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https://www.readbyqxmd.com/read/28336958/from-rabbit-antibody-repertoires-to-rabbit-monoclonal-antibodies
#1
REVIEW
Justus Weber, Haiyong Peng, Christoph Rader
In this review, we explain why and how rabbit monoclonal antibodies have become outstanding reagents for laboratory research and increasingly for diagnostic and therapeutic applications. Starting with the unique ontogeny of rabbit B cells that affords highly distinctive antibody repertoires rich in in vivo pruned binders of high diversity, affinity and specificity, we describe the generation of rabbit monoclonal antibodies by hybridoma technology, phage display and alternative methods, along with an account of successful humanization strategies...
March 24, 2017: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/28336957/next-generation-sequencing-enables-the-discovery-of-more-diverse-positive-clones-from-a-phage-displayed-antibody-library
#2
Wonjun Yang, Aerin Yoon, Sanghoon Lee, Soohyun Kim, Jungwon Han, Junho Chung
Phage display technology provides a powerful tool to screen a library for a binding molecule via an enrichment process. It has been adopted as a critical technology in the development of therapeutic antibodies. However, a major drawback of phage display technology is that because the degree of the enrichment cannot be controlled during the bio-panning process, it frequently results in a limited number of clones. In this study, we applied next-generation sequencing (NGS) to screen clones from a library and determine whether a greater number of clones can be identified using NGS than using conventional methods...
March 24, 2017: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/28332591/improved-lysis-efficiency-and-immunogenicity-of-salmonella-ghosts-mediated-by-co-expression-of-%C3%AE-phage-holin-endolysin-and-%C3%A9-x174-gene-e
#3
Gayeon Won, Irshad Ahmed Hajam, John Hwa Lee
Bacterial ghosts (BGs) are empty cell envelopes derived from Gram-negative bacteria by bacteriophage ɸX174 gene E mediated lysis. They represent a novel inactivated vaccine platform; however, the practical application of BGs for human vaccines seems to be limited due to the safety concerns on the presence of viable cells in BGs. Therefore, to improve the lysis efficiency of the gene E, we exploited the peptidoglycan hydrolyzing ability of the λ phage holin-endolysins to expedite the process of current BG production system...
March 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28328317/antiproliferative-and-apoptotic-effects-of-novel-anti-ror1-single-chain-antibodies-in-hematological-malignancies
#4
Leili Aghebati-Maleki, Vahid Younesi, Behzad Baradaran, Jalal Abdolalizadeh, Morteza Motallebnezhad, Hamid Nickho, Dariush Shanehbandi, Jafar Majidi, Mehdi Yousefi
Receptor tyrosine kinase-like orphan receptor (ROR) proteins are a conserved family of tyrosine kinase receptors that function in developmental processes including cell survival, differentiation, cell migration, cell communication, cell polarity, proliferation, metabolism, and angiogenesis. ROR1 has recently been shown to be expressed in various types of cancer cells but not normal cells. Pharmacokinetics and pharmacodynamics of single-chain Fragment variable (scFv) antibodies provide potential therapeutic advantages over whole antibody molecules...
April 2017: SLAS Discov
https://www.readbyqxmd.com/read/28323279/spontaneous-mutations-of-a-model-heterotrophic-marine-bacterium
#5
Ying Sun, Kate E Powell, Way Sung, Michael Lynch, Mary Ann Moran, Haiwei Luo
Heterotrophic marine bacterioplankton populations display substantive genomic diversity that is commonly explained to be the result of selective forces imposed by resource limitation or interactions with phage and predators. Here we use a mutation-accumulation experiment followed by whole-genome sequencing of mutation lines to determine an unbiased rate and molecular spectrum of spontaneous mutations for a model heterotrophic marine bacterium in the globally important Roseobacter clade, Ruegeria pomeroyi DSS-3...
March 21, 2017: ISME Journal
https://www.readbyqxmd.com/read/28319812/exploring-sequence-space-harnessing-chemical-and-biological-diversity-towards-new-peptide-leads
#6
REVIEW
Richard Obexer, Louise J Walport, Hiroaki Suga
From their early roots in natural products, peptides now represent an expanding class of novel drugs. Their modular structures make them ideal candidates for pooled library screening approaches. Key technologies for library generation and screening, such as SICLOPPS, phage display and mRNA display, give unparalleled access to tight binding peptides. Through combination with genetic code reprogramming and chemical modifications, access to more natural product-like libraries, spanning non-canonical peptide space, is readily achievable...
March 17, 2017: Current Opinion in Chemical Biology
https://www.readbyqxmd.com/read/28315327/discovery-of-a-kelch-like-ech-associated-protein-1-inhibitory-tetrapeptide-and-its-structural-characterization
#7
Satoshi Sogabe, Kotaro Sakamoto, Yusuke Kamada, Akito Kadotani, Yasunori Fukuda, Jun-Ichi Sakamoto
Keap1 constitutively binds to the transcription factor Nrf2 to promote its degradation, resulting in negative modulation of genes involved in cellular protection against oxidative stress. Keap1 is increasingly recognized as an attractive target for treating diseases involving oxidative stress, including cancer, atherosclerosis, diabetes, arthritis, and neurodegeneration. We used phage-display peptide screening to identify a tetrapeptide showing moderate binding affinity, which inhibits the interaction between Nrf2 and Keap1...
March 14, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28315215/detection-of-food-allergens-by-phage-displayed-produced-antibodies
#8
Raquel Madrid, Silvia de la Cruz, Aina García, Rosario Martín, Isabel González, Teresa García
Phage display is a powerful tool to produce recombinant antibodies against a given antigen without animal immunization. This technology employs libraries of recombinant bacteriophages that display billions of different functional antibody fragments on their surface. They are selected by panning in vitro against the target antigen in search for specific binders. In this chapter, we describe the selection of single chain variable fragment (scFv) antibodies to be used for detection of allergenic proteins from nuts in food products...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28302013/multicyclic-peptides-as-scaffolds-for-the-development-of-tumor-targeting-agents
#9
Anastasia Loktev, Uwe Haberkorn, Walter Mier
The lack of specificity of traditional cytotoxic drugs triggers the development of anticancer agents with high selectivity to tumor-specific proteins. The unveiling of target structures such as EGFR or Her2 allows the focused development of novel therapies and has strongly advanced tumor treatment. Tumor-specific high-affinity ligands can be identified by using display techniques such as phage, yeast surface, ribosome and mRNA display. These techniques enable the screening of huge libraries, consequently providing a valuable alternative to rational drug development...
March 16, 2017: Current Medicinal Chemistry
https://www.readbyqxmd.com/read/28299739/mapping-the-substrate-recognition-landscapes-of-metalloproteases-using-comprehensive-mutagenesis
#10
Colin A Kretz
Protease specificity is controlled by exosites, which capture and orient the substrate, and the active site, which binds substrate residues near the P1-P1' scissile bond and catalyzes peptide hydrolysis. Techniques used to identify critical contact points between a protease and its substrate can be time consuming and labor-intensive. Screening tools such as phage display have been revitalized with the emergence of high-throughput sequencing technology, and can be used to interrogate protease substrate specificity...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28299705/characterization-of-in-vivo-selected-bacteriophage-for-the-development-of-novel-tumor-targeting-agents-with-specific-pharmacokinetics-and-imaging-applications
#11
Jessica Newton-Northup, Susan L Deutscher
Bacteriophage (phage) display technology is a powerful strategy for the identification of peptide-based tumor targeting agents for drug discovery. Phage selections performed in vitro often result in many phage clones/peptides with similar properties and often similar sequence. However, these phage and corresponding peptides are selected, validated, and characterized outside the complicated milieu of a living animal. Thus, there is no guarantee that peptides from in vitro selections will successfully meet the requirements of an in vivo targeting compound...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28289031/downregulation-of-autolysin-encoding-genes-by-phage-derived-lytic-proteins-inhibits-biofilm-formation-in-staphylococcus-aureus
#12
Lucía Fernández, Silvia González, Ana Belén Campelo, Beatriz Martínez, Ana Rodríguez, Pilar García
Phage-derived lytic proteins are a promising alternative to conventional antimicrobials. One of their most interesting properties is that they do not readily select for resistant strains, which is likely due to the fact that their targets are essential for the viability of the bacterial cell. Moreover, genetic engineering allows the design of new "tailor-made" proteins that may exhibit improved antibacterial properties. One example of this is the chimeric protein CHAPSH3b, which consists of a catalytic domain from the virion-associated peptidoglycan (PG) hydrolase of phage vB_SauS-phiIPLA88 (HydH5) and the cell wall binding domain of lysostaphin...
March 13, 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/28287337/generation-and-characterization-of-protective-antibodies-to-marburg-virus
#13
Jeffrey W Froude, Thibaut Pelat, Sebastian Miethe, Samantha E Zak, Anna Z Wec, Kartik Chandran, Jennifer Mary Brannan, Russell R Bakken, Michael Hust, Philippe Thullier, John M Dye
Marburg virus (MARV) and Ebola virus (EBOV) have been a source of epidemics and outbreaks for several decades. We present here the generation and characterization of the first protective antibodies specific for wild-type MARV. Non-human primates (NHP), cynomolgus macaques, were immunized with viral-replicon particles expressing the glycoproteins (GP) of MARV (Ci67 isolate). An antibody fragment (single-chain variable fragment, scFv) phage display library was built after four immunogen injections, and screened against the GP1-649 of MARV...
March 13, 2017: MAbs
https://www.readbyqxmd.com/read/28281773/b-cell-epitopes-of-african-horse-sickness-virus-serotype-4-recognised-by-immune-horse-sera
#14
Evans M Mathebula, Frederika E Faber, Wouter Van Wyngaardt, Antoinette Van Schalkwyk, Alri Pretorius, Jeanni Fehrsen
Identifying antigenic proteins and mapping their epitopes is important for the development of diagnostic reagents and recombinant vaccines. B-cell epitopes of African horse sickness virus (AHSV) have previously been mapped on VP2, VP5, VP7 and NS1, using mouse, rabbit and chicken monoclonal antibodies. A comprehensive study of the humoral immune response of five vaccinated horses to AHSV-4 antigenic peptides was undertaken. A fragmented-genome phage display library expressing a repertoire of AHSV-4 peptides spanning the entire genome was constructed...
February 24, 2017: Onderstepoort Journal of Veterinary Research
https://www.readbyqxmd.com/read/28279647/isolation-of-a-peptide-from-ph-d-c7c-phage-display-library-for-detection-of-cry1ab
#15
Yun Wang, Qian Wang, Ai-Hua Wu, Zhen-Ping Hao, Xian-Jin Liu
Traditional ELISA methods of using animal immunity yield antibodies for detection Cry toxin. Not only is this incredibly harmful to the animals, but is also time-intensive. Here we developed a simple method to yield the recognition element. Using a critical selection strategy and immunoassay we confirmed a clone from the Ph.D-C7C phage library, which has displayed the most interesting Cry1Ab-binding characteristics examined in this study (Fig. 1). The current study indicates that isolating peptide is an alternative method for the preparation of a recognition element, and that the developed assay is a potentially useful tool for detecting Cry1Ab...
March 6, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28276594/structural-and-functional-characterization-of-a-ubiquitin-variant-engineered-for-tight-and-specific-binding-to-an-alpha-helical-ubiquitin-interacting-motif
#16
Noah Manczyk, Bradley P Yates, Gianluca Veggiani, Andreas Ernst, Frank Sicheri, Sachdev S Sidhu
Ubiquitin interacting motifs (UIMs) are short α-helices found in a number of eukaryotic proteins. UIMs interact weakly but specifically with ubiquitin conjugated to other proteins, and in so doing, mediate specific cellular signals. Here we used phage display to generate ubiquitin variants (UbVs) targeting the N-terminal UIM of the yeast Vps27 protein. Selections yielded UbV.v27.1, which recognized the cognate UIM with high specificity relative to other yeast UIMs and bound with an affinity more than two orders of magnitude higher than that of ubiquitin...
March 9, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28272485/construction-and-immunogenicity-of-a-recombinant-swinepox-virus-expressing-a-multi-epitope-peptide-for-porcine-reproductive-and-respiratory-syndrome-virus
#17
Huixing Lin, Zhe Ma, Xin Hou, Lei Chen, Hongjie Fan
To characterize neutralizing mimotopes, phages were selected from a 12-mer phage display library using three anti-porcine reproductive and respiratory syndrome virus (PRRSV) neutralizing monoclonal antibodies: (1) A1; (2) A2; and (3) A7. Of these, A2 and A7 recognize the mimotope, P2, which contains the SRHDHIH motif, which has conserved consensus sequences from amino acid positions 156 to 161 in the N-terminal ectodomain of GP3. The artificial multi-epitope gene, mp2, was designed by combining three repeats of the mimotope P2...
March 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28272300/a-human-antibody-that-binds-to-the-sixth-ig-like-domain-of-vcam-1-blocks-lung-cancer-cell-migration-in-vitro
#18
Mi Ra Kim, Ji Hye Jang, Chang Sik Park, Taek-Keun Kim, Youn-Jae Kim, Junho Chung, Hyunbo Shim, In Hyun Nam, Jung Min Han, Sukmook Lee
Vascular cell adhesion molecule-1 (VCAM-1) is closely associated with tumor progression and metastasis. However, the relevance and role of VCAM-1 in lung cancer have not been clearly elucidated. In this study, we found that VCAM-1 was highly overexpressed in lung cancer tissue compared with that of normal lung tissue, and high VCAM-1 expression correlated with poor survival in lung cancer patients. VCAM-1 knockdown reduced migration of A549 human lung cancer cells into Matrigel, and competitive blocking experiments targeting the Ig-like domain 6 of VCAM-1 (VCAM-1-D6) demonstrated that the VCAM-1-D6 domain was critical for VCAM-1 mediated A549 cell migration into Matrigel...
March 6, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28271636/proximity-labeling-of-interacting-proteins-application-of-bioid-as-a-discovery-tool
#19
REVIEW
Peipei Li, Jingjing Li, Li Wang, Li-Jun Di
Protein performs biochemical functions by forming complexes, or protein-protein interactions (PPIs). Many different approaches, such as phage display and yeast hybridization etc. were developed to illustrate the PPIs, and disclose the composition and organization of protein complexes. However, none of these approaches are based on the real-time and in vivo PPI analysis. Proximity dependent labeling of interacting proteins (PDL) has recently been proposed by taking advantage of several enzymes, which are capable of attaching the known reactive groups to the nearby proteins covalently...
March 8, 2017: Proteomics
https://www.readbyqxmd.com/read/28259978/ectopic-expression-of-the-atp-synthase-%C3%AE-subunit-on-the-membrane-of-pc-3m-cells-supports-its-potential-role-in-prostate-cancer-metastasis
#20
Wei Li, Yulin Li, Gaiyun Li, Zilong Zhou, Xiaona Chang, Yang Xia, Xinjie Dong, Zhijing Liu, Bo Ren, Wei Liu, Yilei Li
Metastatic prostate cancer is associated with high mortality rates. Identification of metastasis-related proteins may facilitate the development of novel therapies for the treatment of metastatic disease. In the present study, we aimed to identify prostate cancer metastasis-associated membrane proteins. We developed a phage-displayed 7-mer peptide library to screen the target peptides that were specifically bound to PC-3M cells with subtractive panning from normal prostate cells and PC-3 prostate cancer cells...
February 16, 2017: International Journal of Oncology
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