本文關(guān)鍵詞：自行加工的非對(duì)稱流場(chǎng)流分離儀器對(duì)蛋白質(zhì)分離的應(yīng)用研究　出處：《中國(guó)藥學(xué)雜志》2016年16期 　論文類型：期刊論文

  更多相關(guān)文章： 場(chǎng)流分離 非對(duì)稱流場(chǎng)流 蛋白質(zhì) 分離儀器

【摘要】：目的設(shè)計(jì)并加工一套針對(duì)生物大分子分離的非對(duì)稱流場(chǎng)流分離(asymmetric flow field-flow fractionation,As Fl FFF或AF4)儀器,對(duì)儀器性能進(jìn)行初步研究,為下一步實(shí)際樣品的分離提供依據(jù)和參考經(jīng)驗(yàn)。方法為了解決樣品聚焦(focus)問(wèn)題,自行研究了一種新的聚焦方式。以標(biāo)準(zhǔn)牛血清白蛋白和乙醇脫氫酶作為混合樣品,對(duì)儀器性能進(jìn)行驗(yàn)證。并通過(guò)改變進(jìn)樣/聚焦時(shí)間、橫向流動(dòng)速度(cross-in flow)和橫向流出速度(cross-out flow),考察各因素對(duì)分離效果(分離度R表示)的影響。結(jié)果混合蛋白的分離度分別隨聚焦時(shí)間的減少和橫向流動(dòng)速度的增加而增加,而橫向流出速度的影響并不顯著。結(jié)論該非對(duì)稱流場(chǎng)流分離儀器已基本實(shí)現(xiàn)對(duì)混合蛋白的分離,預(yù)計(jì)可用于更為復(fù)雜的樣品分離。
[Abstract]:Objective to design and process a set of asymmetric flow field-flow fractionation for the separation of biological macromolecules. In order to solve the problem of focusing on samples, the performance of as F1 FFF or AF4 instrument is studied preliminarily, which provides the basis and reference experience for the separation of practical samples in the next step. A new focusing method was studied. The standard bovine serum albumin (BSA) and ethanol dehydrogenase (EDH) were used as mixed samples to verify the performance of the instrument, and the injection / focus time was changed. The lateral flow velocity was cross-flow) and the lateral outflow velocity was cross-out. The results showed that the separation degree of mixed protein increased with the decrease of focusing time and the increase of lateral flow velocity. Conclusion the asymmetric flow separation instrument has basically achieved the separation of mixed proteins, which can be used for more complex sample separation.
【作者單位】： 北京大學(xué)藥學(xué)院;中國(guó)農(nóng)業(yè)大學(xué);
【分類號(hào)】：TH79
【正文快照】： 生物大分子的分離表征一直是困擾研究者的重要問(wèn)題,現(xiàn)在常用的分子生物學(xué)手段均無(wú)法實(shí)現(xiàn)溫和、快速、高效的分離,存在樣品前處理復(fù)雜,分離時(shí)間長(zhǎng),尺寸分辨率低,對(duì)高分子量顆粒無(wú)法分離等缺點(diǎn)[1-2]。本實(shí)驗(yàn)開(kāi)發(fā)的非對(duì)稱流場(chǎng)流分離儀器能分離和表征幾納米至數(shù)十微米的顆粒,可彌

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