静电力驱动蛋白质在疏水蛋白表面的吸附

疏水蛋白是丝状真菌产生的一种外泌蛋白质, 它们可以在不同表面形成双亲性蛋白膜. 疏水蛋白也是一种优良的蛋白质固定化基质, 然而蛋白质在疏水蛋白表面吸附的驱动机制却是未知的. 本文系统研究了不同pH和离子浓度下蛋白质在疏水蛋白表面的吸附. 首先, 用石英晶体微天平技术研究了不同pH和离子浓度下, Ⅰ型疏水蛋白HGFI和Ⅱ型疏水蛋白HFBI在聚苯乙烯表面的吸附. 结果发现, pH和离子强度对HGFI在聚苯乙烯表面的吸附影响较大, 对HFBI的吸附影响与HGFI相比则较小; HGFI在聚苯乙烯表面主要形成的是弹性膜, 而HFBI在聚苯乙烯表面主要形成的是刚性膜. 随后又研究了不同pH和离子浓度下牛血清白蛋白(BSA)和亲和素(Avidin)在HGFI和HFB上吸附, 结果表明, pH和离子强度对BSA和Avidin在HGFI和HFB上吸附有显著影响, 说明BSA和Avidin在两种疏水蛋白上吸附的主要驱动力为静电力. 本文研究结果为实现疏水蛋白表面可控地固定蛋白质提供了理论指导.     

聚苯乙烯-g-聚氧乙烯接枝共聚物表面阻抗蛋白质吸附的研究

利用放射性碘标记技术研究了血浆蛋白质-缓冲溶液体系在聚苯乙烯-g-聚氧乙烯接枝共聚物表面的等温吸附和吸附动力学。材料表面蛋白质等温吸附量或平衡吸附量随表面PEO含量增加而下降;吸附量最低值小于0.25μg,cm^-2;同时探讨了材料表面”梳状“结构对材料表面PEO侧链阻抗蛋白质性能的影响。     

聚苯乙烯—g—聚氧乙烯接枝共聚物表面阻抗蛋白质吸附的研究

利用放射性磺标记技术研究了血浆蛋白质－缓冲溶液体系在聚苯乙烯－ｇ－聚氧乙烯接枝共聚物表面的等温吸附和吸附动力学，材料表面蛋白质等温吸附量或平衡吸附量随表面ＰＥＯ含量增加而下降，吸附量最低值小于０．２５μｇ．ｃｍ＾－２；同时探讨了材料表面“梳状”结构对材料表面ＰＥＯ侧链阻抗蛋白质性能的影响。     

用于免疫分析的聚苯乙烯微孔板功能化研究

为满足固相时间分辨荧光免疫分析研究需要,将聚苯乙烯微孔板内表面固相功能化,研制一种在聚苯乙烯微孔板内表面形成耐受强酸、强碱、有机溶剂的尼龙6膜层,并在膜层表面烷基化得到活性基团与己二酸二酰肼进行"手臂"连接,用双功能团戊二醛活化.检测结果表明,经修饰的聚苯乙烯微孔板化学连接蛋白质结合率比物理吸附高出5～10倍,在-4℃环境中存放6个月活性降低小于15%.灵敏度和稳定性显著提高.     

含1,2,3-三氮唑官能团嵌段共聚物的制备及其蛋白质的吸附

通过"点击化学"反应合成了含1,2,3-三氮唑官能团的单体(VBTM),再采用可逆加成-断裂链转移(RAFT)活性自由基聚合法,制备了含有1,2,3-三氮唑官能团的聚苯乙烯(PS-b-PVBTM)两嵌段共聚物。将PS-b-PVBTM旋涂于金片表面成膜,形成以聚苯乙烯链段为核、PVBTM链段为冠的胶束结构;此薄膜于90℃退火24 h后,球状结构转变为表面富集PS的环状结构。采用表面等离子体谐振仪(SPR)原位检测蛋白质在PS-b-PVBTM上的吸附动力学。结果表明,当缓冲液为中性时,聚合物薄膜对牛血清蛋白(BSA)和肌红蛋白(Myoglobin)没有吸附,而对等电点为11的溶菌酶(Lysozyme)则有吸附;经退火处理后的聚合物薄膜对Lysozyme的吸附量减少了0.26 ng/mm2;当缓冲液为酸性时,该聚合物薄膜对BSA产生吸附,且吸附量随缓冲液pH值的降低而增加。PS-b-PVBTM对以上3种蛋白质的吸附为静电吸附。     

高度交联聚苯乙烯吸附剂的结构与吸附性能研究

在通过Ｆｒｉｅｄｅｌ—Ｃｒａｆｔｓ后交联反应制备高度交联聚苯乙烯吸附剂的过程中，向反应体系中加入非极性芳香化合物甲苯，当加入少量甲苯时，吸附剂的比表面积、孔容等变小，但其吸附能力却明显提高；当加入甲苯超过某一量时，其吸附能力又随加入甲苯的量的增加而降低，这说明孔结构和骨架结构共同影响着高度交联聚苯乙烯吸附剂的吸附性能．     

超高交联聚苯乙烯对非水体系中苯酚的吸附热力学研究

采用“一锅法”以氯化锌为催化剂合成了超高交联聚苯乙烯。测定了超高交联聚苯乙烯对环己烷中苯酚的吸附等温曲线，利用热力学函数关系计算出了吸附焓、吸附自由能和吸附熵，比较了超高交联聚苯乙烯对苯酚和苯甲醚吸附性能的差别，推测吸附过程的主要作用为氢键作用。以小分子化合物为模型，通过分子间相互作用能的计算，论证了树脂是通过其羰基氧原子与苯酚的羟基氢原子之间的氢键作用而吸附苯酚的。     

PNIPAAm改性表面对蛋白质吸附的调控及其应用

根据不同领域的需要,控制蛋白质在材料表面的吸附是一个具有重要应用价值的课题。聚（N-异丙基丙烯酰胺）（PNIPAAm）改性表面能够响应外界温度变化从而改变其表面性质,这一特点为调控蛋白质的吸附提供了可能。近年来,研究者们应用多种表征方法考察了不同温度下蛋白质在PNIPAAm改性表面的吸附,并试图从分子水平上深入理解其吸附机制及影响因素。本文综述了近年来应用PNIPAAm改性表面对蛋白质吸附的研究及其最新进展。发现当PNIPAAm层厚度处于一定范围内时,PNIPAAm改性表面表现出对蛋白质吸附的温度敏感性,并可以利用这一性质将其应用于蛋白质纯化及分离和生物传感器等领域。而当PNIPAAm层厚度超过一个临界值时,PNIPAAm改性表面表现出良好的阻抗血浆蛋白质的性质,使其有望在血液相容性表面领域得到应用。最后,就PNIPAAm改性表面调控蛋白质吸附的未来发展方向简要地进行了展望。     

硫辛酸在三种不同树脂上的吸附热力学和动力学研究

通过静态吸附实验,研究了大孔交联聚苯乙烯树脂(XAD-4)、氧修饰超高交联聚苯乙烯树脂(NDA-100)和胺基修饰超高交联聚苯乙烯树脂(ND-90)对乙醇．水溶液中硫辛酸的吸附热力学及动力学特性,结果表明：硫辛酸在XAD-4树脂上是单层吸附,符合Langmuir等温吸附方程,吸附过程符合准一级动力学吸附方程。硫辛酸在NDA-100和ND-90树脂上的吸附也符合Langmuir等温吸附方程,但并不只是单层吸附,同时兼有毛细管凝聚和微孔填充作用,吸附过程可分为大孔和中孔区的吸附以及微孔区的吸附两个阶段,两个阶段都符合准一级动力学吸附方程。     

表面活性剂在低能固体表面上的吸附 Ⅲ: 聚苯乙烯胶乳粒子对阴离子表面活性剂的吸附

在无乳化剂的条件下合成了粒径均匀的聚苯乙烯胶乳, 发展了应用表面张力计测定吸附等温线的连续平衡法, 得到不同盐浓度下聚苯乙烯胶乳对十二烷基硫酸钠和十二烷基苯磺酸钠的吸附等温线, 它们属于Giles分类的L2 型或L1 型, 采有两阶段吸附模式讨论了吸附机理, 吸附层结构及等温线类型变化的规律。     

Surface immobilization of poly(ethylene oxide): Structure and properties

We covalently immobilized poly(ethylene oxide) (PEO) chains onto a fluorinated ethylene propylene copolymer (FEP) surface. On the FEP surface, aldehyde groups were first deposited by plasma polymerization of acetaldehyde or acrolein. Then, amino‐PEO chains were immobilized through Schiff base formation, which was followed by reduction stabilization with sodium cyanoborohydride. The PEO‐grafted polymer surfaces thus prepared were characterized by X‐ray photoelectron spectroscopy (XPS), atomic force microscopy, contact‐angle measurements, and protein adsorption. The dramatic increase in the C O intensity of the high‐resolution XPS C 1s spectrum, together with an overall increase in oxygen content, indicated the successful attachment of PEO chains onto the acetaldehyde plasma surfaces. The amount of grafted PEO chains depended on the superfacial density of the plasma‐generated aldehyde groups. The grafted monoamino‐PEO chains formed a brushlike structure on the polymer surface, whereas the bisamino‐PEO chains predominately adopted a looplike conformation. The PEO surface had a regular morphology with greater roughness than the aldehyde surface underneath. Surface hydrophilicity increased with the grafting of PEO. Also, the bisamino‐PEO‐grafted surface had slightly higher surface hydrophilicity than its monoamino‐PEO counterpart. These PEO coatings reduced fibrinogen adsorption by 43% compared with the substrate FEP surface. © 2000 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 38: 2323–2332, 2000     

Immobilization of protein on cellulose hydrogel

A technique of immobilizing an enzyme/antibody was developed using cellulose hydrogel prepared from an aqueous alkali-urea solvent. Partial oxidation by sodium periodate activated the cellulose gel for introducing aldehyde groups. Proteins were covalently introduced to cellulose gel by a Schiff base formation between the aldehyde and the amino groups of proteins, and stabilized by a reduction of imines. Coloring reactions confirmed the high activity of the immobilized enzymes. The activity of the immobilized enzymes increased with aldehyde content, but the effect leveled off at a low degree of oxidation, at approximately 8.1 of oxidized glucose/100 glucose unit. The amount of immobilized peroxidase calculated from the activity was 8.0 ng/g for an aldehyde content of 0.18 mmol/g: 14.6 ng/g for both 0.46 mmol/g and 1.04 mmol/g. The same method could be applied to the peroxidase antibody. Thus, various active proteins could be immobilized on cellulose gels by mild and facile processing. Owing to high mechanical and chemical stability of cellulose, this technique and resulting materials are potentially useful in biochemical processing and sensing technologies.     

Photoresponsive crown ethers. 10. Metal complexation by light-switched crown ethers immobilized in polymer matrices

The crosslinked polystyrene beads which immobilized two terminal azophenoxide groups of an azobenzene-crown(24-crown-8)-azobenzene unit were synthesized. The polymer beads adsorbed Cs⁺ in the dark while they rapidly released Cs⁺ into the solution under UV-light irradiation, and the photoresponsive complexation occurred reversibly. On the other hand, the crosslinked polystyrene beads which immobilized one terminal azophenoxide group of an azobenzene-crown (24-crown-8) unit scarcely (or only to a smaller extent) exhibited such a photoresponsive behavior. Similarly, the polymer beads which immobilized two terminal azophenoxide groups of an azobenzene-crown (18-crown-6) azobenzene unit reversibly adsorbed and desorbed K⁺ in response to UV-light irradiation. The novel photoresponsive behavior is ascribed to a conformational distortion of the crown rings which induced by the photoisomerization of the azobenzene moiety, although the photoinduced polarity change of the polymer beads is not completely ruled out. The results suggest that the polymer support is useful as a “fixed point” to enforce the conformational changes of immobilized functional molecules.     

Attachment of horseradish peroxidase (HRP) onto the poly(styrene/acrolein) latexes and onto their derivatives with amino groups on the surface; activity of immobilized enzyme

The polystyrene (P(S)), poly(styrene/acrolein) (P(SA)), and polyacrolein (P(A)) latexes, with varied fraction of polyacrolein in the surface layer (f _A=0, 0.50, 0.63, 0.84, 1.00), were used for the attachment of horseradish peroxidase. Surfaces of latexes were modified by reaction with ethylenediamine. In this step the aldehyde groups from polyacrolein were blocked and the primary amino groups were introduced. The carbohydrate portion of HRP was oxidized in the reaction leading to formation of aldehyde groups. The adsorption and covalent immobilization of HRP onto the P(S), P(SA), and P(A) latexes and of the oxidized HRP (HRP-OX) onto the modified latex particles, with amino groups on the surface (P(SA)-M and P(A)-M), were investigated. The activities of parent and oxidized HRP were compared with activities of the corresponding enzymes in solution. It has been found that whereas HRP is not suitable for the covalent immobilization on P(SA) latex and loses its activity after adsorption onto P(S) latex, HRP-OX can be adsorbed onto P(S) latex and is readily immobilized covalently onto the ethylenediamine modified P(SA) and P(A) latexes, retaining much of its former enzymatic reactivity.This work was supported by the KBN Grant 2 0624 91 01     

用无乳化剂乳液聚合法制备聚丙烯醛微球

本文应用无乳化剂乳液聚合法合成了粒度均一、具有活泼醛基的聚丙烯醛微球,并对丙烯醛聚合动力学和影响微球直径,活泼醛基含量的因素进行了研究。用合成的微球同不同的氨基酸和午血清白蛋白反应,结果表明:该微球在生理pH条件下同含有伯氨基的化合物具有较高的结合容量。因此,在无乳化剂存在下,采用乳液聚合法制备聚丙烯醛微球是一种合成具有干净表面的聚丙烯醛微球的新方法。     

The Synthesis and Characterization of Polyacrolein through Radical Polymerization

A radical polymerization reaction of acrolein is reported in this article. The free radical initiator which can effectively promote the free radical polymerization of acrolein is screened out. The optimal conditions of the reaction are investigated and the yield could be up to 93.67%, in which the ratio of initiator to monomer is 1:50, monomer concentration is 7.5 mol L^?1, reaction temperature is 50 °C, and the reaction time is 6 h. The structure characterizations of the obtained polymers are performed using hydrogen nuclear magnetic resonance spectroscopy, fouriertransform infrared spectroscopy, and matrix‐assisted laser desorption ionization time of fligh mass spectroscopy. The results show that the structure of the polymer contains fragments generated by decomposition of the initiator, aldehyde groups, and vinyl groups. The reaction mechanism of acrolein polymerization in the presence of free radical initiator is proposed. Thus, a novel method for the preparation of polyacrolein via radical polymerization is provided in this article.     

相转移催化法实现聚苯乙烯的功能化转变

以对羟基苯甲醛的钠盐为亲核取代试剂,采用相转移催化进行了大分子氯甲基聚苯乙烯的亲核取代反应,将其转变为侧链带有功能性基团醛基的聚苯乙烯;考察了反应条件(如有机溶剂的极性、催化剂种类及用量、有机相与水相之比等)对大分子相转移催化反应的影响,并根据相转移催化反应机理与相关的动力学规律进行了分析.结果表明,通过相转移催化亲核取代反应,可将氯甲基聚苯乙烯大分子链上的氯原子转化,制得侧链带有醛基的聚苯乙烯;溶剂的极性越强,负离子的反应活性也越高;催化剂季铵正离子的亲脂性越强,相转移催化反应的速率越快.当催化剂浓度较低时,四丁基溴化铵比十六烷基溴化铵的催化效果好;当催化剂浓度较高时,十六烷基溴化铵比四丁基溴化铵的催化效果好.另外,有机相与水相之比对相转移催化反应有较大的影响.     

Short-term measurements of acrolein in ambient air

Summary A method has been developed for the determination of acrolein in air samples collected by a high-volume aqueous scrubber. The aldehyde is collected as the bisulfite adduct, which is decomposed before determination of acrolein by DNPH (2,4-dinitrophenylhydrazine) derivatization and HPLC. Approximately 95% of the acrolein reacts with DNPH within 3 h at DNPH:HSO₃ molar ratios of up to 10. The method appears promising for short-term air sampling at 8 L min^–1, enabling the achievement of a detection limit of 0.2 g m^–3 for acrolein.     

Graft polymerization from pore wall of three-dimensionally ordered macroporous cross-linked polystyrene via atom transfer radical polymerization

The surface-initiated atom-transfer radical polymerization (ATRP) technique was applied to the graft polymerization of methyl methacrylate (MMA) and N-isopropylacrylamide (NIPAm) from three-dimensionally ordered macroporous cross-linked polystyrene (3DOM CLPS) on which the initiator, halogen atom was immobilized onto the pore wall of 3DOM CLPS by chloromethylation. FT-IR and TG-DWA analyses confirm that the graft polymerization of MMA and NIPAm via ATRP had been taken place at the pore wall of 3DOM CLPS. The initiating efficiency of chloromethyl groups was calculated according to the data of TGA-titration, revealed that the benzyl chloride is not only distributed on the surface of the pore walls but also must be present throughout the inner of the cross-linked polystyrene matrix. SEM analyses show that the grafted layers are smooth and homogeneous, and the ordered structure is well preserved after polymerization. By the adjustment of the graft polymerization time, the thickness of grafted polymer layers can be controlled. The max thickness of grafted PMMA layer is 85 nm and the max thickness of grafted PNIPAm layer is 35 nm.     

纤维素固定化血红蛋白研究

纤维素是一种重要的化工原料,由于来源广泛、价廉以及良好的再生性、反应性和通用性,被广泛用做载体材料．在酶的分离纯化,蛋白质分离纯化,亲合色谱固定相,拆分手性化合物等应用领域,往往采用球形纤维素或膜状纤维素形式．而在水吸附剂,分离回收金属离子,从海水中...