以聚乙烯基咪唑为配基的内毒素亲和吸附剂的研究

通过乙烯基咪唑(VI)在硅胶粒子表面的自由基接枝聚合制备了一种以聚乙烯基咪唑为配基的新型内毒素亲和吸附剂. 用FTIR检测样品中咪唑基的特征吸收, 用热重分析法(TGA)测定了PVI的接枝率. 实验发现, PVI在吸附剂中的含量对内毒素的吸附率影响很大. 当PVI的接枝率为2.5％左右时, 吸附剂对内毒素的去除率最大. 在离子强度小于1 mol/L和pH=7的中性条件下, PVI吸附剂对内毒素具有最佳的吸附性能. 该吸附剂具有良好的血液相容性. 内毒素在该亲和吸附剂上的吸附等温线符合Freundlich吸附方程, 其吸附动力学为二级反应.     

计算机模拟内毒素吸附剂吸附机理的研究

应用计算机模拟的方法研究了内毒素吸附剂的吸附机理. 模拟结果显示, 以二甲胺为配体的吸附剂, 当β位存在羟基时, 此羟基可与内毒素分子间形成氢键, 并形成一个八元环的稳定结构. 此时吸附剂与内毒素之间存在静电、 氢键、 疏水相互作用和八元环的协同作用. 同时模拟了羟基位于配体不同位置的吸附剂与内毒素的相互作用. 结果表明, 静电作用为主要的相互作用力, 羟基的位置对吸附剂的吸附能力影响显著.     

以壳聚糖为载体的内毒素吸附剂

内毒素血症(Endotoxemia)可出现于多种疾病过程中,导致器官坏死、不可逆休克和死亡。如何及时并有效地清除患者体内的内毒素,是临床医学面临的一个难题．选用高效吸附剂,籍助血液灌流的方法从血液中直接清除内毒素,受到了人们越来越多的关注。     

血液灌流用内毒素吸附材料研究

内毒素血症是败血症致死率极高的主要原因之一,迅速并有效地清除患者体内的内毒素,是临床医学面临的一个难题．用血液灌流的方法,选用有效的吸附剂,通过体外循环从血液中直接清除内毒素,受到了人们越来越多的关注．国内外研究者已开始研究利用该方法治疗内毒素血症,用活性炭     

吸附剂结构的计算机辅助设计及吸附机理

以琼脂糖凝胶为载体, 二甲胺为配体, 制备了β-OH, γ-OH和β-SH吸附剂. 通过对水相中内毒素的动、静态吸附探索有效的吸附剂结构. 研究结果显示, 吸附剂对内毒素的清除率随配体结构的不同呈现出较大差异, 其中β-OH吸附剂的清除效果最好, 达到90.7%. 运用计算机模拟方法提出了β-OH吸附剂与内毒素作用的模型, 阐明静电力是影响吸附性能的主要因素, 同时氢键的协同和空间位阻效应也不可忽视.     

亲和吸附剂对细菌内毒素吸附性能的研究

制备了以球形纤维素为载体、8种氨基酸和1种聚赖氨酸为配基的吸附剂,对质量浓度为100.0pg/mL的内毒素水溶液进行了吸附研究,绘制了吸附等温线,并初步探讨了吸附机理.结果表明,精氨酸和赖氨酸配基具有良好的吸附能力,在1.5mL100.0pg/mL内毒素溶液中吸附量分别达到182.0和160.0pg/mL;吸附等温线显示,以赖氨酸为配基的吸附剂其吸附量随溶液内毒素浓度增加而线性增加,符合Langmuir吸附方程,吸附能力强,具有一定的临床应用前景.     

尿毒症中分子毒物吸附剂的研究(Ⅲ)——酸性氨基酸修饰交联壳聚糖

以球状壳聚糖为载体,酸性氨基酸为配体,合成了尿毒症中分子毒物吸附剂,并测试了其血液相容性.为了研究吸附剂对尿毒症患者体内多肽蓄积物的吸附性能,选取分子量不同的6种多肽作为体内蓄积多肽的模拟物进行吸附实验.研究表明,吸附剂对多肽模拟物产生一定的吸附作用.在此基础上又对尿毒症患者血清超滤液进行了吸附实验,结果显示,吸附剂对血清中分子级分的清除率为10.3%.     

血液净化高分子吸附材料

简要总结了我们近年来在血液净化高分子吸附材料方面取得的研究成果. 对于有机小分子吸附剂、中分子吸附剂和生物大分子吸附剂的系统研究,不仅取得了大量的基础性研究结果,而且筛选出了性能优良的血液净化吸附剂,已经与血液灌流装置一起开始应用于临床.     

一种新型低密度脂蛋白吸附剂的制备

研究表明,人体血浆中低密度脂蛋白(LDL)的含量过高会引起动脉硬化,进而诱发各种心脑血管疾病^[1].近年来,采用血液净化吸附剂去除LDL受到了广泛的关注^[2～10].目前临床上使用的吸附剂主要有固载LDL抗体的琼脂糖吸附剂及固载肝素或磺化葡聚糖的纤维素吸附剂,它们虽有较好的吸附效果,但由于存在配基昂贵、吸附剂成本高及机械强度差等问题,这些吸附剂的应用受到了很大的限制.本文以具有良好机械强度和血液相容性的聚乙烯醇(PVA)大孔珠状树脂为载体,以廉价易得的吲哚-3-乙酸(IAA)为配基,分别以不同长度的多胺为悬臂,制备了一种新型LDL吸附剂,并初步考察了其对LDL的吸附性能.     

HA型大孔吸附树脂及活性炭对胆红素吸附性能的研究

本文选用HA型大孔吸附剂对患者血中胆红素进行了动态、静态吸附性能实验,考察了其对血液生化的影响。同时,用瑞典GAMBRO肾罐中活性炭进行了比较,实验结果表明,HA吸附剂对直接胆红素的吸附功效与进口活性基本相同,但血液生化指标HA吸附剂优于活性炭。     

以赖氨酸为配基的内毒素吸附剂的研究

内毒素血症可出现于多种疾病中,如大面积烧伤、重症肝炎、肝硬化等疾病,使机体免疫功能严重受损,引起多器官功能衰竭等一系列严重的病理变化,最终导致不可逆休克和死亡,在美国每年都有超过10万人死于此病,临床上尚无有效的治疗方法,因此,及时、有效地清除或破坏患者体内的内毒素,     

ADSORBENTS USED IN THE CLEARANCE OF ENDOTOXIN

A series of modified poly (methyl methacrylate, PMMA) resins were prepared and compared their adsorption abilities to endotoxin. The results showed that adsorbents, which were grafted with tertiary amine and long spacing arms, had the best adsorption capacities and good blood compatibility, It is hopeful to be used as adsorbent in hemoperfusion for clinical clearance of endotoxin. The influence of original concentration of endotoxin on adsorption and the adsorption mechanism were also investigated.     

ADSORBENTS USED IN THE CLEARANCE OF ENDOTOXIN

1.INTRODUCTION[()-227.3()]Endotoxinislipopolysaccharide(LPS)derivedfromthecellmembranesofgram-negativebacteria.Endotoxincancausefebrilereactionsinanimalswithsymptomsofhighfever,vasodilation,diarrheaandfetalshockwheninjectedeveninaverysmallamount[1].Iftheconcentrationofendotoxininpatients?bloodishighenoughitcancauseseveresepsis,whichisamajorcauseofdeathinpatientsandcontinuestohaveahighmortalitydespiteappropriatesurgery,potentantibiotic,andintensivesupportivetherapy[2~5].LipidAisthemaintox…     

Removal of endotoxin from culture supernatant of Bordetella pertussis with aminated poly(gamma-methyl L-glutamate) spherical beads.

Attempts were made to prepare adsorbents having a high affinity for endotoxin in the culture supernatant of Bordetella pertussis. When poly(gamma-methyl L-glutamate) (PMLG) was used as a matrix and amino groups as the ligand, the highest affinity for endotoxin was attained even at a high ionic strength (mu = 0.2-0.4). PMLG beads containing amino groups of about 3.2 meq/g selectively removed endotoxin from the culture supernatant of B. pertussis without affecting the protective antigens. It was demonstrated that 1 ml of the wet adsorbent adsorbed 4.5 mg of endotoxin. The beads of PMLG derivatives, therefore, are considered to be a useful adsorbent for the removal of endotoxin from the pertussis vaccine, affecting neither filamentous hemagglutinin nor pertussis toxin.     

New family of glutathionyl-biomimetic ligands for affinity chromatography of glutathione-recognising enzymes

Three anthraquinone glutathionyl-biomimetic dye ligands, comprising as terminal biomimetic moiety glutathione analogues (glutathionesulfonic acid, S-methyl-glutathione and glutathione) were synthesised and characterised. The biomimetic ligands were immobilised on agarose gel and the affinity adsorbents, together with a nonbiomimetic adsorbent bearing Cibacron Blue 3GA, were studied for their purifying ability for the glutathione-recognising enzymes, NAD+-dependent formaldehyde dehydrogenase (FaDH) from Candida boidinii, NAD(P)+-dependent glutathione reductase from S. cerevisiae (GSHR) and recombinant maize glutathione S-transferase I (GSTI). All biomimetic adsorbents showed higher purifying ability for the target enzymes compared to the nonbiomimetic adsorbent, thus demonstrating their superior effectiveness as affinity chromatography materials. In particular, the affinity adsorbent comprising as terminal biomimetic moiety glutathionesulfonic acid (BM1), exhibited the highest purifying ability for FaDH and GSTI, whereas, the affinity adsorbent comprising as terminal biomimetic moiety methyl-glutathione (BM2) exhibited the highest purifying ability for GSHR. The BM1 adsorbent was integrated in a facile two-step purification procedure for FaDH. The purified enzyme showed a specific activity equal to 79 U/mg and a single band after sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. Molecular modelling was employed to visualise the binding of BM1 with FaDH, indicating favourable positioning of the key structural features of the biomimetic dye. The anthraquinone moiety provides the driving force for the correct positioning of the glutathionyl-biomimetic moiety in the binding site. It is located deep in the active site cleft forming many favourable hydrophobic contacts with hydrophobic residues of the enzyme. The positioning of the glutathione-like biomimetic moiety is primarily achieved by the strong ionic interactions with the Zn2+ ion of FaDH and Arg 114, and by the hydrophobic contacts made with Tyr 92 and Met 140. Molecular models were also produced for the binding of BM1 and BM3 (glutathione-substituted) to GSTI. In both cases the biomimetic dye forms multiple hydrophobic interactions with the enzyme through binding to a surface pocket. While the glutathioine moiety of BM3 is predicted to bind in the crystallographically observed way, an alternative, more favourable mode seems to be responsible for the better purification results achieved with BM1.     

Galactosyl-biomimetic dye-ligands for the purification of Dactylium dendroides galactose oxidase

Two anthraquinone galactosyl-biomimetic dye-ligands comprising, as terminal biomimetic moiety, galactose analogues (1-amino-1-deoxy-beta-D-galactose and D(+)-galactosamine) were designed for the enzyme galactose oxidase (GAO), using molecular modelling, synthesized and characterized. The biomimetic ligands were immobilized on agarose beads and the affinity adsorbents, together with a non-biomimetic adsorbent bearing Cibacron Blue 3GA, were studied for their ability to purify GAO from Dactylium dendroides. Both biomimetic adsorbents showed higher purifying ability for GAO compared to the non-biomimetic adsorbent, thus demonstrating their superior effectiveness as affinity chromatography materials. In particular, the affinity adsorbent comprising, as terminal biomimetic moiety, 1-amino-1-deoxy-beta-D-galactose (BM1) exhibited the highest purifying ability for GAO. This affinity adsorbent did not bind galactose dehydrogenase, glucose dehydrogenase, alcohol dehydrogenase, or glucose oxidase. The dissociation constant (K(D)) of the immobilized BM1 ligand with GAO was found to be equal to 45.8 microM, whereas the binding capacity was equal to 709 U per ml adsorbent. Therefore, the BMI adsorbent was integrated in a facile two-step purification procedure for GAO. The purified enzyme showed a specific activity equal to 2038 U/mg, the highest reported so far, approximately 74% overall recovery and a single band after sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis.