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1. (WO2019060980) FLUORESCENT ENVELOPED VIRAL PARTICLES AS STANDARDS FOR NANOSALE FLOW CYTOMETRY
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Pub. No.: WO/2019/060980 International Application No.: PCT/CA2018/000080
Publication Date: 04.04.2019 International Filing Date: 27.04.2018
IPC:
C12N 15/62 (2006.01) ,C07K 14/15 (2006.01) ,C07K 19/00 (2006.01) ,C12N 15/46 (2006.01) ,C12N 15/867 (2006.01) ,C12N 15/10 (2006.01) ,C12N 7/01 (2006.01) ,C12Q 1/6897 (2018.01) ,C12Q 1/70 (2006.01) ,G01N 15/10 (2006.01) ,G01N 33/48 (2006.01) ,G01N 33/52 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
62
DNA sequences coding for fusion proteins
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
005
from viruses
08
RNA viruses
15
Retroviridae, e.g. bovine leukaemia virus, feline leukaemia virus, human T-cell leukaemia-lymphoma virus
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
19
Hybrid peptides
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
31
Genes encoding microbial proteins, e.g. enterotoxins
33
Genes encoding viral proteins
40
Proteins from RNA viruses, e.g. flaviviruses
46
Reoviridae, e.g. rotavirus, bluetongue virus, Colorado tick fever virus
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79
Vectors or expression systems specially adapted for eukaryotic hosts
85
for animal cells
86
Viral vectors
867
Retroviral vectors
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
10
Processes for the isolation, preparation or purification of DNA or RNA
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
7
Viruses, e.g. bacteriophages; Compositions thereof; Preparation or purification thereof
01
Viruses, e.g. bacteriophages, modified by introduction of foreign genetic material
[IPC code unknown for C12Q 1/6897]
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
70
involving virus or bacteriophage
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
15
Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
10
Investigating individual particles
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
50
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
52
Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper
Applicants:
UNIVERSITY OF OTTAWA [CA/CA]; 3042-800 King Edward Ave Ottawa, Ontario K1N 6N5, CA
Inventors:
LANGLOIS, Marc-André; CA
TANG, Vera; CA
RENNER, Tyler Milston; CA
Agent:
BOOCOCK, R. Graeme; Borden Ladner Gervais LLP 100 Queen Street, Suite 1300 Ottawa, Ontario K1P 1J9, CA
Priority Data:
62/563,95727.09.2017US
Title (EN) FLUORESCENT ENVELOPED VIRAL PARTICLES AS STANDARDS FOR NANOSALE FLOW CYTOMETRY
(FR) PARTICULES VIRALES ENVELOPPÉES FLUORESCENTES EN TANT QUE RÉFÉRENCES POUR LA CYTOMÉTRIE EN FLUX NANOMÉTRIQUE
Abstract:
(EN) Described herein are uses of fluorescent enveloped virus particles as standards in nanoscale flow cytometry applications. The virus particles comprise a fluorescent dye or a fluorescent protein. A standard ladder comprising a plurality of fluorescent enveloped virus particles of different sizes is also provided. The standards may also comprise marker(s) of interest, and may be used as controls for detection of other viruses or extracellular vesicle, e.g. in diagnostic applications. Methods of producing controls for such applications are provided, including those having desired profiles. The controls may be used for enumeration of markers on microparticles (e.g. extracellular vesicles or viruses). Also described is a modified gammaretrovirus comprising a mutation that abolishes expression of the viral glyco-Gag protein, and having a fluorescent protein inserted in-frame into the proline-rich region (PRR) of the viral env protein. The gammaretrovirus may be M- MLV bearing a mutation that abolishes expression of the glyco-Gal protein, gPr80.
(FR) L'invention concerne des utilisations de particules virales enveloppées fluorescentes en tant que références dans des applications de cytométrie en flux nanométrique. Les particules virales comprennent un colorant fluorescent ou une protéine fluorescente. L'invention concerne également une échelle de référence comprenant une pluralité de particules virales enveloppées fluorescentes de différentes tailles. Les références peuvent également comprendre un ou plusieurs marqueurs d'intérêt et peuvent être utilisées en tant que témoins pour la détection d'autres virus ou de vésicules extracellulaires, par exemple dans des applications de diagnostic. L'invention concerne également des procédés de production de témoins pour de telles applications, y compris ceux présentant des profils souhaités. Les témoins peuvent être utilisés pour l'énumération de marqueurs sur des microparticules (par exemple des vésicules extracellulaires ou des virus). L'invention concerne également un gammaretrovirus modifié comprenant une mutation qui annule l'expression de la protéine virale glyco-Gag et présentant une protéine fluorescente insérée dans le cadre dans la région riche en proline (PRR) de la protéine env virale. Le gammaretrovirus peut être M-MLV portant une mutation qui annule l'expression de la protéine glyco-Gal, gPr80.
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: English (EN)
Filing Language: English (EN)