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antibody phage display

Yubao Cui, Lili Yu, Ying Zhou, Li Yang, Chengbo Zhang
Mimotope mapping enables the characterization of allergen epitopes for the development of diagnostic and therapeutic approaches. In the present study, a phage display peptide library was used for mimotope mapping based on the binding of antibodies against the recombinant group 5 allergen from the house dust mite Dermatophagoides farinae (Der f 5), an arthropod that causes indoor allergies worldwide. When three monoclonal anti‑Der f 5 antibodies were used for biopanning, seven mimotopes were identified. Their common subsequence was '‑‑‑[‑A][‑T]W[‑S]H[HSFW][LM][PSKR] [TLV][AST]‑[DP][‑L]‑'...
October 13, 2016: Molecular Medicine Reports
Ricardo Jara-Acevedo, Paula Díez, María González-González, Rosa María Dégano, Nieves Ibarrola, Rafael Góngora, Alberto Orfao, Manuel Fuentes
The selection process aims sequential enrichment of phage antibody display library in clones that recognize the target of interest or antigen as the library undergoes successive rounds of selection. In this review, selection methods most commonly used for phage display antibody libraries have been comprehensively described.
October 7, 2016: Current Pharmaceutical Design
Stefano Mancini, Carmen Menzi, Frank Oechslin, Philippe Moreillon, José Manuel Entenza
Streptococcus gordonii and related species of oral viridans group streptococci (VGS) are common etiological agents of infective endocarditis (IE). We explored vaccination as a strategy to prevent VGS- IE, using a novel antigen presenting system based on non-genetically modified Lactococcus lactis displaying vaccinogens on its surface.Hsa and PadA are surface-located S. gordonii proteins implicated in platelet adhesion and aggregation, which are key steps in the pathogenesis of IE. This function makes them ideal targets for vaccination against VGS- IE...
October 10, 2016: Infection and Immunity
M Brasino, J N Cha
Here we describe novel covalent conjugates of antibody-phage for the detection of multiple cancer biomarkers using real time immuno-polymerase chain reaction (immuno-PCR). While the conventional process of immuno-PCR utilizes DNA-conjugated antibodies, chemical modification of antibodies not only reduces antibody affinity but also creates a heterogeneous population of products. However, phage naturally encapsulate genomic DNA, which can be used as a PCR template. To produce covalently conjugated antibody-phage constructs without recombinant antibody expression or chemical modification of antibodies, we incorporated a photocrosslinkable non-canonical amino acid within an antibody-binding domain displayed on one of the phage coat proteins...
October 12, 2016: Analyst
William J J Finlay, Laird Bloom, Sreeja Varghese, Bénédicte Autin, Orla Cunningham
High-affinity, highly specific binding proteins are a key class of molecules used in the development of new affinity chromatography methods. Traditionally, antibody-based methods have relied on the use of immunoglobulins purified from immune animal sera, from egg yolks, or from murine monoclonal hybridoma supernatants. To accelerate and refine the reagent antibody generation process, we have developed optimized methods that allow the rapid assembly of scFv libraries from chickens immunized with pools of immunogens...
2017: Methods in Molecular Biology
William J J Finlay, Laird Bloom, Joanne Grant, Edward Franklin, Deirdre Ní Shúilleabháin, Orla Cunningham
Antibodies are critical reagents in many fundamental biochemical methods such as affinity chromatography, enzyme-linked immunosorbent assays (ELISA), flow cytometry, western blotting, immunoprecipitation, and immunohistochemistry techniques. As our understanding of the proteome becomes more complex, demand is rising for rapidly generated antibodies of higher specificity than ever before. It is therefore surprising that few investigators have moved beyond the classical methods of antibody production in their search for new reagents...
2017: Methods in Molecular Biology
Jun-Haeng Cho, A-Ru Kim, Sang-Heon Kim, Su-Jae Lee, Hoeil Chung, Moon-Young Yoon
CD133 is known as biomarker for glioblastoma (GBM) and also serves as a marker for cancer stem cells (CSCs), which carry out tumorigenesis and resist conventional therapeutics. The presence of CD133-presenting CSC is a one of the factors in maintenance of the tumorigenic potential of GBM. Thus, CD133 is a potential target for accurate diagnosis of GBM, which could improve its poor prognosis for patients when CSCs are present. Herein we designed a small peptide-based imaging agent with stimulus-responsive properties...
October 6, 2016: Acta Biomaterialia
Gene Kurosawa, Mariko Kondo, Yoshikazu Kurosawa
When the technology for constructing human antibody (Ab) libraries using a phage-display system was developed, many researchers in Ab-related fields anticipated that it would be widely applied to the development of pharmaceutical drugs against various diseases, including cancers. However, successful examples of such applications are very limited. Moreover, researchers who utilize phage-display technology now show divergent ways of thinking about phage Ab libraries. For example, there is debate about what should be the source of VH and VL genes for the construction of libraries to cover the whole repertoire of Abs present in the human body...
October 5, 2016: Biochemical and Biophysical Research Communications
Jan Müller, Robin Reichel, Sebastian Vogt, Stefan P Müller, Wolfgang Sauerwein, Wolfgang Brandau, Angelika Eggert, Alexander Schramm
Neuroectodermal tumours are characterized by aberrant processing of disialogangliosides concomitant with high expression of GD2 or GD3 on cell surfaces. Antibodies targeting GD2 are already in clinical use for therapy of neuroblastoma, a solid tumour of early childhood. Here, we set out to identify peptides with high affinity to human disialoganglioside GD2. To this end, we performed a combined in vivo and in vitro screen using a recombinant phage displayed peptide library. We isolated a phage displaying the peptide sequence WHWRLPS that specifically binds to the human disialoganglioside GD2...
2016: PloS One
Wei Li, Hongjia Yang, Dimiter S Dimitrov
CD16A (FcγRIIIA) is an activating receptor mostly expressed on natural killer (NK) cells and monocytes/macrophages. It can mediate antibody-dependent cell-mediated cytotoxicity (ADCC) through low-affinity interaction with human immunoglobulin G (IgG) Fc. It can also mediate cell lysis if NK cells are guided by bispecific killer cells engagers (BiKEs). BiKEs showed some success in clinical trials of cancer and are promising candidate therapeutics. However, currently reported BiKEs are based on antibody fragments (scFvs) of relatively large size...
October 3, 2016: Experimental and Molecular Pathology
Jonas Zantow, Sarah Just, Ilias Lagkouvardos, Sigrid Kisling, Stefan Dübel, Patricia Lepage, Thomas Clavel, Michael Hust
Pathogen infections, autoimmune diseases, and chronic inflammatory disorders are associated with systemic antibody responses from the host immune system. Disease-specific antibodies can be important serum biomarkers, but the identification of antigens associated with specific immune reactions is challenging, in particular if complex communities of microorganisms are involved in the disease progression. Despite promising new diagnostic opportunities, the discovery of these serological markers becomes more difficult with increasing complexity of microbial communities...
October 5, 2016: Scientific Reports
Sanne A M van Lith, Ilse Roodink, Joost J C Verhoeff, Petri I Mäkinen, Jari P Lappalainen, Seppo Ylä-Herttuala, Jos Raats, Erwin van Wijk, Ronald Roepman, Stef J Letteboer, Kiek Verrijp, William P J Leenders
Diffuse gliomas are primary brain cancers that are characterised by infiltrative growth. Whereas high-grade glioma characteristically presents with perinecrotic neovascularisation, large tumor areas thrive on pre-existent vasculature as well. Clinical studies have revealed that pharmacological inhibition of the angiogenic process does not improve survival of glioblastoma patients. Direct targeting of tumor vessels may however still be an interesting therapeutic approach as it allows pinching off the blood supply to tumor cells...
September 26, 2016: Oncotarget
Daniel T Harris, Ningyan Wang, Timothy P Riley, Scott D Anderson, Nishant K Singh, Erik Procko, Brian M Baker, David M Kranz
Proteins are often engineered to have higher affinity for their ligands in order to achieve therapeutic benefit. For example, many studies have used phage- or yeast-display libraries of mutants within complementarity determining regions (CDRs) to affinity mature antibodies and T cell receptors (TCRs). However, these approaches do not allow rapid assessment or evolution across the entire interface. By combining directed evolution with deep sequencing it is now possible to generate sequence fitness landscapes that survey the impact of every amino acid substitution across the entire protein-protein interface...
September 28, 2016: Journal of Biological Chemistry
Theam Soon Lim, Soo Khim Chan
BACKGROUND: Antibody phage display is highly dependent on the availability of antibody libraries. There are several forms of libraries depending mainly on the origin of the source materials. There are three major classes of libraries, mainly the naïve, immune and synthetic libraries. METHODS: Immune antibody libraries are designed to isolate specific and high affinity antibodies against disease antigens. The pre-exposure of the host to an infection results in the production of a skewed population of antibodies against the particular infection...
September 23, 2016: Current Pharmaceutical Design
Luiz Ricardo Goulart, Paula Souza Santos, Ana Paula Carneiro, Barbara Brasil Santana, Antonio C R Valinotto, Thaise Gonçalves Araújo
BACKGROUND: The identification of combinatorial antibodies against many different targets in oncology, autoimmune, inflammatory and infectious diseases has uncovered novel strategies to control and prevent diseases' onset and progression, and represents the fastest growing market for the pharmaceutical industry. Phage Display has been successfully used in the identification of unknown targets, which combines shotgun approaches with high throughput selection schemes. METHODS: This specific review covers many aspects of combinatorial phage display technology starting from antibody selection strategies to its redesign for application purposes...
September 23, 2016: Current Pharmaceutical Design
Hyunbo Shim
BACKGROUND: Antibody phage display is a major technological platform for the generation of fully human antibodies for therapeutic purposes. The in vitro binder selection by phage display allows researchers to have more extensive control over binding parameters and facilitates the isolation of clinical candidate antibodies with desired binding and/or functional profiles. METHODS: Since the invention of antibody phage display in late 1980s, significant technological advancements in the design, construction, and selection of the antibody libraries have been made, and several fully human antibodies generated by phage display are currently approved or in various clinical development stages...
September 23, 2016: Current Pharmaceutical Design
Ema Romão, Francisco Morales-Yánez, Yaozhong Hu, Maxine Crauwels, Pieter De Pauw, Gholamreza Ghassanzadeh Hassanzadeh, Nick Devoogdt, Chloé Ackaert, Cécile Vincke, Serge Muyldermans
BACKGROUND: The discovery of functional heavy chain-only antibodies devoid of light chains in sera of camelids and sharks in the early nineties provided access to the generation of minimal-sized, single-domain, in vivo affinity-matured, recombinant antigen-binding fragments, also known as Nanobodies. METHODS: Recombinant DNA technology and adaptation of phage display vectors form the basis to construct large naïve, synthetic or medium sized immune libraries from where multiple Nanobodies have been retrieved...
September 23, 2016: Current Pharmaceutical Design
Bryan D Fleming, Mitchell Ho
Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer, yet no effective therapeutics exist. This review provides an overview of the recent development of recombinant immunotoxins for the treatment of glypican-3 (GPC3) expressing HCC. GPC3 is a cell surface heparan sulfate proteoglycan that is overexpressed in HCC, but is absent from normal adult human tissues. Treatment of HCC with anti-GPC3 immunotoxins represents a new therapeutic option. Using phage display and hybridoma technologies, three high affinity antibodies (HN3, HS20 and YP7) have been generated against GPC3...
2016: Toxins
Chi-Hsin Lee, Yu-Ching Lee, Sy-Jye Leu, Liang-Tzung Lin, Jen-Ron Chiang, Wei-Jane Hsu, Yi-Yuan Yang
: Banded krait (Bungarus multicinctus; BM), one of the major envenomation species in Taiwan, contains neurotoxic venom proteins (BM proteins), that poses a serious medical problem in tropical and sub-tropical countries. Even though horse-derived serum is an efficient therapy against snake venom, it is associated with high cost and side effects. Therefore, developing a more cost-effective alternative treatment option is highly envisaged. In this study, chickens were immunized with BM proteins, and polyclonal immunoglobulin Y (IgY) antibodies were purified from eggs...
September 23, 2016: Applied and Environmental Microbiology
Aidong Wang, Ming Cui, Hong Qu, Jiabo Di, Zaozao Wang, Jiadi Xing, Fan Wu, Wei Wu, Xicheng Wang, Lin Shen, Beihai Jiang, Xiangqian Su
The epidermal growth factor receptor (EGFR) is overexpressed in several epithelial tumors. Anti-EGFR humanized monoclonal antibodies, cetuximab and panitumumab, in combination with chemotherapy have improved the prognosis for patients with wild-type RAS tumors. To identify mimotopes of EGFR and develop mimotope-based EGFR vaccines, we screened a phage display peptide library with panitumumab. Two EGFR mimotopes P19 and P26, which could be recognized by panitumumab specifically, were isolated. To enhance the immune responses, we generated recombinant proteins of P19 or P26 fused to a heat-shock cognate protein 70 (Hsc70), and evaluated the efficacy of Hsc70-P19 and Hsc70-P26 as vaccines in vivo...
September 21, 2016: Oncotarget
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