Use of focused light scattering techniques in biological applications
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
G01N-033/569
G01N-015/02
G01N-015/14
G01N-033/574
G01N-015/00
출원번호
US-0283600
(2016-10-03)
등록번호
US-9683920
(2017-06-20)
발명자
/ 주소
Gabriel, Don
출원인 / 주소
AIDIA INVESTMENT GROUP, INC.
대리인 / 주소
Andrews Kurth Kenyon LLP
인용정보
피인용 횟수 :
0인용 특허 :
9
초록▼
Methods for using focused light scattering techniques for the optical sensing of biological particles suspended in a liquid medium are disclosed. The optical sensing enables one to characterize particles size and/or distribution in a given sample. This, in turn, allows one to identify the biological
Methods for using focused light scattering techniques for the optical sensing of biological particles suspended in a liquid medium are disclosed. The optical sensing enables one to characterize particles size and/or distribution in a given sample. This, in turn, allows one to identify the biological particles, determine their relative particle density, detect particle shedding, and identify particle aggregation. The methods are also useful in screening and optimizing drug candidates, evaluating the efficacy and dosage levels of such drugs, and in personalized medicine applications.
대표청구항▼
1. A method of determining whether a putative therapeutic agent binds to, and forms a complex with, a receptor on a biological particle of interest, comprising: a) obtaining a spectra showing particle size and distribution for a sample medium which includes a biological particle of interest with a r
1. A method of determining whether a putative therapeutic agent binds to, and forms a complex with, a receptor on a biological particle of interest, comprising: a) obtaining a spectra showing particle size and distribution for a sample medium which includes a biological particle of interest with a receptor to which a putative therapeutic agent will bind, by passing the sample medium past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, where particle size is reflected by the location of one or more peaks on the spectrum, and the number of particles is reflected by the area under the curve of the peak,b) incubating the sample medium with a putative therapeutic agent,c) obtaining a second spectra showing particle size and distribution on the incubated sample medium using focused light scattering techniques, andd) determining whether the particle size and distribution has been altered by the incubation of the putative therapeutic agent, wherein a change in the particle size and/or distribution is indicative of a complex formed of the putative therapeutic agent and the biological particle,wherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, and when the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both. 2. The method of claim 1, wherein the biological particle is selected from the group consisting of tumor cells, red blood cells, white blood cells, granulocytes, platelets, monocytes, neutrophils, lymphocytes, bacteria, viruses, and fungi. 3. The method of claim 1, wherein the biological particle is 0.1 μm to about 20 μm. 4. The method of claim 1, wherein the sample medium comprises a fluid selected from the group consisting of blood, blood products, water, cerebrospinal fluid, ascites, pleural fluid, and synovial fluid. 5. The method of claim 1, wherein the putative therapeutic agent is complexed to a microparticle, such that there is a measurable size difference between the uncomplexed biological particle prior to incubation with the putative therapeutic agent and the particle when complexed to the therapeutic agent after the particle and therapeutic agent are incubated. 6. A method for determining whether a biological particle will form a complex with a known therapeutic agent, comprising: a) obtaining a spectra showing particle size and distribution for a sample medium which includes a biological particle of interest with a receptor to which a known therapeutic agent may or may not bind, by passing the sample medium past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, where particle size is reflected by the location of one or more peaks on the spectrum, and the number of particles is reflected by the area under the curve of the peak,b) incubating the sample medium with a known therapeutic agent,c) obtaining a second spectra showing particle size and distribution on the incubated sample medium using focused light scattering techniques, andd) determining whether the particle size and distribution has been altered by the incubation of the known therapeutic agent, a change in the particle size and/or distribution is indicative of a complex formed of the known therapeutic agent and the biological particle,wherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, and when the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both. 7. The method of claim 6, wherein the biological particle is selected from the group consisting of tumor cells, red blood cells, white blood cells, granulocytes, platelets, monocytes, neutrophils, lymphocytes, bacteria, viruses, and fungi. 8. The method of claim 6, wherein the biological particle has a size in the range of from about 0.1 μm to about 20 μm. 9. The method of claim 6, wherein the sample medium comprises a biological fluid from the group consisting of blood, blood products, water, cerebrospinal fluid, ascites, pleural fluid, and synovial fluid. 10. A method for determining the minimum amount of a therapeutic agent required to effectively bind a known cell, microbe or virus comprising: a) generating a first spectrum showing particle size and distribution for a sample medium which includes a known cell, microbe, or virus, by passing the sample medium past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,using focused light scattering to prepare a spectrum showing particle size distribution from within the sample medium, where particle size is reflected by the location of one or more peaks on the spectrum, and the number of cell, microbe, or virus particles is reflected by the area under the curve of the peak;b) incubating a first concentration of a therapeutic agent with the known cell, microbe or virus;c) generating a second spectrum showing particle size and distribution using focused light scattering of the combination of the therapeutic agent and the known cell, microbe or virus; andd) comparing the first and second spectra, wherein a change in the particle size and/or distribution is indicative of binding and complex formed between the therapeutic agent and the cell, microbe or virus, and wherein binding is indicative of inhibition of the known cell, microbe or virus;e) repeating steps a-d with varying amounts of the therapeutic agent; andf) comparing the spectra to determine the minimum amount of therapeutic agent required to effectively bind the known cell, microbe or virus,wherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, and when the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both. 11. The method of claim 10, wherein the therapeutic agent is an antibody. 12. The method of claim 10, wherein the known cell, microbe or virus is a cell selected from the group consisting of tumor cells, red blood cells, white blood cells, granulocytes, platelets, monocytes, neutrophils, and lymphocytes. 13. The method of claim 10, wherein the known cell is a cancer cell and the therapeutic agent is effective in treating cancer. 14. The method of claim 10, wherein the known cell, microbe or virus is a microbe selected from the group consisting of bacteria and fungi. 15. The method of claim 14, wherein the known microbe is a bacteria and the therapeutic agent is effective in treating bacterial infection. 16. The method of claim 10, wherein the known cell, microbe or virus is a virus. 17. The method of claim 10, wherein the minimum amount of a therapeutic agent required to effectively bind a known cell, microbe or virus is correlated to an effective therapeutic dosage by determining a concentration of the therapeutic agent that corresponds to the minimum amount of the therapeutic agent, and then an effective dosage is calculated based on the dosage of the therapeutic agent required to provide the concentration. 18. A method of detecting particle shedding resulting from a cell interaction comprising: a) generating a first spectrum showing particle size and distribution for a sample medium which includes a known cell, by passing the sample medium past a focused light beam formed by passing a collimated light beam through an acceptance aperture to narrow the width of the beam,using focused light scattering techniques to prepare a spectrum showing particle size distribution from within the sample medium, where particle size is reflected by the location of one or more peaks on the spectrum, and the number of particles is reflected by the area under the curve of the peak,b) incubating a potential therapeutic agent with the known cell in the sample medium, wherein the potential therapeutic agent is a microparticle;c) generating a second spectrum showing particle size and distribution, using focused light scattering techniques, for the incubated sample medium;d) comparing the first and second spectra, wherein the presence of, or increase in particle density of, particles of a size less than those of the original known cell is indicative of particle shedding,wherein said focused light scattering techniques involve sensing single particles suspended in a sample medium when the sample medium is passed through a focused beam of light, such that, when the focused beam of light passes through the sample medium without being scattered by a particle, the beam passes on to a photodetector and the intensity is measured,wherein the focused beam is of a size such that a particle in the size range of 0.1 to 10 μm is sufficient to block all of the beam, or a significant enough part of the beam, so that the particle size can be measured, and when the beam is scattered, in whole or in part, by a particle, the intensity of the beam hitting the photodetector is altered, and the particle size and/or concentration are calculated using light-extinction, light-scattering detection, or both.
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