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antibody library

X-J Yu, Y-F Shen, J Dong, T Li, C Wang, Y-J Zhang, L-F Wang, Y-C Meng, Y Yang, H-J Wang, C-H Lei, S Hu, B-H Li
Previously, we have reported the crystal structures of Fab fragment of Infliximab in complex with TNFα. The structurally identified epitope on TNFα revealed the mechanism of TNFα inhibition by partially overlapping with the TNFα-receptor interface and the possibility to optimize the binding affinity. In this study, we launched a screen of a phage display library to isolate novel anti-TNFα antibodies based on the infliximab epitope. To develop novel anti-TNFα antibodies, structural analysis, the phage display antibody isolation, step by step antibody optimization, CDR residues random mutagenesis, and binding affinity characterization were performed...
July 2018: Molekuliarnaia Biologiia
Tyler Lopez, Chen Chuan, Aaron Ramirez, Kuan-Hui E Chen, Mary Y Lorenson, Chris Benitez, Zahid Mustafa, Henry Pham, Ramon Sanchez, Ameae M Walker, Xin Ge
Targeting effectual epitopes is essential for therapeutic antibodies to accomplish their desired biological functions. This study developed a competitive dual color FACS to maturate a matrix-metalloprotease 14 (MMP-14) inhibitory antibody. Epitope specific screening was achieved by selection on MMP-14 during competition with nTIMP-2, a native inhibitor of MMP-14 binding strongly to its catalytic cleft. 3A2 variants with high potency, selectivity, and improved affinity and proteolytic stability were isolated from a random mutagenesis library...
August 13, 2018: Biotechnology and Bioengineering
Nicole K Paulk, Katja Pekrun, Gregory W Charville, Katie Maguire-Nguyen, Michael N Wosczyna, Jianpeng Xu, Yue Zhang, Leszek Lisowski, Bryan Yoo, Jose G Vilches-Moure, Gordon K Lee, Joseph B Shrager, Thomas A Rando, Mark A Kay
Skeletal muscle is ideal for passive vaccine administration as it is easily accessible by intramuscular injection. Recombinant adeno-associated virus (rAAV) vectors are in consideration for passive vaccination clinical trials for HIV and influenza. However, greater human skeletal muscle transduction is needed for therapeutic efficacy than is possible with existing serotypes. To bioengineer capsids with therapeutic levels of transduction, we utilized a directed evolution approach to screen libraries of shuffled AAV capsids in pools of surgically resected human skeletal muscle cells from five patients...
September 21, 2018: Molecular Therapy. Methods & Clinical Development
Camilo E Quevedo, Abimael Cruz-Migoni, Nicolas Bery, Ami Miller, Tomoyuki Tanaka, Donna Petch, Carole J R Bataille, Lydia Y W Lee, Phillip S Fallon, Hanna Tulmin, Matthias T Ehebauer, Narcis Fernandez-Fuentes, Angela J Russell, Stephen B Carr, Simon E V Phillips, Terence H Rabbitts
Targeting specific protein-protein interactions (PPIs) is an attractive concept for drug development, but hard to implement since intracellular antibodies do not penetrate cells and most small-molecule drugs are considered unsuitable for PPI inhibition. A potential solution to these problems is to select intracellular antibody fragments to block PPIs, use these antibody fragments for target validation in disease models and finally derive small molecules overlapping the antibody-binding site. Here, we explore this strategy using an anti-mutant RAS antibody fragment as a competitor in a small-molecule library screen for identifying RAS-binding compounds...
August 9, 2018: Nature Communications
Bronwyn M Gunn, Wen-Han Yu, Marcus M Karim, Jennifer M Brannan, Andrew S Herbert, Anna Z Wec, Peter J Halfmann, Marnie L Fusco, Sharon L Schendel, Karthik Gangavarapu, Tyler Krause, Xiangguo Qiu, Shinhua He, Jishnu Das, Todd J Suscovich, Jonathan Lai, Kartik Chandran, Larry Zeitlin, James E Crowe, Douglas Lauffenburger, Yoshihiro Kawaoka, Gary P Kobinger, Kristian G Andersen, John M Dye, Erica Ollmann Saphire, Galit Alter
The recent Ebola virus (EBOV) epidemic highlighted the need for effective vaccines and therapeutics to limit and prevent outbreaks. Host antibodies against EBOV are critical for controlling disease, and recombinant monoclonal antibodies (mAbs) can protect from infection. However, antibodies mediate an array of antiviral functions including neutralization as well as engagement of Fc-domain receptors on immune cells, resulting in phagocytosis or NK cell-mediated killing of infected cells. Thus, to understand the antibody features mediating EBOV protection, we examined specific Fc features associated with protection using a library of EBOV-specific mAbs...
August 8, 2018: Cell Host & Microbe
Rachel Ryu, Kristina E Ward
Purpose: Atezolizumab is a programmed death ligand 1 (PDL-1) blocking antibody that was approved for metastatic non-small cell lung cancer (NSCLC) in patients with disease progression. Various studies have been initiated to explore the effectiveness of atezolizumab among different patient cohorts and disease statuses, including as first-line therapy. The purpose of this paper is to identify and summarize the trials that use atezolizumab as a first-line agent in chemotherapy-naïve patients with NSCLC. Methods: A database search was performed on Pubmed, Embase, and Wiley Cochrane Library-Central Register of Controlled Trials to identify clinical trials using atezolizumab as first-line therapy in NSCLC...
2018: Frontiers in Oncology
Hao Qu, Lu Wang, Jian Liu, Lei Zheng
Cytometric bead assays have caught much attention because of their many exceptional advantages. Unfortunately, the immobilization of existing molecular recognition elements including monoclonal antibodies and aptamers onto solid particles may lead to the function failure of the molecular recognition elements since they are generally obtained in free state. Herein we develop a powerful screening approach for direct and rapid discovery of aptamer based cytometric bead assays (AB-CBAs) by individually measuring the functional activity of every aptamer particles in a library and sorting them at rates of up to 108 particles per hour...
August 7, 2018: ACS Sensors
Veeramohan Veerapandian, Jan Ole Ackermann, Yogesh Srivastava, Vikas Malik, Mingxi Weng, Xiaoxiao Yang, Ralf Jauch
Directed biomolecular evolution is widely used to tailor and enhance enzymes, fluorescent proteins, and antibodies but has hitherto not been applied in the reprogramming of mammalian cells. Here, we describe a method termed directed evolution of reprogramming factors by cell selection and sequencing (DERBY-seq) to identify artificially enhanced and evolved reprogramming transcription factors. DERBY-seq entails pooled screens with libraries of positionally randomised genes, cell selection based on phenotypic readouts, and genotyping by amplicon sequencing for candidate identification...
July 27, 2018: Stem Cell Reports
Christian Schröter, Jan Beck, Simon Krah, Stefan Zielonka, Achim Doerner, Laura Rhiel, Ralf Günther, Lars Toleikis, Harald Kolmar, Björn Hock, Stefan Becker
In this study, we present a multiparameter screening procedure for the identification of target-specific antibodies with prescribed properties. Based on B cell receptor gene repertoires from transgenic rats, yeast surface display libraries were generated, and high-affinity human antibodies were readily isolated. We demonstrate that specific desirable features, i.e., species' cross-reactivity and a broad epitope coverage can be integrated into the screening procedure using high-throughput fluorescence-activated cell sorting...
August 3, 2018: Molecular Biotechnology
Aaron L Nichols, Kaori Noridomi, Chris R Hughes, Farzad Jalali-Yazdi, J Brek Eaton, Lan Huong Lai, Gaurav Advani, Ronald J Lukas, Henry A Lester, Lin Chen, Richard W Roberts
The nicotinic acetylcholine receptors are pentameric ligand-gated ion channels that play a central role in neuronal and neuromuscular signal transduction. Here, we have developed FANG ligands - Fibronectin Antibody-mimetic Nicotinic acetylcholine receptor Generated ligands - using mRNA display. We generated a trillion-member primary e10FnIII library to target a stabilized α1 nicotinic subunit (α211). This library yielded 270,000 independent potential protein-binding ligands. The lead sequence, α1-FANG1, represented 25% of all library sequences, showed the highest-affinity binding, and competed with α-bungarotoxin (α-Btx)...
July 30, 2018: ACS Chemical Biology
Mehreen Zaka, Bilal Haider Abbasi, Serdar Durdagi
Tumor necrosis factor alpha (TNF-α) is a multifunctional cytokine that acts as a central biological mediator for critical immune functions, including inflammation, infection, and antitumor responses. It plays pivotal role in auto-immune diseases like rheumatoid arthritis (RA). The synthetic antibodies etanercept, infliximab, and adalimumab are approved drugs for the treatment of inflammatory diseases bind to TNF-α directly, preventing its association with the tumor necrosis factor receptor (TNFR). These biologics causes serious side effects such as triggering an autoimmune anti-antibody response or the weakening of the body's immune defenses...
July 26, 2018: Journal of Biomolecular Structure & Dynamics
Wenshuai Zhang, Xiaoyan Zeng, Ying Chi, Jingxian Liu, Yongjun Jiao
Objective To construct a human phage display library against severe fever with thrombocytopenia syndrome (SFTS) virus. Methods The total RNA was isolated from peripheral blood lymphocytes of 8 patients with SFTS and cDNA was amplified. The genes of light-chain (Vκ and Vλ) and heavy-chain (VH) were amplified by PCR. The scFv gene was linked by overlap-PCR and cloned into the vector pComb3XSS, and then transformed into XL1-Blue cells for the phage antibody library construction. After detecting the recombinant rate and the library repertoire, we screened the antibodies against SFTS virus by biopanning with immobilized virus antigen...
May 2018: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Harvinder Talwar, Samer Najeeb Hanoudi, Sorin Draghici, Lobelia Samavati
Tuberculosis (TB) is caused by Mycobacterium tuberculosis (MTB) and transmitted through inhalation of aerosolized droplets. Eighty-five percent of new TB cases occur in resource-limited countries in Asia and Africa and fewer than 40% of TB cases are diagnosed due to the lack of accurate and easy-to-use diagnostic assays. Currently, diagnosis relies on the demonstration of the bacterium in clinical specimens by serial sputum smear microscopy and culture. These methods lack sensitivity, are time consuming, expensive, and require trained personnel...
July 19, 2018: Viruses
Nika Kruljec, Peter Molek, Vesna Hodnik, Gregor Anderluh, Tomaž Bratkovič
Affinity chromatography based on bacterial immunoglobulin (Ig)-binding proteins represents the cornerstone of therapeutic antibody downstream processing. However, there is a pressing need for more robust affinity ligands that would withstand the harsh column sanitization conditions, while still displaying high selectivity for antibodies. Here, we report the development of linear peptide IgG ligands, identified from combinatorial phage-display library screens. The lead peptide was shown to compete with staphylococcal protein A for the IgG Fc region...
August 1, 2018: Bioconjugate Chemistry
Shoichi Naito, Tatsuya Takahashi, Junji Onoda, Shoko Uemura, Naoki Ohyabu, Hiroshi Takemoto, Shoji Yamane, Ikuo Fujii, Shin-Ichiro Nishimura, Yoshito Numata
Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O -glycan core structures have already been developed. However, most of them show low specificities toward O -glycan structures and/or low affinity toward a monovalent epitope. In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines...
November 30, 2017: ACS Omega
Emily G McDonald, Lyne Bissonette, Stephanie Ensworth, Natalie Dayan, Ann E Clarke, Stephanie Keeling, Sasha Bernatsky, Evelyne Vinet
OBJECTIVE: Few data exist to guide the frequency and type of monitoring in systemic lupus erythematosus (SLE) pregnancies. A systematic literature review was performed to address this gap in the literature. METHODS: A systematic review of original articles (1975-2015) was performed using Medline, Embase, and Cochrane Library. We included search terms for SLE, pregnancy, and monitoring. We also hand-searched reference lists, review articles, and grey literature for additional relevant articles...
July 15, 2018: Journal of Rheumatology
Sylvia Annabel Dass, Mohd Nor Norazmi, Armando Acosta Dominguez, Maria Elena Sarmiento Garcia San Miguel, Gee Jun Tye
The discovery of heat shock protein 16 kDa antigen protein has deepen the understanding of latent tuberculosis since it was found to be primarily expressed by Mycobacterium tuberculosis during latent phase leading to the rapid optimization and development in terms of diagnosis and therapeutics. Recently, T cell receptor-like antibody has been explored extensively targeting various diseases due to its dual functionality (T cell receptor and antibody). In this study, a TCR-like domain antibody (A2/Ab) with the binding capacity to Mtb heat shock protein (HSP) 16 kDa antigen presented by major histocompatible complex (MHC) HLA-A*02 was successfully generated via biopanning against human domain antibody library...
July 4, 2018: Molecular Immunology
Junko Hamamoto, Hiroyuki Yasuda, Yosuke Nonaka, Masatoshi Fujiwara, Yoshikazu Nakamura, Kenzo Soejima, Tomoko Betsuyaku
Cancers, including lung cancer, are a leading cause of death worldwide. To overcome this deadly disease, multiple modality inhibitors have been developed. These include cytotoxic agents, molecular targeted small molecules, such as tyrosine kinase inhibitors, and neutralizing antibodies. An aptamer is a short single-stranded nucleic acid molecule that is selected in vitro from a large random sequence library based on its high and specific affinity to a target molecule. Aptamers can be applied to therapeutics of various types of diseases, including cancer, due to their strong and specific neutralizing activities...
July 10, 2018: Biochemical and Biophysical Research Communications
Yurika Yada, Melbourne Rio Talactac, Kodai Kusakisako, Emmanuel Pacia Hernandez, Remil Linggatong Galay, Masako Andoh, Kozo Fujisaki, Tetsuya Tanaka
Ticks are key vectors of some important diseases of humans and animals. Although they are carriers of disease agents, the viability and development of ticks are not harmed by the infectious agents due to their innate immunity. Antimicrobial peptides directly protect hosts against pathogenic agents such as viruses, bacteria, and parasites. Among the identified and characterized antimicrobial peptides, defensins have been considerably well studied. Defensins are commonly found among fungi, plants, invertebrates, and vertebrates...
July 2018: Journal of Invertebrate Pathology
Barry W Neun, Yechezkel Barenholz, Janos Szebeni, Marina A Dobrovolskaia
Infusion reactions (IRs) are common immune-mediated side effects in patients treated with a variety of drug products, including, but not limited to, nanotechnology formulations. The mechanism of IRs is not fully understood. One of the best studied mechanisms of IRs to nanomedicines is the complement activation. However, it is largely unknown why some patients develop reactions to nanomedicines while others do not, and why some nanoparticles are more reactogenic than others. One of the theories is that the pre-existing anti-polyethylene glycol (PEG) antibodies initiate the complement activation and IRs in patients...
July 12, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
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