Directed evolution of high-affinity antibody mimics using mRNA display

We constructed a library of >10(12) unique, covalently coupled mRNA-protein molecules by randomizing three exposed loops of an immunoglobulin-like protein, the tenth fibronectin type III domain (10Fn3). The antibody mimics that bound TNF-alpha were isolated from the library using mRNA display. Ten rounds of selection produced 10Fn3 variants that bound TNF-alpha with dissociation constants (K(d)) between 1 and 24 nM. After affinity maturation, the lowest K(d) measured was 20 pM. Selected antibody mimics were shown to capture TNF-alpha when immobilized in a protein microarray. 10Fn3-based scaffold libraries and mRNA-display allow the isolation of high-affinity, specific antigen binding proteins; potential applications of such binding proteins include diagnostic protein microarrays and protein therapeutics.     

Fatty acids of Neem oil

Proceedings - Mathematical Sciences - The work of Khudaet al. relating to the isolation of new fatty acids from Neem oil has been examined and their experiments repeated as far as possible. No...     

Hydroxy-rhodium(I) catalyzed regioselective Michael addition of cyclic enones

Hydroxy-rhodium catalyzed dimerization of various cyclic enones to the corresponding α-enone adducts is developed. The key feature of this method is that the base-sensitive, highly substituted enones also undergo dimerization and the resultant products are obtained in moderate to good yields.     

Polyhydroxybutyrate/Chitosan 3D Scaffolds Promote In Vitro and In Vivo Chondrogenesis

Applied Biochemistry and Biotechnology - The articular cartilage is an avascular and aneural tissue and its injuries result mostly in osteoarthritic changes and formation of fibrous tissue. Efforts...     

A flexible enantioselective synthesis of (+)-centrolobine and 5-epi-diospongin-A using asymmetric transfer hydrogenation/tandem Grubbs cross-metathesis/oxy-Michael reaction as key steps

An efficient enantioselective synthesis of (+)-centrolobine and 5-epi-diospongin-A was achieved by the use of asymmetric transfer hydrogenation (ATH)/tandem Grubbs cross-metathesis/oxy-Michael reaction. Furthermore, this strategy allows for diastereodivergent access to every representative member of the family.     

Synthesis and Characterization of Donor–π‐Acceptor‐Based Porphyrin Sensitizers: Potential Application of Dye‐Sensitized Solar Cells

New porphyrin sensitizers based on donor–π‐acceptor (D‐π‐A) approach have been designed, synthesized, characterized by various spectroscopic techniques and their photovoltaic properties explored. N,N′‐Diphenylamine acts as donor, the porphyrin is the π‐spacer, and either carboxylic acid or cyanoacryclic acid acts as acceptor. All compounds were characterized by using ¹H NMR spectroscopy, ESI‐MS, UV–visible emission spectroscopies as well as electrochemical methods. The presence of aromatic groups between porphyrin π‐plane and acceptor group push the absorption of both Soret and Q‐bands of porphyrin towards the red region. The electrochemical properties suggests that LUMO of these sensitizers above the TiO₂ conduction band. Finally, the device was fabricated using liquid redox electrolyte (I^?/I₃^?) and its efficiency was compared with that of a leading sensitizer.     

Seasonal- and age-dependent changes of the structure and chemical composition of the spherites in the midgut gland of the harvestmen Gyas annulatus (Opiliones)

Spherites—round laminated membrane bound structures—are located in the cytoplasm of all cell types of the midgut gland in the harvestmen Gyas annulatus. The spherites consist of an organic matrix composed of glycoproteins and proteoglycans whose sugar portion are carboxylated glycosaminoglycans. Different elements are embedded in this matrix, and their presence varies during the life cycle. All spherites in juveniles are composed of alternating concentrically arranged electron dense and electron lucent layers of organic matrix material before overwintering (December). At that time, spherites contain calcium, phosphorus and silicon. Calcium and phosphorus are located in their electron dense layers, and silicon spotwise, mainly peripheral. Material seems to be lost during overwintering of Gyas as electron lucent ‘empty’ layers appear in spherites in March. The ‘lost’ material could be used as energy supply and/or to provide molecules for synthesis processes during non-nourishing overwintering. Spherites do not contain calcium and phosphorus in July and October. These elements seem to have important biological relevance in the formation and hardening of the cuticle during the moultings in spring. In contrast to calcium and phosphorus, silicon is still stored in spherites in July and October but in decreasing concentration, therefore it could be involved in metabolic processes in adult Gyas. In the period from July to the end of their adult life in late autumn, an increasing number of spherites disintegrate and their remnant organic matrix material progressively aggregates with other cellular waste material in one or more huge vacuoles of glandular cells.     

Selection of horseradish peroxidase variants with enhanced enantioselectivity by yeast surface display

We report a method for in vitro selection of catalytically active enzymes from large libraries of variants displayed on the surface of the yeast S. cerevisiae. Two libraries, each containing approximately 2 x 10(6) variants of horseradish peroxidase (HRP), were constructed; one involved error-prone PCR that sampled mutations throughout the coding sequence, whereas the other involved complete combinatorial enumeration of five positions near the active site to non-cysteine residues. The enzyme variants displayed on the yeast surface were allowed to modify it with a fluorescently labeled substrate. A combination of positive and negative selection applied to the active-site-directed library resulted in variants with up to an 8-fold altered enantioselectivity, including its reversal, toward L/D-tyrosinol. In contrast, the library constructed by using error-prone PCR yielded no HRP variants with a significantly improved enantioselectivity.