characterization of single domain antibodies with an engineered disul fi de bond

... strongest evidence of the enhancing role of antibodies can be inferred from the occurrence of severe dengue in infants less than year of age born to dengue-immune mothers, of severe dengue during ... Prevention of Dengue With the wide geographical spread of multiple serotypes of DENV and the potentially fatal risk of DENV infections, there is an urgent need to control the spread of DENV and design ... variable domain (VL domain) and a single constant domain (CL domain) The complementarity-determining regions (CDRs) of the antibody are made up of six loops, three of which are in each of the VH and...

Ngày tải lên: 09/09/2015, 08:16

... Sugio S, Murakami K & Yamanouchi K (2003) Kinetic analysis of enhanced thermal stability of an alkaline protease with engineered twin disul de bridges and calcium-dependent stability Biotechnol ... constants of unfolding were found to decrease in an exponential manner with increasing amounts of isopropanol At 20% (v ⁄ v) isopropanol, no further decrease of the observed rate constants of unfolding ... Mansfeld J, Vriend G, Dijkstra BW, Veltman OR, Van den Burg B, Venema G, Ulbrich-Hofmann R & Eijsink VG (1997) Extreme stabilization of a thermolysin-like protease by an engineered disul de bond...

Ngày tải lên: 19/02/2014, 17:20

... [3] Design of disulde bonds Production and purication of disulde-bonded proteins Engineering of a new disulde bond into eIGPS must take into account the presence of three parental cysteines within ... oxidative closure of the engineered (3189) disulde bond SDS PAGE in absence of mercaptoethanol Lane A, puried and spontaneously oxidized, variant (3189); lane B, as in (A), but with dithiothreitol ... with an excess of Ellmans reagent (Nbs2 [32]), leading to a single, but faster migrating band (Fig 2C) SDS micelles decorated with disulde-bonded proteins have a smaller hydrodynamic volume than...

Ngày tải lên: 24/03/2014, 03:21

... The mean of the values of three independent experiments is shown with standard errors (C) The pH-dependence of the ligation process Upper panel shows SDS ⁄ PAGE analysis of the pH-dependence of ... resulted in a time-dependent enhancement of the 57-kDa band with a concomitant diminution of the 38-kDa and 19-kDa bands, as analyzed by SDS ⁄ PAGE (Fig 3A) The 38-kDa and 19-kDa bands did not disappear ... (upper panel) and nondenaturing PAGE (lower panel) analyses of the E78H ⁄ D164N mutant and related proteins Lanes a, the purified 38-kDa mature form; lanes b, the isolated 19-kDa propeptide; lanes...

Ngày tải lên: 19/02/2014, 07:20

... isolation of singledomain antibodies (sdAbs) consisting of the variable domain of these heavy chain antibodies, also named VHH [2] These minimal antibody domains are endowed with a large number of properties ... contain an additional disul de bond and not exceed the mean CDR3 length of classical VH, in contrast to most camel VHH [18] Of note is that that none of these antibodies binds to one of the Gold ... Jalali-Javaran M, Rasaee MJ, Rahbarizadeh F, Forouzandeh-Moghadam M & Memari HR (2007) Production and characterization of anti-(mucin MUC1) single- domain antibody in tobacco (Nicotiana tabacum...

Ngày tải lên: 23/03/2014, 05:22

... , and edge set E Denote by d(v) the degree of v, where v ∈ V is a vertex of T The Laplacian matrix of T (also known to be the difference of the diagonal matrix of vertex degrees D(T) and the ... sequence with N < N nucleotides to the original sequence with N nucleotides and report the positions of the nucleotide mutations within the sequence, leading to that transition At the completion of ... secondary structure example depicted in Figure 2.1 Laplacian representation of case study The eigenvalues of the Laplacian matrix are independent of the chosen labeling for the nodes in the tree graph,...

Ngày tải lên: 22/06/2014, 23:20

Ngày tải lên: 05/08/2014, 15:20

... domain, which made it an integral membrane protein The apparent preponderance of horizontal gene transfer in the evolution of the KAP family and the phylogenetic affinities of the animal KAP NTPases ... clade that includes animal and bacterial proteins In the schematics of protein and gene structure, conserved operons are shown as boxed arrow, and transmembrane regions inserted into the KAP domain ... NTPase domain of the KAP family contains an α-helix amino-terminal of the Walker A strand and an α-helical extension with three to four predicted helical segments occurring carboxy-terminal of strand...

Ngày tải lên: 09/08/2014, 20:20

... Sensitivity analysis In a sensitivity analysis of an ISM, the selection of the quantities of interest depends on the aim of the analysis The purpose of the current analysis is to determine the order of ... the importance of using models as tools to assist the policy analysis These models can be referred to as policy analysis models (Miser and Quade, 1985) or rapid assessment models (De Kok and Wind, ... conceptual guidelines provided by Miser and Quade (1985) and Randers (1980), six steps can be distinguished in the policy analysis using integrated systems modelling to support management (De Kok and Wind,...

Ngày tải lên: 06/11/2012, 10:35

... that of MHCE, and not MLCE (Fig 8B, lane 12) Comparison of specific activities of ZHE1, MHCE and MLCE judged by synthetic peptide substrates Cleaving efficiencies of ZHE1, MHCE and MLCE were quantitatively ... enzyme of zebrafish K Sano et al channels After incubation with rec ZHE1, the fibrous structure of the inner layer became evident, and its thickness was increased two-fold more than that of the ... min, bands with molecular masses of 43 and 39 kDa were observed in addition to undigested bands of ZP2 and ZP3 After incubation for 10 min, three major bands with molecular masses of 43, 39 and...

Ngày tải lên: 07/03/2014, 04:20

... involvement of disul de bond formation We hypothesized that bacitracin may react with cysteines on PDI to form a disul de bond Using MALDI-TOF ⁄ TOF MS analysis, we were able to show that disul de bond ... isolated analogs was confirmed by MALDI-TOF ⁄ TOF MS, with a 4800 MALDITOF ⁄ TOF Analyzer (AB SCIEX, Framingham, MA, USA) MALDI-TOF and TOF ⁄ TOF MS spectra were acquired with 800–1000 and 2000–2800 ... relationship of the metalloantibiotic peptide bacitracin J Inorganic Biochem 91, 46–58 24 Wang C, Chen S, Wang X, Wang L, Wallis AK, Freedman RB & Wang CC (2010) Plasticity of human protein disul de isomerase:...

Ngày tải lên: 14/03/2014, 23:20

... Fig – XPS Spectra of WO3 nanostructures: a) O1 s and b) W4 f axis with an inter-planar distance of 0.38 nm Raman, EDS and XPS analysis confirmed that the chemical structure and oxidation states ... as the detector; and a computer-controlled XY stage with a spatial resolution of 0.1 µm with two interchangeable gratings (600 and 1800 g mm− 1) and a confocal microscope with 10, 50, and 100× ... direction of the tungsten oxide nanostructures is along [010] axis with an inter-planar distance of 0.38 nm (Fig 4b) It seems that the WO3 growth proceeds layer by layer increasing the thickness and...

Ngày tải lên: 20/03/2014, 13:08

... the same disul de linkage as mr3a and tx3c [8] Recently, a third disul de bond arrangement within M-superfamily conotoxins has been characterized Two M-1 branch conotoxins, mr3e (Fig 1) and tx3a ... conotoxins, mr3a and tx3c, have shown that they possess a distinctive disul de bond arrangement of C1–C6, C2–C4 and C3–C5 [3] In addition, BtIIIB, another M-2 branch conotoxin from the venom of a vermivorous ... spectrum Six u angle constraints and three disul de bonds from Cys2 to Cys14, Cys3 to Cys12, and Cys8 to Cys15 were added to the distance constraints for primary structure determination A set of 20 structures...

Ngày tải lên: 30/03/2014, 09:20

... electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A Anal Biochem ... produce anti-IEP IgG, the IEP-1 cDNA, with an N-terminal His6 tag, was expressed in E coli for use as an antigen This antibody detects IEP-1 and IEP-2 well by immunoblot analysis IEP is an CkPDV ... wasp reproductive tracts fixed with Carnoy’s fixative were embedded in paraffin, and 5-lm thick sections were pretreated with 5% nonfat dry milk, incubated with anti-IEP IgG or anti-(CkPDV capsid protein)...

Ngày tải lên: 31/03/2014, 15:20

... respondents were asked how often they face tight deadlines, how often they have to work at fast pace and how often they can change their pace of work or the order of their tasks (Béjean and Sultan-Taïeb, ... long derived, used and validated such an index We used the Mediterranean Diet Score index developed by Trichopoulou et al (2003) (more details on the construction and validity of the index are ... is defined as job strain (Béjean and Sultan-Taïeb, 2005, Karasek, 1979, Karasek and Theorell, 1990) Job strain occurs when job demands are high and job decision latitude is low High job demands...

Ngày tải lên: 08/04/2014, 16:55

... not allow an association of the area of fluorescence with the level of the lumbar spine The Fusion of OI data with X-rays allowed an accurate anatomical localization of the enhancing granulation ... dose of mg/kg The dose of 10 mg/ kg of ICG provided a stronger and prolonged enhancement of the granulation tissue thus allowing for longer observation times and improved detection of disease Of ... surgery in a rat model We determined the best timing and dose of an FDA-approved contrast agent that provided an optimal detection of postoperative granulation tissue on OI/X-ray images and then compared...

Ngày tải lên: 18/06/2014, 15:20

Ngày tải lên: 05/08/2014, 09:46

... different types of integration within the framework of Integrated Assessment Modelling (IAM) These are integrations of disciplines, of models, of scales, of issues, and of stakeholders Scrase and Sheate ... the importance of using models as tools to assist the policy analysis These models can be referred to as policy analysis models (Miser and Quade, 1985) or rapid assessment models (De Kok and Wind, ... conceptual guidelines provided by Miser and Quade (1985) and Randers (1980), six steps can be distinguished in the policy analysis using integrated systems modelling to support management (De Kok and Wind,...

Ngày tải lên: 10/08/2014, 01:22

... was determined using an ANOVA with Student's t test Null-hypothesis probabilities of p < 0.05 were considered significant All values are expressed as means ± SD Immunostaining of Gb3 Mice were anesthetized ... microscopy can also be seen with minimal change in disease Clinical studies of ERT for Fabry disease have demonstrated different degrees of clearance of glycosphingolipid deposits This clearance results ... achieved with a recombinant AAV2/8 vector than with an AAV2/2 vector, and that this led to significantly greater and more rapid reduction of lysosomal storage of Gb3 in the kidneys of treated Fabry...

Ngày tải lên: 10/08/2014, 05:21

... as the inverse of the highest serum dilution with a final value of optical absorbance equal to two times blank absorbance plus times the SD Response is provided as the mean of antibodies titers ... good antibody responders (GAR) if they reached anti-EGF antibody titers equal or higher than 1:4, 000 sera dilution, and super good antibody responders (SGAR) if patients reached anti-EGF antibody ... as Good Antibody Responders (GAR) if they reached an anti-EGF antibody titer ≥ 1:4000 and SGAR if they reached an anti-EGF antibody titer ≥ 1:64000 high dose/multiple sites study the decay below...

Ngày tải lên: 11/08/2014, 08:21