詳細(xì)介紹
IC693CPU360
IC693CPU360
《;歐》
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《:同步》
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目前上普遍采用液體處理機(jī)器人來進(jìn)行高通量結(jié)晶篩選,篩選一個(gè)結(jié)晶條件的蛋白樣品消耗體積為1-5微升。為了降低單次篩選所需的蛋白質(zhì)樣品體積,工業(yè)界普遍采用微型化的策略來降低zui小的液體操縱體積。這當(dāng)中具有代表性的有英國Douglas公司研制的Oryx-nano系列結(jié)晶機(jī)器人、美國ArtRobbins公司研制的Phoenix機(jī)器人、英國TTPLabTech公司研制的Mosquito系列機(jī)器人等。
采用這些篩選機(jī)器人,單個(gè)篩選反應(yīng)的蛋白質(zhì)消耗量可降低到50–200nL,比常規(guī)篩選系統(tǒng)降低了20–100倍,不僅顯著地降低了蛋白質(zhì)結(jié)晶篩選所需的蛋白質(zhì)樣品消耗量,也有效減少了研究成本和時(shí)間,因而已成為部分結(jié)構(gòu)生物學(xué)實(shí)驗(yàn)室的標(biāo)準(zhǔn)配置。盡管如此,對于膜蛋白等一些難以表達(dá)純化的*蛋白質(zhì)樣品,這類儀器的蛋白質(zhì)耗樣量仍然過高,難以滿足結(jié)構(gòu)生物學(xué)研究的需要。
為了解決這一矛盾,浙江大學(xué)研究團(tuán)隊(duì)發(fā)展了一種基于微流控液滴技術(shù)和自動化機(jī)器人技術(shù)的超微量蛋白質(zhì)結(jié)晶和篩選系統(tǒng)。該系統(tǒng)采用順序操作液滴陣列模式構(gòu)建,同時(shí)采用微加工技術(shù),在一個(gè)1平方厘米的微孔陣列區(qū)域集成了676個(gè)獨(dú)立的微孔陣列用于高密度的蛋白質(zhì)結(jié)晶孵育和篩選。
通過研究微觀多相流體的特點(diǎn),結(jié)合系統(tǒng)性裝置優(yōu)化和改進(jìn),將液體處理機(jī)器人對蛋白質(zhì)和沉淀劑樣品的zui小操控體積降低至120皮升,比現(xiàn)有機(jī)器人系統(tǒng)降低了約200-800倍。
該裝置被用于對5種水溶性蛋白和1種膜蛋白的大規(guī)模結(jié)晶條件篩選,得到了與常規(guī)篩選體系接近的篩選結(jié)果。單個(gè)液滴結(jié)晶反應(yīng)器的體積為4納升,包含2納升的蛋白質(zhì)和2納升的沉淀劑。篩選96種沉淀劑條件,每種條件重復(fù)三次,共生成288個(gè)液滴結(jié)晶反應(yīng)器,蛋白質(zhì)溶液的總消耗量僅為579nL,與商品化篩選系統(tǒng)相比,試劑消耗量降低了2-3個(gè)數(shù)量級。
由于蛋白質(zhì)消耗量的顯著降低,該裝置還可以用于快速生成蛋白質(zhì)二維結(jié)晶相圖以及研究微觀體系下蛋白質(zhì)結(jié)晶的特殊規(guī)律,為蛋白質(zhì)結(jié)晶的理論研究提供一種有力的工具。
MINTERTIA UGRMEM-02MAK0E
MISC DTS-2303
MISC MP40LK08
MITSUBISHI 305A141A
MITSUBISHI 56H1CSBN
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MODICON 120-00A2-001
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MODULAR CONTROLS SV4-10-03-115AS
MOOG 164-001E-10-B2-2
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MOOG UNKNOWN
MOOG X303-030A
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MOTION CONTROL 41-04DC0005D0AC007
MOTION CONTROL SYSTEMS 674A-302
MOTOR NO TAG SMALL SILVER
MOTOR POWER T142SR255RH17501E0014
MOTOR POWER CO ESA6L9201
MOVINIR M70105-0001
MOVINOR M7010S0001
MSI MTE-404-500-MA
MTS MPM1141T2-1521
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MTS AUTOMATION MPM1902-1675
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N/A 42301483-80-B
NACHI MFA450FSCRS
NACHI TS4074E28
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NACHI USADED-15-FC31
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NACHI-FUJIKOSHI MFA450FSCBS
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NEC DFSM3020502A
NECKAR-MOTOREN M563-456600700
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NEUGART PL90FA
NIDEK MINERTIA J03M-XE41
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NISSEN VAFP-22-30-4002
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ORIENTAL 4IJ15A-A
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ORIENTAL MOTOR 3IK15GN-AUL
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ORMEC 06-068
ORMEC M52PE52
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ORMEC MAC-B020A/K1A
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OSAI OSS-F6200B
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P.A.U 500185
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