Biotechnology and Applied BiochemistryThis is an RSS file. You can use it to subscribe to this data in your favourite RSS reader, such as GoogleReader, or to display this data on your own website or blog. Subscribe to this data using MyMedWorm.Subscribe to this data using GoogleReader.Subscribe to this data using Bloglines.Subscribe to this data using MyYahoo.

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Preparation, characterization and evaluation of biopolymeric gold nanocomposite with antimicrobial activityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This report describes the antimicrobial activity of chitosan capped gold nanoparticles coupled with ampicillin (C-AuNp-Amp). C-AuNp-Amp was synthesized using wet precipitation method and characterized using Fourier transform infrared spectroscopy (FT-IR) and atomic force microscopy (AFM) techniques. The optimal level of ampicillin concentration that couples with C-AuNp nanocomposite was determined by using UV-Vis spectroscopy. The agar well diffusion method was used to evaluate the antimicrobial activity and the broth dilution assay was used to determine the minimum inhibitory concentration (MIC). The size of the ellipsoid...
Source: Biotechnology and Applied Biochemistry - November 20, 2009 Category: Biochemistry Authors: M Chamundeeswari, S S. Liji Sobhana, J P. Jacob, M Ganesh Kumar, M Pandima Devi, T P. Sastry, A B. Mandal Source Type: journals

Prokaryotic expression, purification and characterization of a novel pro-apoptosis protein hPNAS-4Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study represented an important step to investigate the characterization for the new pro-apoptosis protein hPNAS-4, which aids to discover new drug targets for development of effective therapeutic approaches to cancer in future. (Source: Biotechnology and Applied Biochemistry)
Source: Biotechnology and Applied Biochemistry - November 13, 2009 Category: Biochemistry Authors: H Deng, Q Jiang, S Liang, F Yan, S Hou, Z Qian, J Li, Y Wen, J Yang, Y Wei Source Type: journals

A novel harvesting method for cultured cells using iron cross-linked alginate films as culture substratesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The present study was conducted to assess the efficiency of a novel cell harvesting method involving dissolution of the culture substrate composed of ferric ion-cross linked alginate (Fe-alginate). Cell harvesting is an essential step for recovery of cultured adherent cells, but conventional methods such as trypsinization or scraping cause considerable damage to the cells. We therefore devised an original method for harvesting cultured cells using Fe-alginate films as a culture substrate, and then retrieving the cells by disintegration of the alginate gel. Fe-alginate was easily dissolved under physiological conditions by ...
Source: Biotechnology and Applied Biochemistry - November 11, 2009 Category: Biochemistry Authors: I Machida-Sano, Y Matsuda, H Namiki Source Type: journals

Features of bacterial growth and polysaccharide production of Streptococcus pneumoniae serotype 14Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The effect of several cultivation conditions on the kinetics of bacterial growth and polysaccharide production of S. pneumoniae serotype 14 was studied. The presence in the supernatant of serotype specific capsular polysaccharide (CPS) during growth was followed by size exclusion HPLC and in parallel confirmed by a specific latex reagent. Agitation level did not affect the production behaviour, whereas pH maintenance above 6 strongly enhanced both growth and CPS production throughout the cultivation period in flasks. Production of high molecular weight polysaccharide was found to be largest between 5 to 6 hr cultivation, a...
Source: Biotechnology and Applied Biochemistry - November 5, 2009 Category: Biochemistry Authors: H Massaldi, M Inés Bessio, N Suárez, E Texeira, S Rossi, F Ferreira Source Type: journals

Effect of IPP5, a novel inhibitor of PP1, on apoptosis and the underlying mechanisms involvedEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Genes encoding apoptosis-inducing proteins are postulated to be candidate tumor suppressors. The identification of such proteins may benefit the early diagnosis and therapy of tumors. In the current study, we characterized the function of a novel human bone marrow stromal cell (BMSC)-derived protein (IPP5) by large scale random sequencing of a human BMSC cDNA library. Human IPP5 cDNA encodes a protein of 116 amino acid residues, which shares high homology with human inhibitor-1 of protein phosphatase 1 (PPI-1). The effect of IPP5 on apoptosis and the underlying molecular mechanisms were investigated by overexpression of IP...
Source: Biotechnology and Applied Biochemistry - October 29, 2009 Category: Biochemistry Authors: Q Zeng, Y Huang, L Zeng, X Lan, Y Huang, S He, H Zhang Source Type: journals

Development of cationic colloidal silica coated magnetic nanospheres for highly selective and rapid enrichment of plasma membrane fractions for proteomics analysisEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, an improved PM isolation technique involving coating intact cells with synthesized cationic silica coated magnetic nanospheres was developed and applied to the proteomic analysis of the PM from human erythroleukemia K562 cells. The Western blotting characterization and the protein identification of the prepared PM indicated that PM enrichment method using the prepared magnetic nanospheres was a fast and inexpensive strategy with high specificity. Our results demonstrate the potential of these cationic silica coated magnetic nanospheres for high-throughput identification of PM proteins from cells. (Source: Bi...
Source: Biotechnology and Applied Biochemistry - October 27, 2009 Category: Biochemistry Authors: X Li, X Jia, C Xie, Y Lin, R Cao, Q He, P Chen, X Wang, S Liang Source Type: journals

In vitro small intestinal epithelial cell growth on a nanocomposite polycaprolactone scaffoldEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In conclusion, as a further step in investigating small intestinal tissue engineering the nanocomposite employed in this study may prove to be a useful alternative to PLGA in the future. (Source: Biotechnology and Applied Biochemistry)
Source: Biotechnology and Applied Biochemistry - October 27, 2009 Category: Biochemistry Authors: A Gupta, D Vara, G Punshon, K Sales, M Winslet, A M Seifalian Source Type: journals

A one-step exclusion-binding procedure for purification of functional heavy-chain and mammalian-type gamma-globulins from camelid seraEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A new approach has recently been proposed for the purification of "mammalian-type" immunoglobulin G (IgG), consisting of exclusion-binding. The technique uses a gel ("Melon gel", PIERCE) that binds to all plasma proteins but not to IgGs, thus allowing IgGs to be recovered in the flow-through fraction. Here, the technique was applied to camelid IgGs, which are known to be constituted not only of classic mammalian-type IgGs (IgG1) but also of IgGs containing only heavy chains (HC-IgGs). Both mammalian type and HC-IgGs can be purified in the flow-through fraction of dromedary plasma samples with less than 8.5% contamination, ...
Source: Biotechnology and Applied Biochemistry - October 13, 2009 Category: Biochemistry Authors: M R Blanc, A Anouassi, M Ahmed Abed, G Tsikis, S Canepa, V Labas, M Belghazi, G Bruneau Source Type: journals

Efficient expression and secretion of recombinant human growth hormone in the methylotropic yeast Pichia pastoris: Potential applications for other proteinsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Abstract A simple high yielding process for production of recombinant human growth hormone (rhGH) in Pichia pastoris system is described. The approach, adopted addition of surfactants during fermentation along with methanol induction. Pichia integrant harboring multiple copy, non-codon optimized hGH, showed poor expression in complex and defined medium. Inclusion of surfactants Tween 20 or Tween 80 during induction, enhanced the expression levels significantly in shake flask studies. Combination of 2 liter basal salt medium (BSM) and Tween 20 in bioreactor culminated in 3x104 folds elevated expression of the protein (~500...
Source: Biotechnology and Applied Biochemistry - October 7, 2009 Category: Biochemistry Authors: A Apte-Deshpande, S Rewanwar, P Kotwal, V Raiker, S Padmanabhan Source Type: journals

Synthesis of model morpholine derivatives with biological activities by laccase-catalyzed reactionsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The efficient enzyme-catalyzed reaction of morpholines as model structures for bioactive compounds with para-dihydroxylated aromatic systems was carried out using the oxidoreductase laccase and atmospheric oxygen to produce eight novel morpholine-substituted aromatics. The laccase of Myceliophthora thermophila was used for cross-linking morpholines containing primary or secondary amino groups with para-dihydroxylated laccase substrates. We demonstrate that not only primary amino groups but also secondary amino groups are able to couple with para-dihydroxylated aromatic systems in laccase-catalyzed reactions. The resulting ...
Source: Biotechnology and Applied Biochemistry - September 29, 2009 Category: Biochemistry Authors: V Hahn, A Mikolasch, K Wende, H Bartrow, U Lindequist, F Schauer Source Type: journals

Molecular cloning, characterization of two tropinone reductases in Anisodus acutangulus and enhancement of tropane alkaloids production in AaTRI-transformed hairy rootsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Two tropinone reductases constitute a branching point in the biosynthesis of tropane alkaloids. Here we report for the first time the cloning and characterization of two full-length cDNAs encoding AaTRI (GenBank Accession No. EU424321) and AaTRII (GenBank Accession No. EU424322) from Solanaceous plant Anisodus acutangulus by rapid amplification of cDNA ends (RACE). Sequence comparison results indicated AaTRI and AaTRII had high homology with other tropinone reductases from Hyoscyamus niger, Datura stramonium etc, but AaTRI and AaTRII only showed identity of 60.8%. Phylogenetic tree analysis results showed that AaTRI and Aa...
Source: Biotechnology and Applied Biochemistry - September 13, 2009 Category: Biochemistry Authors: G Kai, L Li, Y Jiang, X Yan, Y Zhang, X Lu, P Liao, J Chen Source Type: journals

Cell surface display of the active mannanase in Yarrowia lipolytica with a novel surface display systemEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A novel surface display system was constructed using the cell-wall anchor protein Flo1p from Saccharomydes cerevisiae, the mannanase (man1) from Bacillus subtilis was fused with the C-terminal of Flo1p and the 6×His-tag was inserted between Flo1p and man1. The fusion proteins were confirmed to be displayed on the cell surface of Yarrowia lipolytica successfully by immunofluorescence. Then, the surface-displayed mannanase was characterized. The optimum catalytic conditions for the recombinant mannanase was 55℃at pH 6.0, and it exhibited high stability against pH variation. The highest activity of the recombina...
Source: Biotechnology and Applied Biochemistry - August 31, 2009 Category: Biochemistry Authors: X Yang, Z Jiang, H Song, S Jiang, C Madzak, L Ma Source Type: journals

Development of Unsteady-state model for a biological system in miniaturized BioreactorsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study the special shake flasks, so called ventilation flasks find equipped with oxygen sensors and then an unsteady state gas-transfer model for shake flasks was developed and experimentally investigated for a wide range of gas transfer resistances (kplug). For the validation of our unsteady state model to simulate the gas transfer in a biological system in the ventilation flasks, a strain of Corenobacteriu glutamicum DM 1730 was used as a model organism. For further easy processing the resulting total mass-transfer resistance (kplug) is described as a function of the mass flow through the sterile plug (OTRplug) by...
Source: Biotechnology and Applied Biochemistry - August 16, 2009 Category: Biochemistry Authors: G Amoabediny, Y Ziaie-Shirkolaee, J Büchs Source Type: journals

Design of a filter train for precipitate removal in monoclonal antibody downstream processingEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Protein A chromatography has become widely established for the preparative purification of monoclonal antibodies. Low pH elution from Protein A columns followed by neutralization can often lead to precipitation of impurities in the product stream leading to a visually turbid solution. Pre-treatment of the cell culture harvest stream with increased surface area of a depth filter was found to reduce the magnitude of this problem through exploitation of the adsorptive properties of harvest depth filters. However, this was not a complete solution. Clarification of this turbid product stream prior to the polishing chromatograph...
Source: Biotechnology and Applied Biochemistry - August 4, 2009 Category: Biochemistry Authors: S Kandula, S Babu, M Jin, A A. Shukla Source Type: journals

Lysozyme retention on hydrophobic chromatography predicts resin performance at large scaleEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, the lysozyme retention value (LRV), on the vendor Certificate of Analysis (C of A), was predictive of the performance of a hydrophobic interaction (HIC) resin in a complex manufacturing step. This processing step is used to reduce host cell protein impurities (PI) while maintaining a desired glycoform profile of recombinant Thyroid Stimulating Hormone (TSH). A correlation was found between a first moment analysis of the HIC elution peaks and resin LRV. About 91% of the observed variation was accounted for by resin LRV and LRV of 55 and 50 were significantly different. The acceptable LRV range, to maximize gl...
Source: Biotechnology and Applied Biochemistry - August 3, 2009 Category: Biochemistry Authors: F Riske, M Smith, C Zhang, K White Source Type: journals

High-level expression and large-scale preparation of soluble HBx antigen from Escherichia coliEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The hepatitis B virus (HBV) X protein (HBx) is a multifunctional regulator of cellular signal transduction and transcription pathways in host-infected cells. Evidence suggests that HBx has a critical role in the pathogenesis of hepatocellular carcinoma (HCC). However, the lack of efficient large-scale preparation methods for soluble HBx has hindered studies on the structure and function of HBx. Here, a new pMAL-c2x protein fusion and purification system was used for high-level expression of soluble HBx fusion protein. The high-purity fusion protein was obtained via amylose resin and Q-Sepharase chromatography. The untagged...
Source: Biotechnology and Applied Biochemistry - July 15, 2009 Category: Biochemistry Authors: D Liu, L Zou, W Li, L Wang, Y Wu Source Type: journals

The effects of crosslink density and chemistry on the calcification potential of diamine-extended glutaraldehyde-fixed bioprosthetic heart-valve materialsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Despite indications that glutaraldehyde (GA) crosslinked tissues remain prone to long-term degradation and calcification, it is still the reagent of choice in the fixation of bioprosthetic heart valves. We have previously shown that increased GA concentrations and diamine extension of crosslinks with lysine incorporation lead to a mitigated in vivo calcification, mainly of porcine aortic wall tissue. The current study was performed to assess the correlation between the crosslink-density of all three commonly used tissue types (porcine aortic wall (PW), porcine aortic leaflet (PL), and bovine pericardium (BP)) after GA�...
Source: Biotechnology and Applied Biochemistry - June 29, 2009 Category: Biochemistry Authors: D Bezuidenhout, A Oosthuysen, P Human, C Weissenstein, P Zilla Source Type: journals

Generation of tumor necrosis factor alpha-specific affibody molecules capable of blocking receptor binding in vitroEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Affibody molecules specific for human TNFα were selected by phage-display technology from a library based on the 58-residue protein A-derived Z domain. TNFα is a proinflammatory cytokine involved in several inflammatory diseases, and to this day, four TNFα-blocking protein pharmaceuticals have been approved for clinical use. The phage selection generated 18 unique cystein-free affibody sequences of which 12 were chosen, after sequence cluster analysis, for characterization as proteins. Biosensor binding studies of the 12 E. coli-produced and IMAC-purified affibody molecules revealed three variants that...
Source: Biotechnology and Applied Biochemistry - June 22, 2009 Category: Biochemistry Authors: A Jonsson, H Wållberg, N Herne, S Ståhl, F Y Frejd Source Type: journals

Analysis of phospholipids, sterols, and fatty acids in Taxus chinensis var. mairei cells in response to shear stressEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
To obtain a better understanding of responsive mechanism of plant cells in response to hydrodynamic mechanical stress, lipid profiling of suspension cultured Taxus chinensis var. mairei cells under shear stress was carried out using LC-MS/MS and GC-TOF-MS. T. chinensis var. mairei cells cultured in Couette type shear reactor responded with an increase of cell membrane permeability compared with control cells, which indicated adaptation to shear stress altered membrane lipid composition. The main changes of lipid profiles in the shear stress induced cells were the following: a) total phospholipids content decreased, especia...
Source: Biotechnology and Applied Biochemistry - June 16, 2009 Category: Biochemistry Authors: P Han, J Zhou, Y Yuan Source Type: journals

The N-terminal modification increases the stability of the recombinant human endostatin in vitroEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, zinc-binding and zinc-free endostar were compared to further characterize the biochemical and structural properties. Thermal-induced denaturation was determined by monitoring changes in the fluorescence emission spectra. The result indicated that zinc binding significantly increased the transition temperature of endostar and contributed to a reversible change in protein conformation after recooling. Proteolysis assays demonstrated that the modified protein binding with zinc ions can stabilize the N-terminus as well as the C-terminus of endostar when treated with trypsin, chymotrypsin, carboxypeptidase A and ...
Source: Biotechnology and Applied Biochemistry - June 15, 2009 Category: Biochemistry Authors: L Jiang, C Zou, X Yuan, W Luo, Y Wen, Y Chen Source Type: journals

Engineering and characterization of a bispecific HER2{Multiplication sign}EGFR-binding affibody moleculeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The human epidermal growth factor receptor-2 (HER2) and the epidermal growth factor receptor (EGFR) are overexpressed in various forms of cancer, and the co-expression of both HER2 and EGFR has been reported in a number of studies. The simultaneous targeting of HER2 and EGFR has been discussed as a strategy to potentially increase the efficiency and selectivity in molecular imaging and therapy of certain cancers. In an effort to generate a molecule capable of bispecific targeting of HER2 and EGFR, a gene fragment encoding a divalent HER2-binding affibody molecule was genetically fused in frame to a divalent EGFR-binding af...
Source: Biotechnology and Applied Biochemistry - June 3, 2009 Category: Biochemistry Authors: M Friedman, S Lindström, L Ekerljung, H Andersson-Svahn, J Carlsson, H Brismar, L Gedda, F Y. Frejd, S Ståhl Source Type: journals

Use of design of experiments (DOE) for development of refold technology for progenipoietin-1, a recombinant human cytokine fusion protein from E. coli inclusion bodiesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Optimization of refolding conditions for progenipoietin was performed. The molecule has five disulfide bonds and hence, is a challenge to refold. Variables studied included pH, dithiothreitol (DTT) concentration, cystine concentration, urea concentration, protein concentration, dissolution hold time, and oxygen availability. In view of the complexity of the reaction with respect to the number of parameters that can impact the refold efficiency, some variables were examined via single parameter studies while others were looked at via Design of experiments (DOE). The DOE approach allowed us to look at the effect of these var...
Source: Biotechnology and Applied Biochemistry - May 13, 2009 Category: Biochemistry Authors: D M. Boyle, J J. Buckley, G V. Johnson, A Rathore, M E. Gustafson Source Type: journals

Pifithrin-alpha decreases the radioprotective efficacy of Podophyllum hexandrum fraction (REC-2006) in HepG2 cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Inhibition of p53 by pifithrin-α promotes the p53-mediated apoptosis and protects against doxorubicin-induced apoptosis. The present study was carried out to evaluate the effect of pifithrin-α on radioprotective potential of Podophyllum hexandrum fraction (REC-2006) in HepG2 (p53++) cell line. REC-2006 (10-5μg/ml) treatment 2h before irradiation (10Gy) rendered 80±3% protection in HepG2 cells, whereas, pifithrin-α debilitated the radioprotective potential of REC-2006. REC-2006 increased the expression of Hsp70, HSF-1 and Bcl2 in irradiated HepG2 cells, whereas, pifithrin-&#x...
Source: Biotechnology and Applied Biochemistry - May 1, 2009 Category: Biochemistry Authors: P Kumar Singh, R Kumar, A Sharma, R Arora, S Kumar Jain, R Kumar Sharma Source Type: journals

Over-production of human interleukin-2 in recombinant E. coli BL21 high cell density culture by determination and optimization of necessary amino acids using a simple stoichiometric modelEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In order to increase the productivity of human interleukin-2, a stoichiometric model has been used to determine the most needed amino acids and a precise value of their amounts to be added to the culture during expression of human interleukin-2 (as a model protein) by recombinant Escherichia coli BL21 [pET21a-hil2]. Experiments have been performed to investigate the effect of chosen amino acids and their interactions on expression of human interleukin-2. Glutamine, the mixture of leucine, aspartic acid, glycine and the mixture of leucine, glutamine, aspartic acid, were the most effective interaction in expression of interl...
Source: Biotechnology and Applied Biochemistry - April 2, 2009 Category: Biochemistry Authors: S Yegane Sarkandy, A Farnoud, S Shojaosadati, R Khalilzadeh, M Sadeghizadeh, B Ranjbar, V Babaeipour Source Type: journals

Construction and experimental application of a highly efficient temperature-selection T-vectorEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
T-A cloning is the most commonly used molecular cloning technique in which T-vector is the core tool. T-vectors commonly used now are blue-white positive selection vectors based on mechanism of α-complement of β-galactose of E. coli. Disadvantages of these traditional T-vectors in application include insufficient positive rate and the existence of false-positive clones, which, especially the latter, makes screening of transformant clones via colony PCR necessary. In the present study, a novel T-vector, based on the strategy of inhibiting the growth of negative transformants by means of a lethal gene and innov...
Source: Biotechnology and Applied Biochemistry - March 30, 2009 Category: Biochemistry Authors: Y Ma, A Li, Y Wang, M Xie, J Li, S Zhang, Q Wang Source Type: journals

CXCL10 gene therapy efficiently inhibited the growth of cervical carcinoma based on the antiangiogenic and antiviral activityEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Epidemiological studies have demonstrated that high-risk human papillomavirus (HPV) is involved in causing cervical carcinoma. The HPV oncoproteins E6 and E7 immortalize human keratinocytes is mostly resulted from inactivation of tumor suppressor proteins p53 and pRB, which also play an important role in regulating the expression of pro- and antiangiogenic factors. The present study was conducted to determine whether IFN--inducible protein 10 (IP-10)/CXC chemokine ligand 10(CXCL10), one of the potent antiangiogenic chemokines, can inhibit the growth of cervical cancer. Plasmid DNA encoding CXCL10 was encapsulated with cati...
Source: Biotechnology and Applied Biochemistry - March 3, 2009 Category: Biochemistry Authors: L Yang, C Ping, S Luo, J Li, K Liu, H Hu, Y Wei Source Type: journals

Application of the SILAC (stable isotope labeling with amino acids in cell culture) technique in quantitative comparisons for tissue proteome expressionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In conclusion, the Leu-d3-based quantitative technique can be effectively expanded to compare the expression levels for tissue proteome at different states, which allows to identify new candidate biomarkers for tumors. (Source: Biotechnology and Applied Biochemistry)
Source: Biotechnology and Applied Biochemistry - February 26, 2009 Category: Biochemistry Authors: Y Xu, S Liang, G Shen, X Xu, Q Liu, Z Xu, F Gong, M Tang, Y Wei Source Type: journals

Stabilization of naked and condensed plasmid DNA against degradation induced by ultrasounds and high shear vorticesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Micron-sized aggregates of a 6050 base pair plasmid obtained by the addition of 1.5-3.0 mM CaCl2 and 20% v/v t-butanol or 0.3-1.0% v/v aluminium phosphate gel were subjected to degradation induced by sonication or vorticular flows. Dynamic light scattering revealed that the plasmid hydrodynamic radius increases from 116 nm to > 1300 nm and ca. 1000 nm, when formulated with CaCl2/t-butanol and aluminium phosphate gel, respectively. Circular dichroism showed that addition of CaCl2/t-butanol leads to a B→ψ (-) transition in the plasmid structure, whereas no detectable transitions were observed for aluminium p...
Source: Biotechnology and Applied Biochemistry - February 19, 2009 Category: Biochemistry Authors: M L. Wu, S S. Freitas, G A Monteiro, D M. F. Prazeres, J A. L. Santos Source Type: journals

Development of K562 cell clones expressing {beta}-globin mRNA carrying the {beta}039 thalassemia mutation for the screening of correctors of stop codon mutationsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Nonsense mutations, giving rise to UAA, UGA, UAG stop codons within the coding region of mRNAs, promote premature translational termination and are the leading cause of about 30% of inherited diseases, including cystic fibrosis, Duchenne muscular dystrophy, thalassemia. For instance, in the β039-thalassemia the CAG (Gln) codon is mutated to the UAG stop codon, leading to premature translation termination and to mRNA destabilization through the well described nonsense-mediated mRNA decay (NMD). In order to develop an approach facilitating translation and, therefore, protection from NMD, aminoglycoside antibiotics ha...
Source: Biotechnology and Applied Biochemistry - February 13, 2009 Category: Biochemistry Authors: F Salvatori, V Cantale, G Breveglieri, C Zuccato, A Finotti, N Bianchi, M Borgatti, G Feriotto, F Destro, A Canella, L Breda, S Rivella, R Gambari Source Type: journals

Molecular responses of phospholipids of Taxus cuspidata to hydrodynamic shear stress in bubble columnsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Molecular responses of phospholipids of T. cuspidata cell to hydrodynamic shear stress were investigated by using two bubble columns with different nozzle sizes. T. cuspidata cells cultured in bubble columns responded with an increase of cell membrane permeability when compared with shake flask culture, corresponding to the alteration of phospholipid constitutions. Phospholipid differences between bubble columns and shake flask cultures were analyzed by using LC/ESI-MS coupled with principle component analysis (PCA). It was found that phosphatidic acid (PA) is a main molecular species of potential biomarkers. The PA quanti...
Source: Biotechnology and Applied Biochemistry - February 9, 2009 Category: Biochemistry Authors: C Zhong, S Yang, Y Yuan Source Type: journals

Efficiently enhancing regioselective acylation of 5-azacytidine catalyzed by Candida antarctica lipase B with co-solvent mixtures as the reaction mediaEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
A comparative study of regioselective acylation of 5-azacytidine catalyzed by Candida antarctica lipase B (CAL-B) in co-solvent mixtures and in pure organic solvents was made for the first time, and it was showed that a remarkable enhancement in the initial rate and substrate conversion of the reaction could be achieved with co-solvents as the reaction media instead of pure organic solvents. For CAL-B-catalyzed regioselective synthesis of 5'-O-butyryl-5-azacytidine in co-solvent mixtures, the optimal co-solvent mixture, initial water activity, enzyme dosage, the molar ratio of vinyl butyrate to 5-azacytidine and temperatur...
Source: Biotechnology and Applied Biochemistry - January 19, 2009 Category: Biochemistry Authors: H Wu, M Zong, X Chen Source Type: journals

Screening of the endophytic fungus, Phomopsis species, for the production of Taxol, an anticancer drugEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Three different strains of the endophytic fungus, Phomopsis species, were isolated from the healthy leaves of Taxus cuspidata, Ginkgo biloba and Larix leptolepis, and screened for the production of taxol on modified liquid medium for the first time. The presence of taxol was confirmed by spectroscopic and chromatographic methods of analysis. The amount of taxol produced by this fungus was quantified by high performance liquid chromatography. The maximum amount of fungal taxol production was recorded as 418 µg/l in the strain BKH 35. The production rate was increased to 8.36 x 103 fold than that found in the culture ...
Source: Biotechnology and Applied Biochemistry - December 10, 2008 Category: Biochemistry Authors: R Senthil Kumaran, B Hur Source Type: journals

Mechanism of metastasis inhibition by cartilage polysaccharide in breast cancer cellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
As abundant amount of porcine cartilage were discarded as waste in our daily life, it is necessary to find a new usage for it. We extracted polysaccharide from cartilage and performed in vitro and in vivo experiments in cancer cells using it. Mice breast cancer pulmonary metastasis model were set up and we tried to find the mechanism of metastasis inhibition by cartilage polysaccharide. Inhibitory rate of tumor weight and inhibitory rate of metastasis indicated that cartilage polysaccharide can obviously inhibit the metastasis of breast cancer cells. The contents of LNR1, αvβ3 integrin and MMP-9 showed signif...
Source: Biotechnology and Applied Biochemistry - December 4, 2008 Category: Biochemistry Authors: A Liu, Y Hu, C Liu, X Yao, G Zhang Source Type: journals

shRNA mediated silencing of Fucosyltransferase 8 in CHO cells for the production of antibodies with enhanced antibody immune effector functionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Antibody-producing Chinese hamster ovary cells (CHO-DG44) were converted into cells producing antibodies with strongly enhanced antibody dependent cellular cytotoxicity (ADCC) by knocking down α1,6 fucosyltransferase (FuT8) via constitutive expression of shRNA against FuT8. After introduction of a FuT8 shRNA expression plasmid under the control of a U6 promoter CHO-DG44 clones with less than 5% residual FuT8 mRNA expression were isolated by selection for neomycin-resistance followed by l-NGFR-enrichment and selection for Lens culinaris agglutinin (LCA)- resistance . The CHO-DG44 clones identified produced highly afu...
Source: Biotechnology and Applied Biochemistry - November 26, 2008 Category: Biochemistry Authors: V Beuger, K Künkele, H Koll, A Gärtner, M Bähner, H Burtscher, C Klein Source Type: journals

Comparison of UriSed and IQ200 full-automatic urine sediment analyzer with manual urine analysisEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The objective of this study is to compare the performances of two urine sediment analyzers, UriSed and IQ200, with the KOVA® method for urine measurement, while evaluating the results in terms of similar parameters (cells or particles/LPF or HPF). The results indicate that the UriSed and IQ200 techniques were more reproducible (7.1-30.2%, 14.9-35.4%, respectively) than the manual technique (17.9-44.4%). Significant correlations were established among the three techniques in the evaluation of leukocytes, erythrocytes and epithelial cells. Although the UriSed, IQ200 and visual microscopic measurements were in agreemen...
Source: Biotechnology and Applied Biochemistry - November 21, 2008 Category: Biochemistry Authors: O Kadir Akin, M A. Serdar, Z Cizmeci, O Genc, S Aydin Source Type: journals

Tobacco BY-2 cells as effective bioreactor for the production of puroindolinesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Puroindolines are two small proteins so called for the presence of an hydrophobic tryptophan-rich domain. Associated to wheat starch granules in Triticum aestivum, puroindolines have been shown to be responsible for the softness of the wheat endosperm. Moreover, have been proved to possess bactericide and anti-fungal properties together with the capacity of forming very stable foams. All these features make puroindolines very attractive for medical, pharmaceutical and food industrial applications. The aim of this study was to explore a plant molecular farming approach for producing a recombinant puroindolines. Three specif...
Source: Biotechnology and Applied Biochemistry - November 18, 2008 Category: Biochemistry Authors: A Sorrentino, M Iannaccone, D Palumbo, R Capparelli, R Porta, L Mariniello Source Type: journals

Covalent immobilization of TEV-protease: a useful tool for cleavage of the His-tag of recombinant proteinsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Addition of tags (as His-tags) is extremely helpful for the affinity-purification of recombinant proteins. In several cases these tags must be removed before performing functional and structural studies. The enzyme most frequently used to cleave tags of recombinant proteins is the Tobacco Etch Virus NIa protease (TEV-protease). The continuous production of this enzyme in soluble form is a quite expensive process and not easily accessible to many laboratories. Thus, an interesting alternative is the use of TEV-protease in an immobilized form, which may be reutilized several times. The main objective of the present study was...
Source: Biotechnology and Applied Biochemistry - October 21, 2008 Category: Biochemistry Authors: A Cristina Puhl, C Inés Giacomini, G Irazoqui, F Batista-Viera, A Villarino, H Terenzi Source Type: journals

Filter preconditioning enables representative scaled-down modeling of filter capacity and viral clearance by mitigating the impact of virus spike impuritiesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Endogenous and adventitious virus removal by size-exclusion membrane filtration is a critical dedicated step in an overall viral clearance strategy employed by biologics manufacturers as required by industry regulators. However, the addition of impurities from virus spike preparations used in validation studies can significantly reduce filter capacity, resulting in an over-sized and suboptimal virus filtration step. The hydraulic filter performance and virus retention observed in conventional scaled-downed validation models may not necessarily represent performance observed during process development, nor be predictive of ...
Source: Biotechnology and Applied Biochemistry - October 10, 2008 Category: Biochemistry Authors: N Z. Khan, J J. Parrella, P W. Genest, M S. Colman Source Type: journals

Head-to-tail fusions of camelid antibodies can be expressed in planta and bind in rumen fluid.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We have compared the accumulation of recombinant variable heavy chain portions (VHH) of camelid antibodies in a variety of subcellular compartments produced in planta. The VHH coding sequences were optimised for expression in Arabidopsis thaliana and placed individually or as fused, tandem, hetero dimers into synthetic plant organelle targeting cassettes designed to target the protein to either the cytoplasm, endoplasmic reticulum (ER), protein storage vacuole or chloroplast. Accumulation of individual VHHs was only detected in plants transformed with the ER targeting cassette whereas accumulation of the tandem VHHs was de...
Source: Biotechnology and Applied Biochemistry - October 10, 2008 Category: Biochemistry Authors: S Winichayakul, A Pernthaner, R Scott, R Vlaming, N Roberts Source Type: journals

Matrix-guided cartilage regeneration in chondral defectsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Small cartilage defects treated with microfracture show comparable clinical results to those treated with autologous chondrocyte implantation (ACI). Unfortunately the regenerative tissue generated by microfracture treatment does not exhibit the structure and function of hyaline articular cartilage. Adult mesenchymal stem cells are supposed to be able to differentiate according to mechanical stimuli e.g. into chondrocytes thus generating hyaline cartilage. So far the used treatments such as the microfracture procedure are only able to induce mechanically insufficient fibrocartilage. The purpose of this study was to test the...
Source: Biotechnology and Applied Biochemistry - September 16, 2008 Category: Biochemistry Authors: B Wegener, F M. Schrimpf, M F. Pietschmann, S Milz, M Berger-Lohr, P Bergschmidt, V Jansson, P E Müller Source Type: journals

Antibacterial and antioxidant activity of the secondary metabolites from in vitro cultures of Drosera aliciaeEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The objective of this research was to evaluate antioxidant as well as antibacterial properties of secondary metabolites obtained from Drosera aliciae plants grown in vitro and examine the mechanism of their antimicrobial action. Bactericidal activity of extracts from D. aliciae as well as pure ramentaceone (naphthoquinone) present in this plant were examined against human pathogenic strains both resistant and susceptible to antibiotics. Chloroform extract proved to be more effective than methanol preparation against all of the tested strains except for Pseudomonas aeruginosa isolates with the lowest minimal bactericidal co...
Source: Biotechnology and Applied Biochemistry - September 9, 2008 Category: Biochemistry Authors: A Krolicka, A Szpitter, M Maciag, E Biskup, E Gilgenast, G Romanik, M Kaminski, G Wegrzyn, E Lojkowska Source Type: journals

Metastatic cell detection using phage-peptide modified light addressable potentiometric sensorEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Detection of metastatic cells is clinically demanded to diagnose metastasis in the early stage and access the therapeutic response to anti-cancer drugs. We applied phage display technology to cultured cells with different metastasis potential and obtained four metastasis associated peptides. The association between peptides and metastatic cells were validated by ELISA as well as biosensor studies. The selected phage-peptides not only bind SW620, the metastatic cell against which the peptides were screened, but also able to capture breast cancer cells of high metastasis. The phage-peptide modified LAPS was able to distingui...
Source: Biotechnology and Applied Biochemistry - September 4, 2008 Category: Biochemistry Authors: H Zhang, X Li, Y Bai, R Niu, Y Jia, C Zhang, L Zhang, X Feng, Y Cao Source Type: journals

Influence of carbon sources on recombinant human growth hormone production by Pichia pastoris is dependent on phenotype: Comparison of Muts and Mut+ strainsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study would be helpful to optimize the expression of other therapeutic proteins by P.pastoris. (Source: Biotechnology and Applied Biochemistry)
Source: Biotechnology and Applied Biochemistry - August 29, 2008 Category: Biochemistry Authors: M Ali Orman, P Çalik, T H. Özdamar Source Type: journals

Expression of thymosin {alpha}1 concatemer in transgenic tomato fruitsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Thymosin alpha 1(Tα1), an immune booster, plays an important role in the maturation, differentiation, and function of T cells, it also can activate the production of cytokines in dendritic cells (DCs). Tα1 is one of two thymosin proteins for potential future clinical applications. In order to express Tα1 protein in plant, we designed and synthesized the Tα1 gene according to the plant codon usage bias, and created a novel 4×Tα1 concatemer (four copies of Tα1 gene arranged end-to-end in tandem, designated 4×Tα1). Subsequently, a plant binary expression vector PG...
Source: Biotechnology and Applied Biochemistry - August 27, 2008 Category: Biochemistry Authors: Y Chen, A Wang, L Zhao, G Shen, L Cui, K Tang Source Type: journals

Microbial N-demethylation: biotransformation and recovery of a drug metaboliteEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Thirty-nine microbes were screened for the ability to selectively N-demethylate (3R,5S,E)-7-(4-(4-fluorophenyl)-6-isopropyl-2-(methyl(1-methyl-1H-1,2,4-triazol-5-yl)aminopyrimidin-5-yl)-3,5-dihydroxyhept-6-enoic acid (I), a potential drug to lower blood cholesterol levels that act by inhibiting hydroxymethyl glutaryl CoA (HMG-Co A) reductase, a key enzyme involved in cholesterol biosynthesis. Two Streptomyces species were found to carry out the desired N-demethylation. Bioconversion of compound I to compound II by S. griseus ATCC 13273 gave 80% conversion at the end of the biotransformation process. After removal of cells,...
Source: Biotechnology and Applied Biochemistry - August 13, 2008 Category: Biochemistry Authors: B L Davis, M S Liu, R L. Hanson, W L Parker, R N Patel Source Type: journals

A quantitative human monoclonal antibody immunoassay using anti-idiotypic antibody as membrane antigen surrogate with SPR detectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study describes a quantitation assay method to a therapeutic human monoclonal antibody hMAb-1 that recognizes a cell surface protein employing an anti-idiotypic antibody (anti-ID) to hMAb-1 as an surrogate antigen in an immunoassay format using SPR technology. This assay is applicable for quantitation of hMAb-1 in process streams, final drug products quality control, as well as low concentration drug substances in IV bags. The surrogate nature of the anti-ID was confirmed by demonstrating that the anti-ID displaced the interaction between the hMAb-1 and its membrane antigen in a FACS titration test. The assay format i...
Source: Biotechnology and Applied Biochemistry - August 5, 2008 Category: Biochemistry Authors: R B Wong, A Lee, V S Rangan, K Cheng Source Type: journals

Antifungal proteins production in Maize suspension culturesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The growing emergency due to the microorganisms drug resistance phenomenon, has pushed forward the search for new potential drugs alternative to those already in use. Plants represent a suitable source of new antifungal molecules, as they produce a series of defensive proteins. Among them are the Pathogenesis Related proteins (PRPs) shown effective in vitro against human pathogens. An optimized and established cell suspension culture of Zea mays was shown to constitutively secrete in the medium a series of PRPs comprising the antifungal protein zeamatin (P33679) with a final yield of around 3 mg per liter. The in vitro pro...
Source: Biotechnology and Applied Biochemistry - July 30, 2008 Category: Biochemistry Authors: F Perri, S Della Penna, F Rufini, M Patamia, M Bonito, L Angiolella, A Vitali Source Type: journals

Expression and immunogenicity of recombinant VP1 of human hepatitis A virusEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We describe secretory expression and immunogenicity of the recombinant Hepatitis A virus (HAV) VP1 from stably transformed Drosophila melanogaster S2 cells. Southern blot analysis indicated that transformed S2 cells contained multiple copies of the HAV VP1 gene in the genome. Recombinant VP1 was secreted into a culture medium with a molecular weight of 42 - 49 kDa. A maximum production level of 6.24 mg/l of recombinant VP1 was obtained in a T-flask culture of Drosophila S2 cells 5 days after induction with 0.5 mM CuSO4. The recombinant HAV VP1 protein elicited the production of specific IgA in the small intestine by oral i...
Source: Biotechnology and Applied Biochemistry - July 25, 2008 Category: Biochemistry Authors: J Lee, H Lee, J Hwang-Bo, D Shon, W Kim, I Chung Source Type: journals

Structural and biological characterizations of a novel acutobin-like enzyme isolated from the snake venom of Deinagkistrodon acutusEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Synopsis: As previously reported, a serine proteinase was purified from the snake venom of Deinagkistrodon acutus using a combination of affinity and ion exchange chromatography, and its high fibrinogen-clotting activity (2025 NIH unit/mg), indicated that this protein may have great potential as a drug to treat thrombolysis. In order to systemically determine the purified protein’s structure and activity, the protein was characterized using the following methods: mass spectrometry, isoelectric focusing, deglycosylation analysis, amino acid composition analysis, peptide mass fingerprinting, N-terminal amino acid sequ...
Source: Biotechnology and Applied Biochemistry - July 22, 2008 Category: Biochemistry Authors: Y Xin, D Dong, D Chen, R Li Source Type: journals