超大孔聚合物微球PGMA的親水改性及其在分離純化中的應(yīng)用
發(fā)布時間：2018-10-05 13:43
【摘要】：超大孔聚合物PGMA微球具有疏水性,容易與蛋白發(fā)生非特異性吸附使其失去活性,且制備的微球載量較低。筆者針對這一問題進行如下探究:PGMA微球作為基材,在有機溶劑DMSO中通過環(huán)氧基在其表面接枝葡聚糖,然后通過烯丙基活化方法在堿溶液中多次鍍層葡聚糖,使其表面含有大量親水性基團OH,從而達到親水改性的目的。然后通過紅外光譜儀、掃描電鏡和原子力顯微鏡對改性前后微球的化學組成和形貌進行表征;紅外表征表明葡聚糖接枝到微球上,掃描電鏡和原子力顯微鏡表明改性后微球形貌未發(fā)生改變;非特異性吸附結(jié)果表明,改性前后,其非特異性吸附由原來的60%左右變?yōu)?%左右,與親水性的瓊脂糖微球的非特異性吸附相近,接觸角從153°降低到約為0°,親水性良好。用親水改性的PGMA微球通過環(huán)氧氯丙烷環(huán)氧活化連接耐堿ProteinA親和配基,制備成親和層析介質(zhì),并對其制備條件進行優(yōu)化,對其載量和性能進行表征。掃描電鏡和壓力-流速曲線測試表明改性前后其流通性能變化不大�？疾炝谁h(huán)氧值、配基密度及載量之間的關(guān)系,結(jié)果表明配基密度隨著環(huán)氧值的增大而增大、載量隨著配基密度的增大先增大后減小,在配基密度為4.52 mg/mL時,其動態(tài)載量達到最大值,為16.1 mg/mL�？疾炱湓诓煌Ａ魰r間下介質(zhì)的動態(tài)載量,結(jié)果顯示,隨著保留時間從5 min縮短到1 min,介質(zhì)的動態(tài)載量僅僅下降13%,表明其在快速分離純化上有一定的潛力。用親水改性的PGMA微球連接DEAE陰離子交換配基,制備成陰離子交換介質(zhì),并對其改性前后的形貌進行表征。結(jié)果表明,改性前后微球形貌未改變。并考察了孔徑、配基密度對其動態(tài)載量的影響。結(jié)果表明,隨著孔徑的增大,載量變小,而隨著配基密度的增加,介質(zhì)的載量先增大后減小。還考察其在不同保留時間下介質(zhì)的動態(tài)載量,結(jié)果顯示,隨著保留時間從5 min縮短到1 min,介質(zhì)的動態(tài)載量僅僅下降18%,表明其在快速分離純化上有一定的潛力。
[Abstract]:The macroporous polymer PGMA microspheres are hydrophobic and easily adsorbed with protein to make them lose their activity and the load of the prepared microspheres is low. In order to solve this problem, the author studied that the microspheres were grafted with dextran on the surface by epoxy group in organic solvent DMSO, and then coated with dextran in alkaline solution by allyl activation method. The hydrophilic modification can be achieved by making the surface containing a large number of hydrophilic groups OH,. Then the chemical composition and morphology of the microspheres before and after modification were characterized by infrared spectrometer, scanning electron microscope and atomic force microscope, and the results showed that dextran was grafted onto the microspheres. Scanning electron microscopy and atomic force microscope showed that the morphology of the modified microspheres did not change, and the nonspecific adsorption of the modified microspheres changed from about 60% to about 2% before and after the modification. The contact angle was reduced from 153 擄to about 0 擄, and the hydrophilicity was good. The hydrophilic modified PGMA microspheres were used to connect the alkali-resistant ProteinA affinity ligand with epichlorohydrin epoxy-activated. The hydrophilic and chromatographic media were prepared. The preparation conditions were optimized and the loading and properties of the ligands were characterized. Scanning electron microscopy and pressure-flow curve test showed that the fluidity of the modified material changed little before and after modification. The relationships among epoxide value, ligand density and loading capacity were investigated. The results showed that the ligand density increased with the increase of epoxy value, and the loading increased first and then decreased with the increase of ligand density. The ligand density was 4.52 mg/mL. Its dynamic load reached a maximum of 16.1 mg/mL.. The dynamic loading of the medium at different retention times was investigated. The results showed that the dynamic load of the medium decreased only by 13 with the retention time shortened from 5 min to 1 min, which indicated that it had a certain potential for rapid separation and purification. DEAE anion exchange ligands were connected with hydrophilic PGMA microspheres. Anion exchange media was prepared and its morphology was characterized before and after modification. The results showed that the morphology of the microspheres remained unchanged before and after modification. The effects of pore size and ligand density on the dynamic loading were investigated. The results show that the loading capacity decreases with the increase of pore size, and then decreases with the increase of ligand density. The dynamic load of the medium at different retention times was also investigated. The results showed that the dynamic load of the medium decreased only by 18% with the shortening of the retention time from 5 min to 1 min, indicating that it has a certain potential for rapid separation and purification.
【學位授予單位】：河北科技師范學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O631;O652.6
本文編號：2253614
[Abstract]:The macroporous polymer PGMA microspheres are hydrophobic and easily adsorbed with protein to make them lose their activity and the load of the prepared microspheres is low. In order to solve this problem, the author studied that the microspheres were grafted with dextran on the surface by epoxy group in organic solvent DMSO, and then coated with dextran in alkaline solution by allyl activation method. The hydrophilic modification can be achieved by making the surface containing a large number of hydrophilic groups OH,. Then the chemical composition and morphology of the microspheres before and after modification were characterized by infrared spectrometer, scanning electron microscope and atomic force microscope, and the results showed that dextran was grafted onto the microspheres. Scanning electron microscopy and atomic force microscope showed that the morphology of the modified microspheres did not change, and the nonspecific adsorption of the modified microspheres changed from about 60% to about 2% before and after the modification. The contact angle was reduced from 153 擄to about 0 擄, and the hydrophilicity was good. The hydrophilic modified PGMA microspheres were used to connect the alkali-resistant ProteinA affinity ligand with epichlorohydrin epoxy-activated. The hydrophilic and chromatographic media were prepared. The preparation conditions were optimized and the loading and properties of the ligands were characterized. Scanning electron microscopy and pressure-flow curve test showed that the fluidity of the modified material changed little before and after modification. The relationships among epoxide value, ligand density and loading capacity were investigated. The results showed that the ligand density increased with the increase of epoxy value, and the loading increased first and then decreased with the increase of ligand density. The ligand density was 4.52 mg/mL. Its dynamic load reached a maximum of 16.1 mg/mL.. The dynamic loading of the medium at different retention times was investigated. The results showed that the dynamic load of the medium decreased only by 13 with the retention time shortened from 5 min to 1 min, which indicated that it had a certain potential for rapid separation and purification. DEAE anion exchange ligands were connected with hydrophilic PGMA microspheres. Anion exchange media was prepared and its morphology was characterized before and after modification. The results showed that the morphology of the microspheres remained unchanged before and after modification. The effects of pore size and ligand density on the dynamic loading were investigated. The results show that the loading capacity decreases with the increase of pore size, and then decreases with the increase of ligand density. The dynamic load of the medium at different retention times was also investigated. The results showed that the dynamic load of the medium decreased only by 18% with the shortening of the retention time from 5 min to 1 min, indicating that it has a certain potential for rapid separation and purification.
【學位授予單位】：河北科技師范學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O631;O652.6
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