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antibody phage display

C Lo Passo, L Zippilli, A Angiolillo, I Costa, I Pernice, R Galbo, F Felici, P T Beernink
Factor H binding protein (FHbp) is a component of two licensed vaccines for prevention of sepsis and meningitis caused by serogroup B meningococci. FHbp binds human Factor H (FH), which contributes to evasion of host immunity and FHbp sequence variants can be classified into two sub-families. Antibodies against FHbp elicit complement-mediated killing and can inhibit recruitment of FH to the bacterial surface. We report epitope mapping studies of two murine IgG mAbs, designated JAR 31 and JAR 36, isolated from a mouse immunized with FHbp in sub-family A, which is present in ∼30-40% of invasive isolates...
April 2018: Heliyon
Beatrix Kotlan, Vanda Plotar, Klara Eles, Szabolcs Horvath, Timea Balatoni, Orsolya Csuka, Mihaly Újhelyi, Ákos Sávolt, Andras Szollar, Istvan Vamosi-Nagy, Laszlo Toth, Emil Farkas, Jozsef Toth, Miklos Kasler, Gabriella Liszkay
Aim of the study: The arsenal of questions and answers about the minor cancer initiating cancer stem cell (CSC) population put responsible for cancer invasiveness and metastases, has left with an unsolved puzzle. Specific aims of a complex project were partly focused on revealing new biomarkers of cancer. We designed and set up novel techniques to facilitate the detection of cancerous cells. Materials and methods: As a novel approach, we investigated B cells infiltrating breast carcinomas and melanomas (TIL-B) in terms of their tumour antigen binding potential...
March 2018: Contemporary Oncology Współczesna Onkologia
Ruramayi M Nzuma, Fuquan Liu, Irene R Grant
Campylobacter jejuni is a leading cause of foodborne illness worldwide, mainly due to consumption and handling of contaminated raw chicken. Rapid detection methods for C. jejuni are vital for monitoring contamination levels in chicken products and reducing human Campylobacteriosis cases. The 'gold standard' culture-based method of Campylobacter detection takes 3-5 days and is too slow to permit effective intervention. Immuno-based methods are faster, but usually necessitate use of animals or hybridoma technology to produce antibodies; making them difficult and expensive to produce...
April 7, 2018: Applied Microbiology and Biotechnology
Hideaki Sanada, Kazuki Kobayashi, Kenji Oyama, Takamitsu Maru, Takeshi Nakanishi, Mitsuo Umetsu, Ryutaro Asano, Izumi Kumagai
Affinity maturation is one of the cardinal strategies for improving antibody function using in vitro evolutionary methods; one such well-established method is phage display. To minimise gene deletion, we previously developed an open sandwich (OS) method wherein selection was performed using only phage-displaying VH fragments after mixing with soluble VL fragments. The decrease in anti-EGFR antibody 528 affinity through humanization was successfully recovered by selecting VH mutants using this OS method. However, the affinity was not similar to that of parental 528...
April 3, 2018: Scientific Reports
Marcos B Luiz, Soraya S Pereira, Nidiane D R Prado, Naan R Gonçalves, Anderson M Kayano, Leandro S Moreira-Dill, Juliana C Sobrinho, Fernando B Zanchi, André L Fuly, Cleberson F Fernandes, Juliana P Zuliani, Andreimar M Soares, Rodrigo G Stabeli, Carla F C Fernandes
Toxic effects triggered by crotalic envenoming are mainly related to crotoxin (CTX), composed of a phospholipase A₂ (CB) and a subunit with no toxic activity (CA). Camelids produce immunoglobulins G devoid of light chains, in which the antigen recognition domain is called VHH. Given their unique characteristics, VHHs were selected using Phage Display against CTX from Crotalus durissus terrificus . After three rounds of biopanning, four sequence profiles for CB (KF498602, KF498603, KF498604, and KF498605) and one for CA (KF498606) were revealed...
March 29, 2018: Toxins
Atefeh Arab, Rezvan Yazdian Robati, Jessica Nicastro, Roderick Slavcev, Javad Behravan
Despite years of investigation, breast cancer remains a major cause of death worldwide. Phage display is a powerful molecular method in which peptide and protein libraries can be displayed via genetic fusions on the surface of phages. This approach has tremendous potential for biomedical applications and has already facilitated the discovery of specific antibodies, specific antigens, and peptides with potential roles in the diagnosis and treatment of malignancies including breast cancer. In this review, we discuss the new and the latest advancements in the applications of the phage display technique in provision of immune therapeutics for breast cancer...
March 27, 2018: Current Pharmaceutical Design
Marcus Rebouças Santos, Viviane Sisdelli Assao, Fabiana de Almeida Araújo Santos, Rafael Locatelli Salgado, Ana Paula Carneiro, Juliana Lopes Rangel Fietto, Gustavo Costa Bressan, Márcia Rogéria de Almeida, Zelia Inês Portela Lobato, Carlos Ueira-Veira, Luíz Ricardo Goulart, Abelardo Silva-Júnior
Porcine circovirus 2 (PCV2) is associated with a series of swine diseases. There is a great interest in improving our understanding of the immunology of PCV2, especially the properties of the viral capsid protein Cap-PCV2 and how they relate to the immunogenicity of the virus and the subsequent development of vaccines. Phage display screening has been widely used to study binding affinities for target proteins. The aim of this study was to use phage display screening to identify antigenic peptides in the PCV2 capsid protein...
March 27, 2018: Archives of Virology
Watee Seesuay, Surasak Jittavisutthikul, Nawannaporn Sae-Lim, Nitat Sookrung, Yuwaporn Sakolvaree, Wanpen Chaicumpa
Small molecular inhibitors and passive immunization against Ebola virus disease (EVD) have been tested in animal models, including rodents and non-human primates, as well as in clinical trials. Nevertheless, there is currently no Food and Drug Administration (FDA)-approved therapy, and alternative strategies must be pursued. The aim of this study was to produce cell-penetrable human single-chain antibodies (transbodies) that are able to interfere with the activities of interferon inhibitory domain (IID) of the VP35 protein, a multifunctional virulence factor of Ebola virus (EBOV)...
March 21, 2018: Emerging Microbes & Infections
Annalee W Nguyen, Kevin C Le, Jennifer A Maynard
Discovery of monoclonal antibodies is most commonly performed using phage or yeast display but mammalian cells are used for production because of the complex antibody structure, including the multiple disulfide bonds and glycosylation, required for function. As this transition between host organisms is often accompanied by impaired binding, folding or expression, development pipelines include laborious plate-based screening or engineering strategies to adapt an antibody to mammalian expression. To circumvent these problems, we developed a plasmid-based Fab screening platform on Chinese hamster ovary (CHO) cells which allows for antibody selection in the production host and in the presence of the same post-translational modifications as the manufactured product...
March 16, 2018: Protein Engineering, Design & Selection: PEDS
Chuan Loo Wong, Chean Yeah Yong, Azira Muhamad, Amir Syahir, Abdul Rahman Omar, Chin Chin Sieo, Wen Siang Tan
Foot-and-mouth disease (FMD) is a major threat to the livestock industry worldwide. Despite constant surveillance and effective vaccination, the perpetual mutations of the foot-and-mouth disease virus (FMDV) pose a huge challenge to FMD diagnosis. The immunodominant region of the FMDV VP1 protein (residues 131-170) displayed on phage T7 has been used to detect anti-FMDV in bovine sera. In the present study, the functional epitope was further delineated using amino acid sequence alignment, homology modelling and phage display...
March 21, 2018: Applied Microbiology and Biotechnology
Hang Yang, Yanfang Zhong, Juncheng Wang, Qinghong Zhang, Xiulan Li, Sumei Ling, Shihua Wang, Rongzhi Wang
Interferon gamma (IFN-γ), a signal proinflammatory cytokine secreted by immune cell, and plays a critical role in the pathogenesis and progression of many diseases. It has been regarded as an important marker for determination of disease-specific immune responses. Therefore, it is urgent to develop a feasible and accurate method to detect IFN-γ in clinic real blood samples. Until now, the immunoassay based on singe chain variable fragment (scFv) antibody for human IFN-γ is still not reported. In the present study, an scFv antibody named scFv-A8 with high specificity was obtained by phage display and biopanning, with the affinity 2...
2018: Frontiers in Microbiology
Seung Yub Han, Alesia Antoine, David Howard, Bryant Chang, Woo Sung Chang, Matthew Slein, Gintaras Deikus, Sofia Kossida, Patrice Duroux, Marie-Paule Lefranc, Robert P Sebra, Melissa L Smith, Ismael Ben F Fofana
The simian immunodeficiency virus (SIV)/macaque model of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome pathogenesis is critical for furthering our understanding of the role of antibody responses in the prevention of HIV infection, and will only increase in importance as macaque immunoglobulin (IG) gene databases are expanded. We have previously reported the construction of a phage display library from a SIV-infected rhesus macaque ( Macaca mulatta ) using oligonucleotide primers based on human IG gene sequences...
2018: Frontiers in Immunology
V Vasilca, A Sadeghpour, S Rawson, L E Hawke, S A Baldwin, T Wilkinson, D Bannister, V L G Postis, M Rappolt, S P Muench, L J C Jeuken
Screening assays performed against membrane protein targets (e.g. phage display) are hampered by issues arising from protein expression and purification, protein stability in detergent solutions and epitope concealment by detergent micelles. Here, we have studied a fast and simple method to improve screening against membrane proteins: spherical-supported bilayer lipid membranes ("SSBLM"). SSBLMs can be quickly isolated via low-speed centrifugation and redispersed in liquid solutions while presenting the target protein in a native-like lipid environment...
March 12, 2018: Analytical Biochemistry
Fatemeh Faraji, Nader Tajik, Mahdi Behdani, Mohammad Ali Shokrgozar, Amir Hassan Zarnani, Fatemeh Shahhosseini, Mahdi Habibi-Anbouhi
CD22 is a B-cell-specific trans-membrane glycoprotein which is found on the surface of the most B-cells and modulates their function, survival, and apoptosis. Recently, targeting this cell surface biomarker in B-cell malignancies and disorders has attracted a lot of attention. The variable domain of camelid single chain antibodies (VHH, Nanobody) is a form of antibodies with novel properties including small size (15-17 kDa), thermal and chemical stability, high affinity and homology to human antibody sequences...
March 15, 2018: Biotechnology and Applied Biochemistry
Pooja Sadana, Rebecca Geyer, Joern Pezoldt, Saskia Helmsing, Jochen Huehn, Michael Hust, Petra Dersch, Andrea Scrima
Yersinia pseudotuberculosis is a Gram-negative bacterium and zoonotic pathogen responsible for a wide range of diseases, ranging from mild diarrhea, enterocolitis, lymphatic adenitis to persistent local inflammation. The Y. pseudotuberculosis invasin D (InvD) molecule belongs to the invasin (InvA)-type autotransporter proteins, but its structure and function remain unknown. In this study, we present the first crystal structure of InvD, analyzed its expression and function in a murine infection model, and identified its target molecule in the host...
March 13, 2018: Journal of Biological Chemistry
Abbas Shali, Sadegh Hasannia, Fatemeh Gashtasbi, S Masoud Abdous, S Shirin Shahangian, Shirin Jalili
Since anthrax is an acute infectious disease, detection and neutralization of the toxins of pathogenic Bacillus anthracis are of great importance. The critical role of protective antigen (PA) component of tripartite anthrax toxin in toxin entry into the host cell cytosol provided a great deal of effort to generate monoclonal antibodies against this constitute. Regarding the importance of anthrax detection/neutralization and unique physicochemical and pharmacological features of VHHs as single domain antibodies, the present study aimed to generate VHHs against the receptor binding domain of PA, termed PAD4...
March 7, 2018: International Journal of Biological Macromolecules
Mengyao Wang, Yuhua Wei, Wei Yu, Lizi Wang, Lu Zhai, Xiaoting Li, Xintong Wang, Hua Zhang, Zhenyue Feng, Liquan Yu, Yongzhong Yu, Jinzhu Ma, Yudong Cui
The GapC protein of Staphylococcus aureus (S. aureus) is a surface protein that is highly conserved among Staphylococcus strains, and it can induce protective humoral immune responses. However, B-cell epitopes in S. aureus GapC have not been reported. In this study, we generated a monoclonal antibody (mAb2A9) targeting S. aureus GapC. Through a passive immunity test, mAb2A9 was shown to partially protect mice against S. aureus infection. We screened the motif236 PVATGSLTE243 that is recognized by mAb2A9 using a phage-display system...
March 6, 2018: Microbial Pathogenesis
Marion David, Pascaline Lécorché, Maxime Masse, Aude Faucon, Karima Abouzid, Nicolas Gaudin, Karine Varini, Fanny Gassiot, Géraldine Ferracci, Guillaume Jacquot, Patrick Vlieghe, Michel Khrestchatisky
Insufficient membrane penetration of drugs, in particular biotherapeutics and/or low target specificity remain a major drawback in their efficacy. We propose here the rational characterization and optimization of peptides to be developed as vectors that target cells expressing specific receptors involved in endocytosis or transcytosis. Among receptors involved in receptor-mediated transport is the LDL receptor. Screening complex phage-displayed peptide libraries on the human LDLR (hLDLR) stably expressed in cell lines led to the characterization of a family of cyclic and linear peptides that specifically bind the hLDLR...
2018: PloS One
Sa Dong, Zongyi Bo, Cunzheng Zhang, Jianguo Feng, Xianjin Liu
Single-chain variable fragment (scFv) is a kind of antibody that possess only one chain of the complete antibody while maintaining the antigen-specific binding abilities and can be expressed in prokaryotic system. In this study, scFvs against Cry1 toxins were screened out from an immunized mouse phage displayed antibody library, which was successfully constructed with capacity of 6.25 × 107  CFU/mL. Using the mixed and alternative antigen coating strategy and after four rounds of affinity screening, seven positive phage-scFvs against Cry1 toxins were selected and characterized...
February 26, 2018: Applied Microbiology and Biotechnology
Wenjing Zhong, Guanghui Li, Xiaolu Yu, Min Zhu, Likun Gong, Yakun Wan
Bt Cry1B toxin, a residue in insect-resistant transgenic plants, has been identified to be harmful to human health. Therefore, it is urgent to detect the Cry1B toxin level in each kind of transgenic plant. Nbs, with prominently unique physiochemical properties, are becoming more and more promising tools in the detection of target antigens. In this study, an immune phage display library that was of high quality was successfully constructed for the screening of Cry1B-specific Nbs with excellent specificity, affinity, and thermostable...
February 24, 2018: MicrobiologyOpen
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