Genetic engineering to enhance the selectivity of protein separations

The ability to recover and purify natural and recombinant proteins, and the costs of doing so remain a major task in introducing the potential products of biotechnology. The bases for separation range from specific binding onto tailored reagents to solubility and partitioning behavior governed by a mixed bag of size, charge, and hydrophobicity. In most cases, a combination of methods is used in sequence, and improvements in the selectivity at an early stage can enhance the effectiveness of subsequent (and usually more costly) steps. Genetic engineering provides a means of improving the selectivity within the context of existing separation methods. By this strategy, improvements in selectivity are sought by bestowing a distinctive property on the protein of interest. The primary sequence of amino acids is altered, such that the protein can be selectively removed from other components of the multicomponent mixture in which such products are commonly found. In this article, the range of these “distinctive properties” and their pairing with various separation methods will be reviewed. Specific examples from our work, in which a distinctive charge is provided via a polypeptide “purification” fusion tail, will be discussed. Separation methods we have used with these fusion proteins are precipitation, two-phase aqueous extraction, reversed micellar extraction, and ion exchange using both resins and membranes.     

Fibers and 3D mesh scaffolds from biodegradable starch-based blends: production and characterization

The aim of this work is the production of fibers from biodegradable polymers to obtain 3D scaffolds for tissue engineering of hard tissues. The scaffolds required for this highly demanding application need to have, as well as the biological and mechanical characteristics, a high degree of porosity with suitable dimensions for cell seeding and proliferation. Furthermore, the open cell porosity should have adequate interconnectivity for a continuous flow of nutrients and outflow of cell metabolic residues as well as to allow cell growth into confluent layers. Blends of corn starch, a natural biodegradable polymer, with other synthetic polymers (poly(ethylene vinyl alcohol), poly(epsilon-caprolactone), poly(lactic acid)) were selected for this work because of their good balance of properties, namely biocompatibility, processability and mechanical properties. Melt spinning was used to produce fibers from all the blends and 3D meshes from one of the starch-poly(lactic acid) blends. The experimental characterization included the evaluation of the tensile mechanical properties and thermal properties of the fibers and the compression stiffness, porosity and degradation behavior of the 3D meshes. Light microscopy picture of 3D meshes.     

新工科项目“地方(行业)高校化学类专业应对产业变化的升级改造研究与实践”的实施

This "emerging engineering education (3E)" project "Upgrade and Practice of Chemistry-related Majors of Local and/or Specialized University Responding to the Industry Changes" was supported by the Ministry of Education. To aim at the issue of how to upgrade the chemistry-related majors of a local and/or specialized university responding to industry changes, an investigation on local regional economy and industry changes was carried out, the capability requirements of 3E for talents were analyzed, and the training objectives, graduation requirements as well as the new teaching models were also studied for the major to meet the future requirements. The practical experience for 3E construction of chemistry-related majors by selected universities was suggested.     

Linking Chiral Clusters with Molybdate Building Blocks: From Homochiral Helical Supramolecular Arrays to Coordination Helices

The synthesis of four new oxo‐centered Fe clusters ( 1 a – c , 2 ) of the form [Fe^III₃(μ₃‐O)(CH₂=CHCOO)₆] with acrylate as the bridging ligand gives rise to potentially intrinsically chiral oxo‐centered {M₃} trimers that show a tendency to spontaneously resolve upon crystallization. For instance, 1 a , [Fe^III₃(μ₃‐O)(CH₂=CHCOO)₆‐(H₂O)₃]⁺, crystallizes in the chiral space group P3₁ as a chloride salt. Crystallization of 1 b , [Fe₃(μ₃‐O)(C₂H₃CO₂)₆(H₂O)₃]NO₃?4.5H₂O, from aqueous solution followed by recrystallization from acetonitrile also gives rise to spontaneous resolution to yield the homochiral salt [Fe₃(μ₃‐O)(C₂H₃CO₂)₆‐(H₂O)₃]NO₃?CH₃CN of 1 c (space group P2₁2₁2₁). Furthermore, the reaction of 1 a with hexamolybdate in acetonitrile gives the helical coordination polymer {[(Fe₃(μ₃‐O)L₆(H₂O))(MoO₄)‐(Fe₃(μ₃‐O)L₆(H₂O)₂)]?2CH₃CN?H₂O}_∞ 2 (L: H₂C?CHCOO), which crystallizes in the space group P2₁. The nature of the ligand geometry allows the formation of atropisomers in both the discrete ( 1 a – c ) and linked {Fe₃} clusters ( 2 ), which is described along with a magnetic analysis of 1 a and 2 .     

低热-高压法制备PLGA多孔支架及其体外降解研究

采用低热-高压法制备了聚(dl-丙交酯／乙交酯)75／25(PLGA75／25)组织工程多孔支架。该方法避免了使用有机溶剂，支架的孔隙率在90％以上，孔径大小分布均匀。多孔支架经过酒精处理后，支架表面产生许多微小的凹陷；用藻酸钙改性处理后，支架形态保持良好。两种处理都使支架的压缩强度有所增大，亲水性增强。虽然孔隙率高的支架降解速率稍慢，但其体外降解规律基本一致：特性粘数争力学强度衰减快，而质量损失较慢，降解6周后，支架的质量损失仅为3％左右；体外降解3周后，支架的形态保持良好，可望在细胞移植争组织修复的早期发挥支撑作用。     

Thermally produced biodegradable scaffolds for cartilage tissue engineering

A novel process was developed to fabricate biodegradable polymer scaffolds for tissue engineering applications, without using organic solvents. Solvent residues in scaffolds fabricated by processes involving organic solvents may damage cells transplanted onto the scaffolds or tissue near the transplantation site. Poly(L-lactic acid) (PLLA) powder and NaCl particles in a mold were compressed and subsequently heated at 180 degrees C (near the PLLA melting temperature) for 3 min. The heat treatment caused the polymer particles to fuse and form a continuous matrix containing entrapped NaCl particles. After dissolving the NaCl salts, which served as a porogen, porous biodegradable PLLA scaffolds were formed. The scaffold porosity and pore size were controlled by adjusting the NaCl/PLLA weight ratio and the NaCl particle size. The characteristics of the scaffolds were compared to those of scaffolds fabricated using a conventional solvent casting/particulate leaching (SC/PL) process, in terms of pore structure, pore-size distribution, and mechanical properties. A scanning electron microscopic examination showed highly interconnected and open pore structures in the scaffolds fabricated using the thermal process, whereas the SC/PL process yielded scaffolds with less interconnected and closed pore structures. Mercury intrusion porosimetry revealed that the thermally produced scaffolds had a much more uniform distribution of pore sizes than the SC/PL process. The utility of the thermally produced scaffolds was demonstrated by engineering cartilaginous tissues in vivo. In summary, the thermal process developed in this study yields tissue-engineering scaffolds with more favorable characteristics, with respect to, freedom from organic solvents, pore structure, and size distribution than the SC/PL process. Moreover, the thermal process could also be used to fabricate scaffolds from polymers that are insoluble in organic solvents, such as poly(glycolic acid). Cartilage tissue regenerated from thermally produced PLLA scaffold.     

Novel photopolymerizable biodegradable triblock polymers for tissue engineering scaffolds: synthesis and characterization

For use in micro-patterned scaffolds in tissue engineering, novel diacrylated triblock macromers (PLA-b-PCL-b-PLA, PGA-b-PCL-b-PGA and PCL-b-PEO-b-PCL) were synthesized and characterized by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance spectroscopy (NMR) and gel permeation chromatography (GPC). All diacrylated polymers were designed as triblock copolymers and involved biodegradable blocks of relatively non-polar epsilon-caprolactone (CL) and polar monomers such as glycolide (GA), lactide (LA) or ethylene oxide (EO). All triblock polymers were prepared in molecular weights of a few kilo daltons via the anionic ring-opening polymerization (ROP) of the corresponding lactide, glycolide or caprolactone using stannous octoate [Sn(Oct)(2)] as catalyst. The polymers had low polydispersity indices, ranging from 1.23 to 1.56. Biodegradable polymeric networks were prepared with conversions of 72-84% via photopolymerization of the triblock diacrylated polymers with 2,2-dimethoxy-2-phenylacetophenone (DMPA) as photoinitiator. PLA-b-PCL-b-PLA copolymers crumbled easily and were not suitable for micro-patterning. PGA-b-PCL-b-PGA copolymers had higher water contact angles than PCL-b-PEO-b-PCL and were also cytocompatible with Fibroblasts 3T3.     

化学专业生物化学课程教学内容和教学方法研究

本文是笔者在长期教学实践的基础上，对于化学专业生物化学课程教学内容和教学方法，研究的经济总结。其相关成果曾获陕西师范大学优秀教材一等奖及陕西省教学成果二等奖。     

生物催化在绿色化学和新药开发中的应用

近年来生物技术领域有了突破性进展，如: 公共基因数据库(GenBank)和蛋白质数据库(PDB)中序列的指数增长，高效基因克隆和表达平台的建立，可有效改进生物催化剂专一性、选择性和稳定性的酶定向进化技术的应用。这些进展使生物催化在化学合成中日趋重要[1]。本文综述了生物催化在如下领域的成功应用：在药物生产中用于开发经济的化学酶法合成工艺，在绿色化学领域中最大程度的减少废物的产生和危险试剂的应用，在天然化学领域中对天然产物进行修饰以发现具有更好生物活性的新药物。     

材料化学实验课程体系的改革与建设

结合材料化学实验课程自身的特点及材料化学实验教学的经验，系统阐述我院材料化学实验改革的思路，通过深化材料化学实验教学改革，提高任课教师的学术水平，更新教学内容，优化教学模式，推进教材的系统性建设，实现材料化学实验教学质量的提高，激发学生的学习热情与兴趣，进一步培养学生的创新能力。