0.100 0.200 0.300 0 0.2 0.4 0.6 0.8 1 TL/CL ELISA OD OD vs TL/CL 0.3 od Cutoff
  60. 60. OD = 0.3 ELISA TL/CL TL True positives 41 39 37 False Negative 0 2 4 True Negatives 42 42 42 False Positive 0 0 0 n 83 83 83 Collated Testing Results: ELISA vs LFIA using various cutoffs Complete, creative solutions for developers and manufacturers of rapid assays True Positive = Samples with OD > cutoff and detected as positive False negative= Samples with OD > cutoff and detected as negative True Negative= Samples with OD < cutoff and detected as negative False Positive= Samples with OD < cutoff and detected as positive n 83 83 83 PPV 100.00 100.00 100.00 NPV 100.00 95.45 91.30 Sensitivity 100.00 95.12 90.24 Specificity 100.00 100.00 100.00
  61. 61. Dilution Series: High positive Diluted in Negative R² = 0.9975 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 TL/CLRatio TL/CL Ratio Relative to ELISA OD, n=5 TL/CL ratio Poly. (TL/CL ratio) Complete, creative solutions for developers and manufacturers of rapid assays R² = 0.9989 -500 0 500 1000 1500 2000 2500 0.00 0.50 1.00 1.50 2.00 2.50 3.00 PeakHeight(rfu) ELISA OD Peak Test Line Height Relative to ELISA OD, n=5 5-14 serum TL Ht Poly. (5-14 serum TL Ht) 0.0 0.00 0.50 1.00 1.50 2.00 2.50 3.00 ELISA OD
  62. 62. Integrated System Development Using the DCN Network to develop complete assay systems • Project Management • System requirement definition • Product specification • Cassette design • Rapid prototyping • Mold design and development • Reader development • GUI design • Reagent Development • Assay design and development • Regulatory consulting • Quality system management Program Setup and Management System Development The client provides the innovation in reagents and application area DCN provides the rest Complete, creative solutions for developers and manufacturers of rapid assays • Mold design and development • Mechanical engineering design • GUI design • Assay design and development • Calibration system development • Documentation • Manufacturing process design • Quality system management • FMEA / Risk analysis • High volume plastic molding • Manufacturing equipment supply • Manufacturing facility setup • Scale-up and manufacturing transfer • Reagent manufacture • Gap manufacturing Commercialization • Market research • Board level advice • Networking to finance • Branding • Troubleshooting • Patent Landscaping Manufacturing Transfer
  63. 63. Key Points 1. Opportunities are available in numerous market segments for rapid assays with the right level of performance 2. Assays can be developed to handle almost any reasonable market condition, based on existing technology – Simple, low cost lateral flow assays for simple, qualitative low resolution applications – More complex, more expensive lateral flow systems for Complete, creative solutions for developers and manufacturers of rapid assays – More complex, more expensive lateral flow systems for high sensitivity, more quantitative, difficult applications – These two types of assay are very different in how they are developed and manufactured – Other approaches to high performance rapid testing are possible, but with a much longer outlook. 3. All of the elements needed to develop and manufacture high performance tests are available today
  64. 64. Acknowledgements • DCN – Roy Chung – Jerrold Loo – Wendy Mendiola – Michael Fiechtner • Symbient Product Development – John Zeis Complete, creative solutions for developers and manufacturers of rapid assays – John Zeis – Scott Castenon • Shara Rosen for marketing information • Thanks to GTI and Biosensia for use of data • Thanks to Cynthia Rocha, Camila Resende Duarte, Gislaine Silva, Regina Fernandes from SECTES and Christiane Contigli from CETEC and all who helped to coordinate my attendance at INOVATEC
  65. 65. www.dcndx.com Contact : Brendan O’Farrell at bofarrell@dcndx.com Diagnostic Consulting Network Inc 6354 Corte del Abeto, Suite B-C Contact Information Please email to receive copies of the slides Complete, creative solutions for developers and manufacturers of rapid assays 6354 Corte del Abeto, Suite B-C Carlsbad, CA 92011 Tel: 760-804-3886 Fax: 760-804-3866
  66. 66. • Quickly develop and commercialize rapid assays for any application • Experienced with fluorescent, visual (colloidal gold / latex) and paramagnetic labels – From concept to commercialized assay in under 12 months We can develop assays for you: Assay Development Services Complete, creative solutions for developers and manufacturers of rapid assays – From concept to commercialized assay in under 12 months – ELISA conversion to rapid assay formats in short timeframes – Reagent sourcing and development – Rapid prototyping – Feasibility studies – Complete development – Manufacturing transfer
  67. 67. • The plastic devices used in these assays can determine their ability to perform in demanding applications and need to be custom developed in virtually any high performance application • DCN partners with product designers, such as Symbient Product Development and molding companies such as HK Plastics to integrate We can design and develop your plastic devices: Cartridge and Device Design Service Complete, creative solutions for developers and manufacturers of rapid assays new cassettes and novel device designs into programs for clients • Partners can be contracted through DCN, and DCN will manage the entire development program on behalf of the client
  68. 68. • DCN partners with the major suppliers of readers to the point of care marketplace to provide reader solutions for our customers. • We can provide readers for: – Visual labels – Fluorescent labels – Paramagnetic labels We can help you develop reader systems for your assays Reader Development and Integration Complete, creative solutions for developers and manufacturers of rapid assays – Paramagnetic labels • Our primary partners for off the shelf and custom reader supply are QLC (formerly ESE) and LRE (Esterline), and we distribute for both companies in the US. • We also work with custom reader design and development houses including Source Scientific and Polygenesis.
  69. 69. • DCN can develop and commercialize all aspects of a point of care diagnostics platform, through our in- house capabilities and our Network of Affiliates We can design and develop your complete assay system Integrated System Design and Development Cartridge top housing RF id tag Cartridge bottom housingStrip and Assay Complete, creative solutions for developers and manufacturers of rapid assays • Custom design and integration for high performance assays our specialty Desiccant
  70. 70. We can connect you to all of the best suppliers: The Network of Affiliates DCN’s Network of Affiliates is a group of closely knit companies that work together to provide all of the key elements necessary to design, develop manufacture and commercialize point of care diagnostic systems. Member companies include: • Manufacturing equipment suppliers Complete, creative solutions for developers and manufacturers of rapid assays • Cassette and device designers • High volume molding companies • Materials converters • Reader developers and manufacturers • Contract manufacturers • Platform technology suppliers
  71. 71. • Learn how to develop and manufacture lateral flow assays • Customized one-on-one courses held at DCN’s core facility in Carlsbad, California • Scheduled on demand • Duration 3 – 5 days We can teach you to develop and make your own assays Education and Training Services Complete, creative solutions for developers and manufacturers of rapid assays • Theoretical and practical elements dealt with • Bring your reagents and optimize an assay during the course
  72. 72. We can help with any aspect of commercialization Consulting Services DCN has over 20 consultants on staff that can provide a wide range of services related to the point of need diagnostics or detection business: • Regulatory Affairs • Project management • Risk assessment • Intellectual property landscaping Complete, creative solutions for developers and manufacturers of rapid assays • Intellectual property landscaping • Manufacturing process development • Assay troubleshooting • Quality system establishment for diagnostic companies • Device design • Facility setup • Equipment specification and procurement • Market evaluation • Technology evaluation
  73. 73. • DCN’s facility is completely equipped for manufacture of lateral flow, flow through, and other format point of need assays • The facility: – 8000 square feet, containing 3000 square feet of wet lab, housed in a class 10,000 capable cleanroom, and an associated controlled humidity room – Currently ISO 9001:2008 compliant and will complete EN 13485 by end Q4 2010 • Manufacture non-FDA regulated product for several clients in the US and Europe, and We can manufacture your tests for you Contract Manufacturing Complete, creative solutions for developers and manufacturers of rapid assays components and reagents for clients in the US and outside • DCN partners with several dedicated contract manufacturers to whom it can transfer product for manufacturing. Reagent Prep Area Main Lab (Class 10,000 capable cleanroom) Component Processing Area (Temperature / Humidity Controlled)
  74. 74. • DCN provides everything needed to develop or manufacture lateral flow or flow through assays – Colloidal gold, colored and fluorescent latex beads and conjugates – Assay materials: membranes, pads, backing, cassettes wicks, etc – Antibodies and antigens – Small molecule conjugates We can supply you with key components Materials, Reagents and Kits Complete, creative solutions for developers and manufacturers of rapid assays – Small molecule conjugates – Test strip readers: Handheld and benchtop • Kits of materials and reagents – Colloidal gold – Material Starter Kit – Colloidal gold conjugation kit – Latex conjugation kit – Gold conjugation kit
  75. 75. We can make the critical reagents for you Custom Conjugate Development and Manufacture • Expertise in covalent conjugations – Protein / peptide /small molecule –Particle • Colored, fluorescent or paramagnetic latex, gold, quantum dots, etc. – Protein – Protein Streptavidin Bead #2 2.5 Strep #2 (1X) Complete, creative solutions for developers and manufacturers of rapid assays • Full optimization process • Full QC including binding assays where reagents are available • For diagnostics or separations (magnetic latex) • Small to full prep scale -0.5 0 0.5 1 1.5 2 0 5 10 15 20 25 ng Biotin OD Strep #2 (1X) Strep #2 (10X) Strep #2 (20X) Merck Strep REC/1.0um Candidat 2 Merck Strep C-REC/ 1.0um Candidat 4 Merck Strep TREC/1.0um Candidat 4 Merck Strep 0.2umCandidat 1
  76. 76. • Dr Roy Chung: Over 25 years of assay development and manufacturing experience. Previously Lead Scientist at Nanogen • Dr Hans Boehringer: Over 20 years of assay development and manufacturing experience. Most recently Scientific Director at Genzyme Diagnostics • Dr Brendan O’Farrell: 20 years of assay development, manufacturing, equipment and manufacturing process development. Previously VP of Technology Development at BioDot Inc and Director of R&D and Manufacturing at SDS Inc. Our People Complete, creative solutions for developers and manufacturers of rapid assays at SDS Inc. • Michael Surmanian: 20 years of biotech technology development experience. Formerly President of BioDot Inc. • Assay Development Team of 10 scientists, all of whom worked for major human and veterinary diagnostic companies. Minimum of 5 years of experience Consulting Group.: Over 25 industry experts available. Examples: • Dr Judith Howard: Ex VP of Quality, Regulatory at Inverness Medical Innovations • Dr David Boudreau: Ex Director of Operations, Nanogen Inc • Michael Weinholt: President of Regulatory Consulting Inc
  77. 77. Our Strengths • Expertise in reagent development, assay development, device design and reader integration • Experience in all aspects of system development, regulation, commercialization and manufacturing • Modular menu of services, allowing for flexible project execution – Complete, creative solutions for developers and manufacturers of rapid assays • Modular menu of services, allowing for flexible project execution – select only the services you need • Parallel development strategies for device and assay development – science and engineering combined • Networked to all of the partners you will need to get to market • Full design controls and documentation. • Full support for registration activities • Speed to commercialization
  78. 78. 1) Assay • Materials for fluid movement and reaction matrices • Biological recognition reagents (antibodies/antigens) • Signal generation reagents (labels) 2) Reader system Challenges in Designing and Manufacturing High Performance POC Assays Complete, creative solutions for developers and manufacturers of rapid assays • Technology • Supplier 3) Device • Sample collection and processing • Conjugate handling and mixing • Cassettes 4) Manufacturing process design
  79. 79. Market Penetration by Application 25 30 35 40 45 % of Available Market, 2007 (compared to lab based immunoassays that have a rapid segment) Complete, creative solutions for developers and manufacturers of rapid assays 0 5 10 15 20 Medical Diagnostics Veterinary Food and Beverage Pharmceuticals Envionmental Water
  80. 80. 60 70 80 90 Percentage of Available Market by Geographical Region Market Penetration by Geographical Region Complete, creative solutions for developers and manufacturers of rapid assays 0 10 20 30 40 50 US EU ROW
  81. 81. • Sensitivity: • Fluorescence based tests can be more sensitive than colorimetric • Higher sensitivity allows new applications and markets • Control • Fluorescent assays are based on latex particles which can be Why we like fluorescent labels for reader based applications Complete, creative solutions for developers and manufacturers of rapid assays coupled covalently. Colloidal gold conjugation is passive. • This allows for better control of the conjugates and improved CV‘s • Availability • High quality, high performance beads available • Multiple sizes, chemistries • Availability of off-the shelf readers • Makes development easy and low cost • Makes fluorescence a comparatively open system
  82. 82. Device Design Trends and Implications • IP considerations have driven a trend towards novel device design: – clear pathways to designing non-infringing devices exist • For many applications, sample handling, sample preparation, extraction and delivery to devices need to be performed Complete, creative solutions for developers and manufacturers of rapid assays – E.g. Tests for agriculture, biowarfare, food pathogen testing, environmental testing involve handling difficult samples, treating them and delivering them to devices in difficult field conditions. – Devices can be designed and integrated to cassettes to perform these functions
  83. 83. Myoglobin Conc. (ng/ml) n Avg Peak Height CV 0 15 0 na 5 15 13,583 11.3% 10 15 30,097 12.6% Myoglobin assay data Representative DCN Data Complete, creative solutions for developers and manufacturers of rapid assays 10 15 30,097 12.6% 25 15 61,788 16.7% 50 15 125,685 7.7% 100 15 239,260 11.7% 350 15 623,817 11.3%
  84. 84. Prototyping Capabilities CNC Machining Steriolithography (SLA) Complete, creative solutions for developers and manufacturers of rapid assays CNC Machining Prototype Mold Fabrication Injection Molding Steriolithography (SLA)
  85. 85. Assembly and Testing Complete, creative solutions for developers and manufacturers of rapid assays
  86. 86. Myo Conc. (ng/ml) N Avg. TL pk ht TL %CV 25 12 64,948 11.0% 50 12 118,659 5.9% Myoglobin Dilution Series (N=12) Complete, creative solutions for developers and manufacturers of rapid assays 100 12 229,002 10.4% 250 12 457,563 4.1% 500 12 749,168 6.9%
  87. 87. Overall Conclusion • Doing so requires careful design of all aspects of the product and the integration of high quality fluorescent label, cassette and reader technologies • The design and development of products of this nature is a highly specialized task • That is where a company like DCN, with specialist knowledge and experience, comes into the picture • How can we help you?........ Complete, creative solutions for developers and manufacturers of rapid assays • How can we help you?........

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