Human exhaled aerosol droplet biomarker system and method
Patent Number: US9617582
Executive Summary:
General Description:
The invention presents an integrated system and method for detection of nonvolatile biomarkers in exhaled droplets of respiratory lining fluid from the distal lung. The proposed technique exceeds the sensitivity and reliability of the conventional exhaled breath condensate collection and assay systems, and includes various innovative components and techniques. The exhaled air collection is performed using the techniques that are currently applied to air pollution analyses. It provides the means for collection of particles with a range of sizes from nanodroplets through microdroplets, and a collection device for labile proteins from these particles. The collected particles are quantified using an optical device that determines the volume of respiratory fluid droplets generated by the patient. Therefore, direct standardization of the biomarker in lung lining fluid is achieved for each patient.
An additional technical advancement included in this invention overcomes the tedious manual effort usually associated with detection of multiple biomarkers using polymerase chain reaction. According to the present invention, antibodies are chemically immobilized into a microfluidic chip. Specifically, a multiplexed immunoquantitative polymerase chain reaction assay in a microfluidic chip is proposed, which simultaneously measures multiple targets in each sample with minimal hands-on effort. The invention has been applied to diagnostics of influenza, asthma, and chronic obstructive pulmonary disease in several cohorts of patients in clinical studies.
Future Directions:
Strengths:
Weaknesses:
Patent Status:
Publications:
1: Fabian P, McDevitt JJ, DeHaan WH, Fung RO, Cowling BJ, Chan KH, Leung GM, Milton DK. Influenza virus in human exhaled breath: an observational study. PLoS One. 2008 Jul 16;3(7):e2691. doi: 10.1371/journal.pone.0002691. PubMed PMID:18628983; PubMed Central PMCID: PMC2442192.
2: Fabian P, McDevitt JJ, Lee WM, Houseman EA, Milton DK. An optimized method to detect influenza virus and human rhinovirus from exhaled breath and the airborne environment. J Environ Monit. 2009 Feb;11(2):314-7. doi: 10.1039/b813520g. Epub 2008 Dec 1. PubMed PMID: 19212587; PubMed Central PMCID: PMC2840048.
3: Shorter JH, Nelson DD, Barry McManus J, Zahniser MS, Milton DK. Multicomponent Breath Analysis With Infrared Absorption Using Room-Temperature Quantum Cascade Lasers. IEEE Sens J. 2009 Dec 11;10(1):76-84. PubMed PMID: 20697459; PubMed Central PMCID: PMC2917122.
4: Fabian P, Brain J, Houseman EA, Gern J, Milton DK. Origin of exhaled breath particles from healthy and human rhinovirus-infected subjects. J Aerosol Med Pulm Drug Deliv. 2011 Jun;24(3):137-47. doi: 10.1089/jamp.2010.0815. Epub 2011 Mar 1. PubMed PMID: 21361786; PubMed Central PMCID: PMC3123971.
5: Sordillo JE, Webb T, Kwan D, Kamel J, Hoffman E, Milton DK, Gold DR. Allergen exposure modifies the relation of sensitization to fraction of exhaled nitric oxide levels in children at risk for allergy and asthma. J Allergy Clin Immunol. 2011 May;127(5):1165-72.e5. doi: 10.1016/j.jaci.2011.01.066. Epub 2011 Apr 3. PubMed PMID: 21463890; PubMed Central PMCID: PMC3137133.
6: Shorter JH, Nelson DD, McManus JB, Zahniser MS, Sama SR, Milton DK. Clinical study of multiple breath biomarkers of asthma and COPD (NO, CO(2), CO and N(2)O) by infrared laser spectroscopy. J Breath Res. 2011 Sep;5(3):037108. doi:10.1088/1752-7155/5/3/037108. Epub 2011 Jul 15. PubMed PMID: 21757803; PubMed Central PMCID: PMC3169766.
7: McKenzie JH, McDevitt JJ, Fabian MP, Hwang GM, Milton DK. Collection of aerosolized human cytokines using Teflon® filters. PLoS One. 2012;7(5):e35814. doi: 10.1371/journal.pone.0035814. Epub 2012 May 4. PubMed PMID: 22574123; PubMed Central PMCID: PMC3344827.
8: McDevitt JJ, Koutrakis P, Ferguson ST, Wolfson JM, Fabian MP, Martins M, Pantelic J, Milton DK. Development and Performance Evaluation of an Exhaled-Breath Bioaerosol Collector for Influenza Virus. Aerosol Sci Technol. 2013 Jan 1;47(4):444-451. Epub 2013 Jan 25. PubMed PMID: 23418400; PubMed Central PMCID: PMC3570155.
9: Milton DK, Fabian MP, Cowling BJ, Grantham ML, McDevitt JJ. Influenza virus aerosols in human exhaled breath: particle size, culturability, and effect of surgical masks. PLoS Pathog. 2013 Mar;9(3):e1003205. doi:10.1371/journal.ppat.1003205. Epub 2013 Mar 7. PubMed PMID: 23505369; PubMed Central PMCID: PMC3591312.
Inventor Bio: Donald K. Milton
http://sph.umd.edu/people/donald-milton
Executive Summary:
- Invention Type: Diagnostic and Device
- Patent Status: US grant, Active
- Patent Link: https://patents.google.com/patent/US9617582/
- Research Institute: University of Maryland
- Disease Focus: Lung disease
- Basis of Invention: The breathed-out air contains small droplets that carry a variety of proteins from the inner lining of the lung. When the lung disease occurs, the composition of the proteins is changed, and the specific nature of the change depends on the disease
- How it works: A specialized filter captures the droplets that the patient breathes out. The biomarkers (proteins) present in the captured material are analyzed by Immuno-quantitative polymerase chain reaction (IqPCR). The disease is diagnosed based on the biomarkers identified by IqPCR
- Lead Challenge Inventor: Donald K. Milton
- Inventors: Donald K. Milton, Ian M. White
- Development Stage: Phase I/II clinical studies
- Novelty:
- The small exhaled particles with intact nanodroplets containing disease biomarkers are collected using the air pollution analyses techniques, rather than using the condensation of water vapor
- Immuno-quantitative polymerase chain reaction (IqPCR) assay is utilized to detect as few as 100 molecules of biomarker
- Clinical Applications:
- Diagnosis and management of lung cancer, asthma, COPD, tuberculosis, influenza, and HIV/AIDS related respiratory infections
General Description:
The invention presents an integrated system and method for detection of nonvolatile biomarkers in exhaled droplets of respiratory lining fluid from the distal lung. The proposed technique exceeds the sensitivity and reliability of the conventional exhaled breath condensate collection and assay systems, and includes various innovative components and techniques. The exhaled air collection is performed using the techniques that are currently applied to air pollution analyses. It provides the means for collection of particles with a range of sizes from nanodroplets through microdroplets, and a collection device for labile proteins from these particles. The collected particles are quantified using an optical device that determines the volume of respiratory fluid droplets generated by the patient. Therefore, direct standardization of the biomarker in lung lining fluid is achieved for each patient.
An additional technical advancement included in this invention overcomes the tedious manual effort usually associated with detection of multiple biomarkers using polymerase chain reaction. According to the present invention, antibodies are chemically immobilized into a microfluidic chip. Specifically, a multiplexed immunoquantitative polymerase chain reaction assay in a microfluidic chip is proposed, which simultaneously measures multiple targets in each sample with minimal hands-on effort. The invention has been applied to diagnostics of influenza, asthma, and chronic obstructive pulmonary disease in several cohorts of patients in clinical studies.
Future Directions:
- Clinical trials for lung cancer diagnostics
Strengths:
- A comprehensive diagnostic device with well-developed and validated mechanical design and molecular biology techniques
Weaknesses:
- May be expensive for routine diagnostics
- Has not been validated on lung cancer patients
Patent Status:
- Filing date: 2013-09-04
- Publication date and Grant date: 2017-04-11
Publications:
1: Fabian P, McDevitt JJ, DeHaan WH, Fung RO, Cowling BJ, Chan KH, Leung GM, Milton DK. Influenza virus in human exhaled breath: an observational study. PLoS One. 2008 Jul 16;3(7):e2691. doi: 10.1371/journal.pone.0002691. PubMed PMID:18628983; PubMed Central PMCID: PMC2442192.
2: Fabian P, McDevitt JJ, Lee WM, Houseman EA, Milton DK. An optimized method to detect influenza virus and human rhinovirus from exhaled breath and the airborne environment. J Environ Monit. 2009 Feb;11(2):314-7. doi: 10.1039/b813520g. Epub 2008 Dec 1. PubMed PMID: 19212587; PubMed Central PMCID: PMC2840048.
3: Shorter JH, Nelson DD, Barry McManus J, Zahniser MS, Milton DK. Multicomponent Breath Analysis With Infrared Absorption Using Room-Temperature Quantum Cascade Lasers. IEEE Sens J. 2009 Dec 11;10(1):76-84. PubMed PMID: 20697459; PubMed Central PMCID: PMC2917122.
4: Fabian P, Brain J, Houseman EA, Gern J, Milton DK. Origin of exhaled breath particles from healthy and human rhinovirus-infected subjects. J Aerosol Med Pulm Drug Deliv. 2011 Jun;24(3):137-47. doi: 10.1089/jamp.2010.0815. Epub 2011 Mar 1. PubMed PMID: 21361786; PubMed Central PMCID: PMC3123971.
5: Sordillo JE, Webb T, Kwan D, Kamel J, Hoffman E, Milton DK, Gold DR. Allergen exposure modifies the relation of sensitization to fraction of exhaled nitric oxide levels in children at risk for allergy and asthma. J Allergy Clin Immunol. 2011 May;127(5):1165-72.e5. doi: 10.1016/j.jaci.2011.01.066. Epub 2011 Apr 3. PubMed PMID: 21463890; PubMed Central PMCID: PMC3137133.
6: Shorter JH, Nelson DD, McManus JB, Zahniser MS, Sama SR, Milton DK. Clinical study of multiple breath biomarkers of asthma and COPD (NO, CO(2), CO and N(2)O) by infrared laser spectroscopy. J Breath Res. 2011 Sep;5(3):037108. doi:10.1088/1752-7155/5/3/037108. Epub 2011 Jul 15. PubMed PMID: 21757803; PubMed Central PMCID: PMC3169766.
7: McKenzie JH, McDevitt JJ, Fabian MP, Hwang GM, Milton DK. Collection of aerosolized human cytokines using Teflon® filters. PLoS One. 2012;7(5):e35814. doi: 10.1371/journal.pone.0035814. Epub 2012 May 4. PubMed PMID: 22574123; PubMed Central PMCID: PMC3344827.
8: McDevitt JJ, Koutrakis P, Ferguson ST, Wolfson JM, Fabian MP, Martins M, Pantelic J, Milton DK. Development and Performance Evaluation of an Exhaled-Breath Bioaerosol Collector for Influenza Virus. Aerosol Sci Technol. 2013 Jan 1;47(4):444-451. Epub 2013 Jan 25. PubMed PMID: 23418400; PubMed Central PMCID: PMC3570155.
9: Milton DK, Fabian MP, Cowling BJ, Grantham ML, McDevitt JJ. Influenza virus aerosols in human exhaled breath: particle size, culturability, and effect of surgical masks. PLoS Pathog. 2013 Mar;9(3):e1003205. doi:10.1371/journal.ppat.1003205. Epub 2013 Mar 7. PubMed PMID: 23505369; PubMed Central PMCID: PMC3591312.
Inventor Bio: Donald K. Milton
http://sph.umd.edu/people/donald-milton
