Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

纳米SiO_2与有机物分子的亲和性和分散性

介绍纳米SiO2的合成和粒子表面的粗糙形貌以及纳米粒子链具有类似高分子链的弹性力学属性,研究纳米SiO2的亲合性,探讨亲合性对纳米SiO2的分散、改性、应用的影响。研究表明,在极性外加剂(如乙二胺、乙二醇、PEG、PDMS、PTHF、PMMA、PBMA等)存在的情况下,纳米SiO2的弹性粒子链和硅烷醇对SiO2的分散和改性具有显著的影响。带有大烷基侧链的高分子链与SiO2表面的亲合能较低,而带有官能团的高分子链与SiO2表面具有较高的亲合性。     

Isolation of the acidic and basic lectins from winged bean seed (Psophocarpus tetragonolobus (L.) DC)

The acidic and basic lectins of the winged bean were isolated from seed extracts by affinity chromatography on lactose-Sepharose 6B and melibiose Bio-Gel P150, respectively. The lectins isolated from six varieties of Psophocarpus tetragonolobus, collected from different regions of south-east Asia, showed no significant differences in their properties; their subunit M_r and amino acid compositions were the same as those of the acidic and basic lectins isolated by non-affinity methods. On isoelectric focusing the affinity purified acidic lectins showed two major protein components (isolectins) while the affinity purified basic lectins had four protein components. The acidic lectins showed a preference for β-D-galactosides and they agglutinated trypsinised human type A, B and O erythrocytes but not trypsinised rabbit erythrocytes. The basic lectins showed a preference for α-D-galactosides and they agglutinated trypsinised rabbit and human type A and B erythrocytes but not trypsinised human type O erythrocytes.     

Thermodynamic state of reinforced interpenetrating polymer networks

The free energies of mixing of two networks in the interpenetrating polymer network based on crosslinked polyurethane and poly(ester acrylate) have been determined by the vapour sorption method. It was established that the constituent networks in the IPN are not miscible. The introduction of fillers of different chemical nature increases the compatibility. The thermodynamic affinity of the fillers to the individual networks and IPN was estimated. It was established that when the free energy of interaction of one or both components of the IPN with the filler is negative, reinforcement leads to the formation of a compatible and equilibrium system. For fillers having no affinity to the polymers, compatibilization is observed, which is connected with slowing down of phase separation in the system in the presence of filler.     

用微机图象法对脉冲MAG焊接熔池进行观察和控制的研究

本文对微机图象法在脉冲MAG焊接中的应用作了探讨,设计和采用了带有CCD摄象机的微机控制系统,对脉冲MAG焊接熔池进行了观察、监控和图象处理。其结果证明了采用微机图象法来实现实时控制是可行的。     

铝元素的添加对锆基合金非晶形成能力影响的电子结构模型

通过构建非晶合金的团簇模型，利用离散变分法从电子层次研究了Al元素对Zr-Cu非晶合金中团簇稳定性的影响。结果表明，Al原子的引入提高了团簇中原子间的亲和力，并且随着Al原子数的增加，亲和力增大，导致新的团簇产生，从而破坏了非晶的团簇结构，Al含量对Zr-Al-Ni非晶合金的稳定性的影响可通过Fermi能级处的态密度大小来反映。     

膜技术在发酵工业中的应用

介绍了膜技术在发酵工业中的应用,主要包括膜分离和膜生物反应器.重点分析了错流过滤应用中的问题.     

Complementarity determining region residues aspartic acid at H55, serine at H95 and tyrosines at H97 and L96 play important roles in the B72.3 antibody-TAG72 antigen interaction

Structural analysis derived from the crystallographic studyof the chimeric B72.3 antibody illustrated some major atomicinteractions between complementarity determining region (CDR)residues. For example, hydrogen bonds are formed between H35/H95,L50/H97, H53/H55 and H96/L96 respectively. These CDR residuesmay play important roles in the B72.3–TAG72 (antibody-antigen)interaction either by direct interaction with the TAG72 antigenor by maintaining a CDR loop conformation through atomic interactionsbetween CDR residues. In order to confirm these assumptions,we altered these CDR residues by site-directed mutagenesis anddetermined binding affinities of these mutant chimeric antibodiesfor the TAG72 antigen in a solid-phase radioimmunoassay. Wefound that H55, H95, H97 and L96 are important CDR residuesfor the B72.3–TAG72 interaction. Single amino acid substitutionsof aspartic acid and serine by alanine at H55 of CDR2 and atH95 of CDR3 respectively and of tyrosine by phenylalanine atH97 and L96 of CDR3, significantly reduced the binding affinityfor the TAG72 antigen by 20-, 8-, 16- and 45-fold respectively.Therefore, this study reveals some of the requirements for maintainingthe integrity of the B72.3 antibody combining sites.     

四种金属离子亲和层析纯化重组戊型肝炎病毒包涵体的效果比较

目的 选择固相金属亲和层析柱的金属离子,以优化分离纯化重组戊型肝炎病毒包涵体的条件。方法 在同一试验条件下用4种金属离子柱分离纯化目的蛋白。结果 包涵体经不同离子柱层析时目的蛋白的收获率、纯度、洗脱条件各异。结论 镍离子柱纯化效果最佳。     

抗rHu-IFN-αMcAb亲和常数的测定

采用ELISA间接法检测抗原－抗体反应系统中游离抗体量的方法，对本实验室制备的5系抗rHu－IFN－αMcAb进行了亲和力测定，获得比较准确的McAb亲和常数。