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1. (WO2018002125) LONG LIFETIME ALPHA-HEMOLYSIN NANOPORES
Latest bibliographic data on file with the International BureauSubmit observation

Pub. No.: WO/2018/002125 International Application No.: PCT/EP2017/065972
Publication Date: 04.01.2018 International Filing Date: 28.06.2017
IPC:
C07K 14/31 (2006.01)
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195
from bacteria
305
from Micrococcaceae (F)
31
from Staphylococcus (G)
Applicants:
F. HOFFMANN-LA ROCHE AG [CH/CH]; Grenzacherstrasse 124 4070 Basel, CH (AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BE, BF, BG, BH, BJ, BN, BR, BW, BY, BZ, CA, CF, CG, CH, CI, CL, CM, CN, CO, CR, CU, CY, CZ, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, FR, GA, GB, GD, GE, GH, GM, GN, GQ, GR, GT, GW, HN, HR, HU, ID, IE, IL, IN, IR, IS, IT, JO, JP, KE, KG, KH, KM, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LT, LU, LV, LY, MA, MC, MD, ME, MG, MK, ML, MN, MR, MT, MW, MX, MY, MZ, NA, NE, NG, NI, NL, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SI, SK, SL, SM, SN, ST, SV, SY, SZ, TD, TG, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, UZ, VC, VN, ZA, ZM, ZW)
ROCHE DIAGNOSTICS GMBH [DE/DE]; Sandhofer Straße 116 68305 Mannheim, DE (DE)
ROCHE SEQUENCING SOLUTIONS, INC. [US/US]; 4300 Hacienda Drive Pleasonton, California 94588, US (US)
Inventors:
CRAIG, Timothy; US
HARRIS, Corissa; US
AMBROSO, Mark; US
PORTER, Marshall; US
DIPIETRO, Matthew; US
Agent:
HILDEBRANDT, Martin; DE
BURGER, Alexander; DE
GOEHRING, Frank; DE
JENNI, Wolfgang; DE
MISERA, Simon; DE
NASER, Werner; DE
SKOLAUT, Alexander; DE
EISENMANN, Ulrike; DE
MERTES, Maria; DE
HEINEMEYER, Thomas; DE
Priority Data:
62/357,23030.06.2016US
Title (EN) LONG LIFETIME ALPHA-HEMOLYSIN NANOPORES
(FR) NANOPORES D'ALPHA-HÉMOLYSINE À LONGUE DURÉE DE VIE
Abstract:
(EN) Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L135I in the mature alpha-hemolysin amino acid sequence. The α-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.
(FR) L'invention concerne des variantes de l'alpha-hémolysine comportant au moins une substitution d'acide aminé en H35G, E111N, M113A et/ou K147N dans la séquence d'acides aminés de l'alpha-hémolysine de type sauvage mature. Dans certains exemples, la variante peut comporter une substitution en E111S, M113S, T145S, K147S ou L135I dans la séquence d'acides aminés de l'alpha-hémolysine mature. Les variantes de l'α-hémolysine peuvent également comprendre une substitution en H144A et/ou une série de résidus de glycine allant des résidus 127 à 131 de l'alpha-hémolysine de type sauvage mature. L'invention concerne également des ensembles de nanopores comprenant les variantes de l'alpha-hémolysine, lesdits ensembles ayant des nanopores à plus longue durée de vie. L'invention concerne en outre des variantes qui, en plus d'octroyer une plus longue durée de vie, donnent un temps de montée plus court. Les variantes selon l'invention augmentent donc la durée de vie des nanopores et améliorent l'efficacité et la précision des réactions de séquençage d'ADN en utilisant les nanopores comprenant lesdites variantes.
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 Organization (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (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)
Also published as:
CN109415419EP3478706