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溫敏性納米水凝膠顆粒的合成及對多肽的識別研究

發(fā)布時間:2019-02-12 09:16
【摘要】:溫敏性納米水凝膠是一類對溫度敏感的高分子聚合物納米水凝膠材料,可以溶于水或者水溶液,具有最低臨界溶解溫度。近年來,由N-異丙基丙烯酰胺(NIPAm)聚合而成的溫敏性水凝膠引起人們的普遍關(guān)注。 本研究以NIPAm為結(jié)構(gòu)框架和溫敏性單體、丙烯酸(AAc)為親電作用單體、N-叔丁基丙烯酰胺(tBAm)為疏水作用單體、N,N’-亞甲基雙丙烯酰胺(BIS)為交聯(lián)劑、十二烷基硫酸鈉(SDS)為表面活性劑、N,N,N’,N’-四甲基乙二胺(Temed)為助聯(lián)劑、過硫酸銨(APS)為引發(fā)劑、在水溶液中制備出一系列溫敏性納米水凝膠顆粒。經(jīng)掃描電鏡(SEM)和動態(tài)光散射儀(DLS)表征,,粒徑均勻。實驗考察了合成配方中SDS和AAc用量對顆粒粒徑的影響。SDS用量從0~50mg變化,粒徑變化為460~70nm。AAc從5%~40%(mol%),粒徑變化為74~613nm,且電位絕對值增加,體系趨于穩(wěn)定。納米水凝膠顆粒在低于15℃時粒徑較大并保持均一穩(wěn)定狀態(tài),隨著溫度上升,尤其當溫度上升至20-30℃之間時,顆粒粒徑開始急速縮小。 將所制備的納米水凝膠顆粒用于組氨酸標簽多肽(NH2-LEVLFQGPHHHHHH-COOH/HOOC-LEVLFQGPHHHHHH-NH2)的識別,經(jīng)高效液相色譜分析,納米水凝膠顆粒粒徑越小,對目標多肽吸附率越高;AAc (負電荷單體)摩爾濃度對吸附率影響較大,采用NIPAm/tBAm/AAc/BIS(43/40/15/2, mol%)配方所得納米水凝膠顆粒對目標多肽吸附率最高,達78%;納米水凝膠顆粒對目標多肽吸附率隨著吸附溶液pH值下降而上升。 將所制備的納米水凝膠顆粒用于蜂毒肽(NH2-GLY-ILE-GLY-ALA-VAL-LEU-LYS-VAL-LEU-THR-THR-GLY-LEU-PRO-ALA-LEU-ILE-SER-TRP-ILE-LYS-ARG-LYS-ARG-GLN-GLN-COOH)的識別,經(jīng)高效液相色譜、聚丙烯酰胺凝膠電泳、紫外-可見分光光度計、細胞體外觀察,隨著納米水凝膠粒徑減小,其對蜂毒肽吸附作用逐漸升高;疏水單體tBAm的配比是影響吸附率的最重要因素,隨著此單體配比的上升,其對蜂毒肽吸附率增大;AAc的配比對其吸附率影響不大。通過調(diào)節(jié)溫度,利用納米水凝膠的溫敏性特征和其在不同粒徑下對蜂毒肽吸附率的區(qū)別,可以控制納米水凝膠顆粒對蜂毒肽吸附-脫附的過程。
[Abstract]:Temperature-sensitive nano-hydrogels are a kind of thermo-sensitive polymer nano-hydrogels which can dissolve in water or aqueous solution and have the lowest critical dissolution temperature. In recent years, thermosensitive hydrogels polymerized by N-isopropyl acrylamide (NIPAm) have attracted much attention. In this study, NIPAm was used as the structure frame and temperature sensitive monomer, acrylic acid (AAc) as electrophilic monomer, N- tert-Ding Ji acrylamide (tBAm) as hydrophobic monomer, and NNT-methylene bisacrylamide (BIS) as crosslinking agent. A series of thermo-sensitive nanogel particles were prepared in aqueous solution with sodium dodecyl sulfate (SDS) as surfactant, NNX (N (N) (n () (n) (n () The particle size was uniform by scanning electron microscope (SEM) and dynamic light scattering (DLS). The effects of the amount of SDS and AAc on the particle size were investigated. The content of SDS changed from 0~50mg to 540% (mol%) for 460~70nm.AAc, and the particle size changed from 74 ~ 613 nm. The absolute value of potential increased and the system tended to be stable. The size of nano-hydrogel particles is large and stable at lower than 15 鈩

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