selection of human fab libraries by phage display

... 2004 Selection of folded proteins by phage- display (Eur J Biochem 271) 1611 Fig Effect of structural change on the selection (A) Structure of cyt b562 Residues M7 and H102 are the ligands of heme ... result of selection If the cutting sites are in the flexible loop of the native structure, they could prevent the selection of stable proteins By examining the structures of the proteins studied by ... versus phage- display Fig Effect of a partially unfolded intermediate on the selection result If the cutting site for protease is in the unfolded region of an intermediate state, selection of stable...

Ngày tải lên: 19/02/2014, 12:20

... Selection of IgG by tandem phage panning-FACS Y Mazor et al was the first fully human mAb discovered using phage display to receive FDA approval, and at least 16 human mAbs derived from phage ... that of scFv [31,36] To address some of the limitations associated with scFv and Fab phage display, Lee et al [36] recently reported the development of a system for the display of bivalent Fab ... mentous phage particles For the establishment of an IgG phage display system, we took advantage of the fUSE5–ZZ phage These phage particles display the Fc-binding ZZ protein on all five copies of the...

Ngày tải lên: 22/03/2014, 21:21

... Construction of the substrate phage display library Substrate phage libraries were generated using a modified pH0508b phagemid [20] The construction consists of a His6 tag at either end of a Gly-Gly-Gly-Ser-repeat-rich ... reported the extent of diversity of the library used in the screening, this being a direct product of the number of different combinations of amino acids displayed by the phage Cloning substrates ... than that of fused to a phage The CFP system developed in the present work enabled direct determination of specificity constants and the site of cleavage of the substrate selected by phage display, ...

Ngày tải lên: 31/03/2014, 23:20

... displacement of CRT from GABARAP by the binding of N1 to GABARAP, suggesting the direct competition of the peptide with CRT for a common binding site on natively folded GABARAP Discussion Phage display of ... sequence alignment of phage display selected peptides with highest similarity to N1 Phage display selected peptide N1 binds to GABARAP We quantitatively investigated the binding of fluorescein-labelled ... interactions [26] In our phage display screen against GABARAP, a large variety of similar but not identical peptide sequences were observed The observation that none of the phage display selected peptides...

Ngày tải lên: 16/03/2014, 05:20

... cleaved by hK2 [16] The current study proposes the use of peptide substrates selected by phage- display technology to change the specificity of serpin ACT which is known to inhibit a large panel of human ... Residues P3-P3¢ located in the RSL structure of rACTWT were replaced by substrate pentapeptides previously selected by phagedisplay technology [16] Six variants of rACT have been designed and constructed ... production of fully intact ACTs purified in one step by affinity chromatography The efficiency of the bacterial recombinant system to produce active ACT was proved by the stoichiometry of inhibition of...

Ngày tải lên: 30/03/2014, 13:20

... versatility of phage library design to display peptides or proteins of varying length and complexity (42, 43) The key steps in the selection of specific antibodies from phage display libraries ... homogenate of 65 infected Ae.aegypti, detected by Immunofluorescence Figure 19 Generation of hybridoma and construction of the chimeric 68 human/ mouse EDIII phage library Figure 20 Binding of 3H5 ... addition, Phage Display Technology was utilized to generate a DENV-immune Fab phage library for investigation of antibody repertoire upon infection, and for identification of EDIII-specific Fabs that...

Ngày tải lên: 13/10/2015, 16:41

... identification of transcription start site of human KCTD10 To determine the genomic structure of human KCTD10, a full-length cDNA sequence of human KCTD10 was used to search the human genome database ... Alignment of the 5¢-flanking sequences upstream of the start codon of human and mouse KCTD10 B Fig In vivo ChIP assays of Sp1.1 and AP-2.2 binding sites in the proximal promoter region of human KCTD10 ... KCTD10 R Liu et al A B Fig Genomic structure of human and mouse KCTD10 genes, and the 5¢-flanking region sequence of human KCTD10 (A) Genomic organization of human and mouse KCTD10 genes Solid boxes...

Ngày tải lên: 30/03/2014, 02:20

... cells as defined by the presence or absence of CD27 expression We studied the effect of CP-870,893 on B cell activation and B cell stimulation of T cells, and we analyzed the effects of co-stimulating ... naïve B cells can be activated by the drug CP-870,893, and this CP870,893 effect can be increased by the addition of CpG ODN 2006 Naïve B cells, as defined by lack of CD27 expression, appear relatively ... amounts of B cell CD86 expression in response to titrated amounts of the CD40 mAb CP-870,893 CD19+ B cells were purified from PBMC by negative selection and stimulated in the presence of hIgG2...

Ngày tải lên: 18/06/2014, 15:20

... effect was merely a result of doubling the total amount of IFNs per culture, we tested the inhibitory effects of 200 IU/ml of each IFN separately Two-hundred IU/ml of IFN-α, IFN-β or IFN-γ reduced ... formation by only 11-, 37- or 30fold, respectively (Table 1) The level of inhibition was not significantly greater than the level of inhibition achieved by each IFN at concentrations of 100 IU/ml ... Illustration of a representative isobologram for a combination of two drugs The solid line is the line of additivity When the isobole lies below the line of additivity, the combinatorial effect of drug...

Ngày tải lên: 18/06/2014, 22:20

... Figure Expression of TLR by HFF and Ectocervical tissue Expression of TLR by HFF and Ectocervical tissue Expression of TLR2, TLR3, TLR4 and TLR9 in cells and tissue was assessed by reverse-transcription ... anti-HCMV effect of TLR2, TLR3, TLR4, and TLR9 ligands in ectocervical tissues No interferon-α was detected in supernatants of TLR-stimulated cultures by ELISA (not shown) Expression of TLR by HFF and ... oligonucleotide) stimulate human foreskin fibroblasts (HFF) by measuring IL-8 secretion since IL-8 is secreted by a wide variety of cell types in response to stimulation by TLR ligands [21] HFF...

Ngày tải lên: 20/06/2014, 01:20

... effect was merely a result of doubling the total amount of IFNs per culture, we tested the inhibitory effects of 200 IU/ml of each IFN separately Two-hundred IU/ml of IFN-α, IFN-β or IFN-γ reduced ... formation by only 11-, 37- or 30fold, respectively (Table 1) The level of inhibition was not significantly greater than the level of inhibition achieved by each IFN at concentrations of 100 IU/ml ... Illustration of a representative isobologram for a combination of two drugs The solid line is the line of additivity When the isobole lies below the line of additivity, the combinatorial effect of drug...

Ngày tải lên: 20/06/2014, 04:20

... tracking, FIGURE Use and selection of altered habitats by Bull Sharks B2–B9 (subset of tracks where any use of altered habitat occurred) The black bars represent the proportion of observed positions ... sharks by a tether of monel wire wrapped to the stem of the rototag The transmitter trailed between the first and second dorsal fins of the shark as it swam Transmitters weighed less than 1% of the ... at one of the two outfalls on 123 of the subsequent 156 d leading up to November 2004, after which it was no longer detected (Figure 3, inset) The shark spent most of its time just off of Plant...

Ngày tải lên: 04/09/2015, 17:13

... 4.2 Induction of SULT1A3 by ligands of GR and AhR 61 4.3 Regulation of SULT1A3 by glucocorticoids via GR .63 4.4 Regulation of SULT1A3 via AhR 65 4.5 Effects of other nuclear ... contrast, induction of human SULT1A3 by nuclear receptors has not been well studied Thus, in this study, we systematically examined the induction of human SULT1A3 genes by a whole range of nuclear receptor ... The function of PPARs is modified by the precise shape of their ligand-binding domain induced by ligand binding and by a number of coactivator and corepressor proteins, the presence of which can...

Ngày tải lên: 22/10/2015, 22:37

... Law: Principle of Human Dignity versus Principle of State Sovereignty: 37 I Basis of International Law: The Principle of Sovereignty 39 II Basis of Human Rights Law: The Principle of Human Dignity ... spirit of the doctrine of human rights and human dignity at the centre of that circle The first two ends speak of achieving peace based on human rights and human dignity The third end speaks of establishing ... defining stuff of the principle of human dignity Hence; we may Article of the Statute of the International Court of Justice Article 20 of the Statute of the International Court of Justice (B)...

Ngày tải lên: 07/12/2015, 00:47

... City, CA, USA) Isolation of anti-Cn2 scFv by panning of phage- antibody repertories The library of human scFv was displayed on filamentous phage and used for the selection of antibodies against Cn2 ... for the selection of a variety of improved clones Different strategies with the same purposes have been reported [26–29] The standard procedure of phage selection gave a lower number of positive ... isolate human neutralizing antibodies L Riano-Umbarila et al ˜ Table Characterization of scFvs selected by directed evolution and phage display Results of sequence analyses allowing identification of...

Ngày tải lên: 23/03/2014, 13:20

... Panning for human Fab from a phage display library Human Fab screening was performed using a non-immunized human Fab phage display library HX01 (Humanyx Pte Ltd, Singapore) The four serotypes of Dengue ... large array of antibodies and non-immunized human phage library such as the Humanyx Fab phage display library was constructed by amplifying the V-genes from the IgM mRNA of B cells of non-immunized ... objectives of this study are to isolate unique human anti-DENV antibodies by screening all serotypes of DENV sequentially using a human non-dengue immunized phage display library (Humanxy library)...

Ngày tải lên: 13/10/2015, 16:41

... presence of a 1000fold excess of phages displaying the other protein (B) Titration by antigen (Phl p 5)-specific ELISA of polyclonal phage stocks to illustrate the enhanced recognition of allergen ... development of specific binders at a high rate are on-going [4,15,16] Large collections of binders in the form of molecular libraries intended for different selection procedures, including phage display, ... binders by phage display The selection unit is further divided into tasks to define the placement of the herein-designed separation unit in the process (B) Specific antibody fragment-displaying phage...

Ngày tải lên: 23/03/2014, 06:20

... evaluated by measuring the effect of the phage on cAMP production None of the ECL2-specific phage clones displayed agonistic properties The antagonistic properties of the selected HCDR3 phage were ... peptide and HCDR3 phage libraries Screening of positive clones Five different fully randomized phage- displayed peptide libraries were screened For minor coat protein (pIII) displayed libraries, linear ... feasibility of further reducing the size of the antibody to the peptide level by engineering phage libraries displaying heavy chain complementarity determining region (HCDR3) repertoires representing human...

Ngày tải lên: 28/03/2014, 22:20

... http://www.virologyj.com/content/5/1/130 Phage display Fab library screening Library screening was performed using the naïve human Fab phage display library HX01 (Humanyx Pte Ltd, Singapore) Antigen ... Acknowledgements Conversion of Fab- phage to human IgG1 Distinct immunoglobulin sequences were converted to IgG1 by cloning the Fab our Fab- IgG1 vector through Apa L1 and Bsm BI The Fab- IgG1 vector was ... ensured that the selection of Fabs was not directed towards epitopes on the VLPs Analysis of the Fab from 190 single colonies identified 25 positives by ELISA against INDO05-FO, all of which were...

Ngày tải lên: 20/06/2014, 01:20

... cultures of TG1(pSOF22) with helper phage stocks at a multiplicity of infection of 50 phage per bacterium Helper phages VCSM13 and VCSM13d3 were used for production of phagemid particles of the pSOF22 ... (b) Binding of the SOF22-displaying phagemid particles to human fibronectin detected by phage ELISA A total of 108 phagemid particles were used per assay, each carried out in a well of a 96-well ... vector pDJ01 This phagemid was named pSOF22 Production and functional assays of the SOF-displaying phagemid particles The phagemid particles were generated by infection of 100 ml of exponentially...

Ngày tải lên: 14/08/2014, 08:20