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蛋白質(zhì)純化方法-親和層析親和層析法包括特異性配體親和層析法以及通用性配體親和層析法。比較這兩種親和層析法。特異性配體親和層析法通常將復(fù)雜的生命大分子物質(zhì)作為配體，它的結(jié)合力比較大河吸附選擇性比較強(qiáng)。而通用性配體親和層析法通常將簡單的小分子物質(zhì)作為配體，因此，它具有低廉的成本，吸附容量比較高，通過對吸附和脫附條件的改善能夠使得層析的分辨率提高，浙江化學(xué)分析蛋白純化系統(tǒng)。原理親和層析的原理類似于酶-底物，浙江化學(xué)分析蛋白純化系統(tǒng)，***-受體與抗原-抗體等特異性反應(yīng)的機(jī)理。有一定的親和力在每對反應(yīng)物之間。與在酶與底物的反應(yīng)中，只有特異的底物才可以結(jié)合一定的酶，使復(fù)合物產(chǎn)生相同。在親和層析中，浙江化學(xué)分析蛋白純化系統(tǒng)，只有特異的配體才可以和一定的生命大分子有親和力，并且使復(fù)合物產(chǎn)生。親和層析配體呈固相，而酶與底物的反應(yīng)底物呈液相是它們的不同點(diǎn)。事實(shí)上，親和層析是在含有活化基團(tuán)的基質(zhì)M上以共價鍵的方式固化具有識別能力的配體L，將固相載體制作成功而固化后的配體束縛特異物質(zhì)的能力依然保持。所以當(dāng)在小層析柱裝入固相載體，使得欲分離的樣品在該柱經(jīng)過。此時，樣品中對配體有親和力的物質(zhì)S就能夠通過結(jié)構(gòu)互補(bǔ)效應(yīng)、范德瓦爾力與靜電引力等作用在固相載體上吸附著。浙江化學(xué)分析蛋白純化系統(tǒng)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="浙江化學(xué)分析蛋白純化系統(tǒng),蛋白純化系統(tǒng)" src="http://tyunfile.71360.com/UpLoadFile/szyss/2020/1/637138171674741306/6371381716747413062985954.jpg"></div><p style="text-indent:25px">蛋白質(zhì)分離純化步驟（二） 2.4.2 纖維素柱層析法該法是利用蛋白質(zhì)的酸堿性質(zhì)作為分離的基礎(chǔ)。離子交換纖維素（cellulose ion-exchanger）是人工合成的纖維素衍生物，它具有松散的親水性網(wǎng)狀結(jié)構(gòu)，有較大的表面積，使蛋白質(zhì)大分子可以自由通過。因此常用于蛋白質(zhì)的分離。（1）羧甲基纖維素（CM-纖維素）在纖維素顆粒上帶有羧甲基基團(tuán)。在中性pH條件下，羧甲基上的質(zhì)子可解離下來（圖2-27a），而溶液中帶正電荷的蛋白質(zhì)分子可與纖維素顆粒上的羧甲基負(fù)電荷結(jié)合。可交換的基團(tuán)帶正電，因此是一種陽離子交換劑。蛋白質(zhì)與離子交換纖維素之間結(jié)合能力的大小取決于彼此間相反電荷基團(tuán)之間靜電吸引。 (2）二乙氨基乙基纖維素（DEAE-纖維素）在中性pH條件下，它含有帶正電荷的基團(tuán)，可與溶液中的帶負(fù)電荷的蛋白質(zhì)結(jié)合，可交換的基團(tuán)帶負(fù)電荷，因此是一種陰離子交換劑。當(dāng)某一蛋白質(zhì)混合溶液通過裝有DEAE-纖維素的層析柱時，帶正電荷的蛋白質(zhì)不能結(jié)合而隨著洗脫液的流動先被洗脫下來。帶負(fù)電荷的蛋白質(zhì)將被結(jié)合到柱上。結(jié)合力取決于彼此相反電荷基團(tuán)間的靜電吸引。然后選用一定pH和離子強(qiáng)度的緩沖液進(jìn)行洗脫，改變蛋白質(zhì)分子所帶的靜電荷，依次從層析柱流出達(dá)到相互分離的目的。浙江化學(xué)分析蛋白純化系統(tǒng)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="浙江化學(xué)分析蛋白純化系統(tǒng),蛋白純化系統(tǒng)" src="http://tyunfile.71360.com/UpLoadFile/szyss/2020/1/637138172066720957/6371381720667209577791021.jpg"></div><p style="text-indent:25px">三、蛋白質(zhì)分子量的測定 3.3 沉降法沉降法又稱超速離心法。蛋白質(zhì)溶液在受到強(qiáng)大的離心力作用時，蛋白質(zhì)分子趨于下沉，沉降速度與蛋白質(zhì)分子的大小、密度和分子形狀有關(guān)，也與溶劑的密度和粘度有關(guān)。蛋白質(zhì)顆粒在離心場中的沉降速度用每單位時間內(nèi)顆粒下沉的距離來表示。在離心場中，蛋白質(zhì)分子所受到的凈離心力（離心力減去浮力）與溶劑的摩擦力平衡時，每單位離心場強(qiáng)度的沉降速度稱為沉降系數(shù)（Sedimentation Coefficient）。國際上采用Svedberg單位作為沉降系數(shù)的單位，用S表示，以紀(jì)念超速離心法的創(chuàng)始人，瑞典***的蛋白質(zhì)化學(xué)家T.Svedberg。一個Svedberg單位（或直接稱一個S）為1×10—13s。蛋白質(zhì)的沉降系數(shù)大約在1×10—13～200×10—13s范圍內(nèi)，即1S～200S。</p><p style="text-indent:25px">蛋白質(zhì)分離純化步驟（二） 2.4 樣品的進(jìn)一步分離純化用等電點(diǎn)沉淀法、鹽析法所得到的蛋白質(zhì)一般含有其他蛋白質(zhì)雜質(zhì)，須進(jìn)一步分離提純才能得到有一定純度的樣品。常用的純化方法有：凝膠過濾層析、離子交換纖維素層析、親和層析等等。有時還需要這幾種方法聯(lián)合使用才能得到較高純度的蛋白質(zhì)樣品。 1. 凝膠過濾層析凝膠過濾層析（gel-filtration chromatography）又稱為分子排阻層析或分子篩層析。它是將葡聚糖凝膠（Sephadex）裝入一個柱子中，制成凝膠柱。這種凝膠顆粒具有網(wǎng)狀結(jié)構(gòu)，不同類型凝膠的網(wǎng)孔大小不同。當(dāng)把蛋白質(zhì)混合樣品加到凝膠柱中時，比凝膠網(wǎng)孔小的蛋白質(zhì)可進(jìn)入網(wǎng)孔內(nèi)，而大于網(wǎng)孔的分子則不能進(jìn)入被排阻在凝膠顆粒之外。當(dāng)用洗脫液洗脫時，被排阻的分子量大的蛋白質(zhì)直接通過凝膠之間的縫隙先被洗脫下來，而比網(wǎng)孔小的蛋白質(zhì)可連續(xù)不斷地進(jìn)入網(wǎng)孔內(nèi)。這樣的小分子不但流經(jīng)的路程長，而且受到來自凝膠內(nèi)部的阻力也很大，所以蛋白質(zhì)越小，從柱子上洗脫下來所需時間越長。由于不同蛋白質(zhì)的分子大小不同，進(jìn)入網(wǎng)孔的程度不同，因此流出的速度不同，洗脫所用體積及時間不同，從而達(dá)到分離的目的。</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="浙江化學(xué)分析蛋白純化系統(tǒng),蛋白純化系統(tǒng)" src="http://tyunfile.71360.com/UpLoadFile/szyss/2020/1/637138171561815352/6371381715618153525073938.jpg"></div><p style="text-indent:25px">三、蛋白質(zhì)分子量的測定蛋白質(zhì)分子量測定的方法很多，目前常用的方法有以下幾種。 3.1 凝膠過濾法凝膠過濾法測定蛋白質(zhì)分子量的原理如“分離純化方法”中所述。一定型號的凝膠顆粒上具有一定大小的孔隙，只允許較小的分子進(jìn)入膠粒，而大于孔隙的分子則不能進(jìn)入膠粒而被排阻在膠粒外面。用洗脫液洗脫時，被排阻的分子量大的蛋白質(zhì)先被洗脫下來，隨后能夠進(jìn)入顆粒的蛋白質(zhì)也按分子量大小而先后被洗脫下來，分子量越小的越后被洗脫下來。由于不同排阻范圍的葡聚糖凝膠有一特定的蛋白質(zhì)分子量范圍，在此范圍內(nèi)，分子量的對數(shù)和洗脫體積之間成線性關(guān)系。因此，用幾種已知分子量的蛋白質(zhì)為標(biāo)準(zhǔn)進(jìn)行層析分析，以每種蛋白質(zhì)的洗脫體積對它們的分子量的對數(shù)作圖，繪制出標(biāo)準(zhǔn)洗脫曲線。未知蛋白質(zhì)在同樣的條件下進(jìn)行層析分析，根據(jù)其所用的洗脫體積，從標(biāo)準(zhǔn)洗脫曲線上可求出此未知蛋白質(zhì)對應(yīng)的分子量來。北京智能蛋白純化系統(tǒng)上門維修</p><p style="text-indent:25px">浙江化學(xué)分析蛋白純化系統(tǒng)</p><p style="text-indent:25px">儀器儀表是用以檢出、測量、觀察、計算各種物理量、物質(zhì)成分、物性參數(shù)等的器具或設(shè)備。真空檢漏儀、壓力表、測長儀、顯微鏡、乘法器等均屬于儀器儀表。近年來，得益于機(jī)械、冶金、石化行業(yè)等儀器儀表服務(wù)領(lǐng)域經(jīng)營狀況的好轉(zhuǎn)，我國儀器儀表制造業(yè)發(fā)展一路向好。隨著互聯(lián)網(wǎng)的逐步發(fā)展，為蛋白純化色譜系統(tǒng)，凝膠凈化色譜系統(tǒng)，制備液相色譜，柱塞泵等產(chǎn)品的傳播提供了一個飛速的平臺。讓儀器儀表行業(yè)從傳統(tǒng)的銷售模式到以互聯(lián)網(wǎng)電子商務(wù)為主的營銷方式的轉(zhuǎn)變，促進(jìn)了儀器儀表行業(yè)與互聯(lián)網(wǎng)的結(jié)合，推動產(chǎn)業(yè)創(chuàng)新發(fā)展。中國儀器儀表行業(yè)目前正處于高速發(fā)展階段，需要與之相適應(yīng)的機(jī)器人與自動化設(shè)備、機(jī)械電子設(shè)備的研發(fā)、生產(chǎn)、銷售及技術(shù)咨詢；光學(xué)檢測設(shè)備、光電產(chǎn)品研發(fā)、制造、銷售；實(shí)驗(yàn)室設(shè)備、儀器儀表的研發(fā)、制造、銷售及提供相關(guān)的技術(shù)咨詢和售后服務(wù)；自營和代理各類商品及技術(shù)的進(jìn)出口業(yè)務(wù)（限定企業(yè)經(jīng)營或禁止進(jìn)出口的商品和技術(shù)除外）。（依法需經(jīng)批準(zhǔn)的項(xiàng)目，經(jīng)相關(guān)部門批準(zhǔn)后方可開展經(jīng)營活動）產(chǎn)品營銷模式相互配合。從加工廣義角度來說，儀器儀表也可具有自動操控、報警、信號傳遞和數(shù)據(jù)處理等功能，例如用于工業(yè)生產(chǎn)過程自動操控中的氣動調(diào)節(jié)儀表，和電動調(diào)節(jié)儀表，以及集散型儀表操控系統(tǒng)也皆屬于儀器儀表。浙江化學(xué)分析蛋白純化系統(tǒng)</p><p 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