Review
Microbiology
Ana S. S. Andre, Isa Moutinho, Joana N. R. Dias, Frederico Aires-da-Silva
Summary: The discovery of hybridoma technology and the development of phage display technology have opened a new era in antibody research and clinical development. Over the past thirty years, display selection technologies such as phage display have been widely promoted as ideal alternatives to traditional methods, with significant advancements in design, construction, and selection of antibody libraries. With the emergence of new therapeutic antibodies, phage display is expected to continue playing a central role, especially in challenging cases. In vivo phage display methods offer an advantage over conventional in vitro procedures for generating therapeutic antibodies against difficult targets and unmet medical needs.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biochemical Research Methods
Charlotte Rimbault, Pelle D. Knudsen, Anna Damsbo, Kim Boddum, Hanif Ali, Celeste M. Hackney, Lars Ellgaard, Markus -Frederik Bohn, Andreas H. Laustsen
Summary: Phage display technology is a powerful tool for selecting monoclonal antibodies against a diverse set of antigens. However, generating monoclonal antibodies against animal toxins is challenging due to difficulties in obtaining them from venom. This study successfully expressed alpha-cobratoxin in E. coli and used it as an antigen in a phage display selection campaign, resulting in the isolation of a scFv with similar binding affinity to a control scFv against the native toxin. The selected scFv inhibits the interaction between the acetylcholine receptor and the native toxin in vitro, representing the first entirely in vitro antibody selection strategy for generating neutralizing monoclonal antibodies against a snake toxin.
Article
Pharmacology & Pharmacy
Sandra Isabel Aguiar, Joana N. R. Dias, Ana Santos Andre, Marta Lisete Silva, Diana Martins, Belmira Carrapico, Miguel Castanho, Joao Carrico, Marco Cavaco, Maria Manuela Gaspar, Rui Jorge Nobre, Luis Pereira de Almeida, Soraia Oliveira, Lurdes Gano, Joao D. G. Correia, Carlos Barbas, Joao Goncalves, Vera Neves, Frederico Aires-da-Silva
Summary: The major bottleneck in CNS drug development is the discovery of molecules that can cross BBB. Using nano-delivery strategies and in vivo cell immunization to construct sdAb library targeting BBB endothelial cell receptors, specific nano-antibody scaffolds with promising properties for brain targeting and drug delivery were successfully selected.
Article
Cell Biology
Federico Bertoglio, Viola Fuehner, Maximilian Ruschig, Philip Alexander Heine, Leila Abassi, Thomas Kluenemann, Ulfert Rand, Doris Meier, Nora Langreder, Stephan Steinke, Rico Ballmann, Kai-Thomas Schneider, Kristian Daniel Ralph Roth, Philipp Kuhn, Peggy Riese, Dorina Schaeckermann, Janin Korn, Allan Koch, M. Zeeshan Chaudhry, Kathrin Eschke, Yeonsu Kim, Susanne Zock-Emmenthal, Marlies Becker, Margitta Scholz, Gustavo Marcal Schmidt Garcia Moreira, Esther Veronika Wenzel, Giulio Russo, Hendrikus S. P. Garritsen, Sebastian Casu, Andreas Gerstner, Guenter Roth, Julia Adler, Jakob Trimpert, Andreas Hermann, Thomas Schirrmann, Stefan Duebel, Andre Frenzel, Joop Van den Heuvel, Luka Cicin-Sain, Maren Schubert, Michael Hust
Summary: Researchers have developed a human neutralizing antibody against SARS-CoV-2, which shows promising efficacy in animal models and is currently undergoing clinical trials for the treatment of moderate to severe COVID-19. The antibody binds to the receptor-binding domain (RBD) of the spike protein to inhibit the virus, and is not affected by many known RBD mutations.
Article
Chemistry, Multidisciplinary
Maximilian Brueckner, Johanna Simon, Katharina Landfester, Volker Mailaender
Summary: Antibody-modified drug delivery systems in the nano-range show potential in overcoming current challenges for disease treatment due to their high specificity, but no clinically approved antibody-bound nanocarrier exists. Different conjugation strategies influence the spatial orientation of antibodies on the nanocarriers' surface, impacting the success of antibody functionalized drug delivery systems. In this study, two conjugation strategies were compared, showing that both thiol-maleimide coupling and copper-free click chemistry can induce dendritic cell uptake, with orientation of antibodies affecting the targeting efficiency.
Article
Biology
Fanlin Li, Wei Xu, Xiaoqing Zhang, Wanting Wang, Shan Su, Ping Han, Haiyong Wang, Yanqin Xu, Min Li, Lilv Fan, Huihui Zhang, Qiang Dai, Hao Lin, Xinyue Qi, Jie Liang, Xin Wang, Shibo Jiang, Youhua Xie, Lu Lu, Xuanming Yang
Summary: Neutralizing antibodies are not as effective in therapeutic models as in prophylactic models due to limited efficacy in eliminating virus-producing cells. A SARS-CoV-2 spike-targeting bispecific T-cell engager (S-BiTE) strategy is presented, which blocks viral entry and eliminates infected cells through T cell-mediated cytotoxicity. S-BiTE is effective against both original and Delta variants of SARS-CoV-2, suggesting potential application against immune-escaping variants. In a humanized mouse model with live SARS-CoV-2 infection, S-BiTE significantly reduces viral load compared to neutralization treatment only.
COMMUNICATIONS BIOLOGY
(2023)
Review
Biochemistry & Molecular Biology
Longguang Jiang, Cai Yuan, Mingdong Huang
Summary: Serine proteases are a large family of enzymes critical for physiological processes, with the autolysis loop on these enzymes showing high variability and potential for regulating catalytic activity and specificity of inhibitors. The autolysis loop appears to be a sensitive allosteric site that can be targeted to develop pharmacological agents for interventions in activities such as blood coagulation.
Article
Microbiology
Jana Hruskovicova, Katarina Bhide, Patricia Petrouskova, Zuzana Tkacova, Evelina Mochnacova, Jan Curlik, Mangesh Bhide, Amod Kulkarni
Summary: In this study, single-domain antibodies (sdAbs) were developed against the domain III (DIII) of WNV's envelope glycoprotein to interrupt the interaction between DIII and the human brain microvascular endothelial cells (hBMEC). Among the sdAbs, sdAb(A10) showed optimum stability, neutralized the WNV-like particles, and had low hemolytic activity and cytotoxicity. Therefore, sdAb(A10) is considered a specific therapy for WNV-induced neuropathogenesis.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biochemical Research Methods
Pedro G. M. Condelipes, Pedro Mendes Fontes, Ana Godinho-Santos, Eduardo J. S. Bras, Vanda Marques, Marta B. B. Afonso, Cecilia M. P. Rodrigues, Virginia Chu, Joao Goncalves, Joao Pedro Conde
Summary: Antibody therapy has played a significant role in the treatment of various diseases, including cancer. Phage display technology has emerged as a promising approach to expedite the development of antibody therapies. Microfluidics, a technology that replicates physiologically relevant conditions, can enhance screening efficacy. In this study, a polydimethylsiloxane/polystyrene-based microfluidic system was used to continuously perfuse cancer cells. Through this system, the binding kinetics and specificity of engineered phages targeting the CXCR4 receptor were investigated. The results demonstrated the simplicity and ease of use of the microfluidic device design, which could serve as a generic platform for studying and optimizing therapies based on interaction with biological entities.
Review
Pharmacology & Pharmacy
Xue-Kai Wang, Zong-Gen Peng
Summary: NAFLD and its advanced stage NASH pose a global public health threat, with no specific drugs approved for NASH treatment yet promising candidates in development. LSECs have been shown to play a crucial role in liver inflammation of NAFLD, offering potential as a therapeutic target for controlling liver inflammation.
FRONTIERS IN PHARMACOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Frederick R. Cross, Peter C. Fridy, Natalia E. Ketaren, Fred D. Mast, Song Li, J. Paul Olivier, Kresti Pecani, Brian T. Chait, John D. Aitchison, Michael P. Rout
Summary: COVID-19, caused by the SARS-CoV-2 coronavirus, is a global health issue with emerging variants. Nanobodies derived from camels show potential to neutralize the virus. By optimizing a yeast display method, a large repertoire of high-affinity and neutralizing nanobodies against different SARS-CoV-2 variants was identified. The yeast display approach is comparable to mass spectrometry and allows for parallel interrogation of a vast number of nanobodies, maximizing the likelihood of discovering broadly specific reagents.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2023)
Article
Multidisciplinary Sciences
Shutong Qian, Jiayi Mao, Qiuyu Zhao, Binfan Zhao, Zhimo Liu, Bolun Lu, Liucheng Zhang, Xiyuan Mao, Yuguang Zhang, Danru Wang, Xiaoming Sun, Wenguo Cui
Summary: In this study, CX3CL1-functionalized apoptotic body nanovesicles (nABs) were used to target endothelial cells (ECs) and promote angiogenesis in the chronic wound process. The Find-eat strategy was implemented to amplify the Find-eat signal and stimulate ECs in the hypoxic microenvironment. In vitro and in vivo experiments demonstrated that nABs had good biocompatibility, promoted wound closure, and released angiogenic drugs for new blood vessel formation in diabetic wounds. These receptor-functionalized nABs provide a novel strategy for chronic diabetic wound healing.
Article
Multidisciplinary Sciences
Briana Martinez, Gergey Alzaem Mousa, Kiera Fleck, Tara MacCulloch, Chris W. Diehnelt, Nicholas Stephanopoulos, Sarah E. Stabenfeldt
Summary: The heterogeneous pathophysiology of traumatic brain injury poses challenges for diagnostics and therapeutics. This study used a unique discovery pipeline to identify novel targeting motifs that recognize specific temporal phases of TBI pathology. These findings provide a foundation for the development of targeted TBI therapeutics and diagnostics.
Article
Computer Science, Information Systems
Yuzhe Li, Yong Liu, Bo Li, Weiping Wang, Nan Liu
Summary: As privacy-preserving data analysis becomes increasingly important, the trade-off between privacy guarantee and analysis accuracy has been a concern in practice. This paper focuses on private ERM and explores the effect of the differential privacy parameter c on the utility of the learning model. The proposed approach provides a practical way to estimate utility under different privacy requirements. The high estimation accuracy and broad applicability make it valuable in practical applications.
COMPUTERS & SECURITY
(2023)
Review
Biochemistry & Molecular Biology
Catarina J. G. Pinto, Maria Angeles Avila-Galvez, Yilong Lian, Pedro Moura-Alves, Claudia Nunes dos Santos
Summary: The Aryl Hydrocarbon Receptor (AHR) is a ligand-dependent transcription factor that can control complex transcriptional processes in various cell types and has been associated with diseases such as inflammatory bowel diseases (IBD). Different compounds, including xenobiotics, natural compounds, and host-derived metabolites, have been identified as ligands of the AHR. Dietary (poly)phenols, which have been extensively studied for their pleiotropic activities, may also modulate the AHR. However, the extensive metabolism of dietary (poly)phenols in the gut, particularly by the gut microbiota, may result in the production of gut phenolic metabolites that could play a key role in modulating AHR and influencing inflammatory processes in the gut.
Review
Engineering, Chemical
Yan Zhang, Bekir Engin Eser, Peter Kristensen, Zheng Guo
CHINESE JOURNAL OF CHEMICAL ENGINEERING
(2020)
Article
Chemistry, Multidisciplinary
Maja Thim Larsen, Ole A. Mandrup, Karen Kraemmer Schelde, Yonglun Luo, Karina Dalsgaard Sorensen, Frederik Dagnaes-Hansen, Jason Cameron, Magnus Stougaard, Torben Steiniche, Kenneth A. Howard
JOURNAL OF CONTROLLED RELEASE
(2020)
Article
Biology
Ole A. Mandrup, Sui Ching Ong, Simon Lykkemark, Anders Dinesen, Imke Rudnik-Jansen, Niels Frederik Dagnaes-Hansen, Jan Terje Andersen, Luis Alvarez-Vallina, Kenneth A. Howard
Summary: This study introduces a programmable serum half-life extension platform based on human albumin sequences fused with bispecific T-cell engagers, offering potential long-lasting effects, better patient compliance, and the ability to tailor pharmacokinetics to maximize therapeutic efficacy and safety of immuno-oncology targeted biologics.
COMMUNICATIONS BIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Yan Zhang, Bekir Engin Eser, Georgios Kougioumtzoglou, Zekiye Eser, Michal Poborsky, Shigenobu Kishino, Michiki Takeuchi, Jun Ogawa, Peter Kristensen, Zheng Guo
Summary: This research investigates the regioselectivity of fatty acid hydratase-mediated hydroxy fatty acid production, achieving a broader conversion of substrates through amino acid mutations and residue variations, further optimizing the high-value products.
BIOCHEMICAL ENGINEERING JOURNAL
(2021)
Article
Biochemistry & Molecular Biology
Yan Zhang, Niels Mikkel Dyrby Breum, Sune Schubert, Negin Hashemi, Rikke Kyhnau, Marius Sandholt Knauf, Masuthan Mathialakan, Michiki Takeuchi, Shigenobu Kishino, Jun Ogawa, Peter Kristensen, Zheng Guo, Bekir Engin Eser
Summary: Research on semi-rational protein engineering to alter the binding site residues of Lactobacillus Acidophilus' fatty acid hydratase-1 for improved regioselectivity; although not achieving a completely new regioselectivity, a shift towards cis-5, cis-8, cis-11, cis-14, cis-17-eicosapentaenoic acid was observed; mutants like S393/S395 showed excellent regioselectivity, producing a single hydroxy fatty acid product, which is advantageous from an application standpoint.
Article
Chemistry, Multidisciplinary
Wei Fan, Tao Du, Aida Droce, Lars R. Jensen, Randall E. Youngman, Xiangting Ren, Leonid Gurevich, Mathieu Bauchy, Peter Kristensen, Bengang Xing, Donghong Yu, Morten M. Smedskjaer
Summary: This study investigates the deformation and fracture behavior of inorganic-polymer hybrids and finds that the noncovalent intermolecular interactions between polymer and silica networks play a crucial role. The study also explores three methods to improve the balance between strength and toughness of the hybrids and demonstrates the optimized mechanical properties and biocompatibility of the hybrid material.
Article
Biochemistry & Molecular Biology
Josephine Geertsen Keller, Kirstine Mejlstrup Hymoller, Maria Eriksen Thorsager, Noriko Y. Hansen, Jens Uldum Erlandsen, Cinzia Tesauro, Anne Katrine W. Simonsen, Anne Bech Andersen, Kamilla Vands-Petersen, Lise Lolle Holm, Magnus Stougaard, Brage Storstein Andresen, Peter Kristensen, Rikke Frohlich, Birgitta R. Knudsen
Summary: The function of human topoisomerase 1 (TOP1) in regulating G-quadruplex (G4) formation in the MYC P1 promoter has been investigated, revealing its involvement in the regulation of G4 formation and its potential as a regulator in gene expression.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Engineering, Biomedical
Elisabeth Fuchs, Imke Rudnik-Jansen, Anders Dinesen, Denis Selnihhin, Ole Aalund Mandrup, Kader Thiam, Jorgen Kjems, Finn Skou Pedersen, Kenneth A. Howard
Summary: The emergence of new SARS-CoV-2 mutants and breakthrough infections has led to the need for antiviral therapeutics. This study presents a recombinant human albumin ACE2 genetic fusion that can increase the plasma half-life and inhibit virus entry into cells. The fusion protein is shown to bind to the SARS-CoV-2 spike protein and inhibit viral infection.
ACTA BIOMATERIALIA
(2022)
Article
Chemistry, Analytical
Andreas Madsen, Oscar Mejias-Gomez, Lasse E. Pedersen, Kerstin Skovgaard, Peter Kristensen, Steffen Goletz
Summary: Simultaneous targeting of different antigens with bispecific antibodies (bsAbs) has high therapeutic potential but presents challenges in analysis. Flow-induced dispersion analysis (FIDA) allows in-depth characterization of bsAbs under near-native conditions, determining equilibrium dissociation constants and analyzing binding cooperativity. FIDA is a versatile tool with low material consumption and high matrix tolerability for format selection and characterization of complex protein therapeutics.
ANALYTICAL CHEMISTRY
(2022)
Article
Medicine, Research & Experimental
Andreas V. Madsen, Peter Kristensen, Alexander K. Buell, Steffen Goletz
Summary: This study demonstrates a simple and efficient method to generate high-quality bsAbs with both binding functionalities intact by fusing sdAbs onto IgG scaffolds through flexible linkers. By systematically comparing sdAb fusion strategies, the results show that fusion of sdAbs to the heavy chain promotes good expression, stability, and efficient binding to both antigens. The study also provides a toolbox of complementary methods for in-depth analysis of key features, such as dual antigen binding and stability, to ensure high bsAb quality.
Article
Chemistry, Analytical
Jinhua Dong, Bhagat Banwait, Hiroshi Ueda, Peter Kristensen
Summary: Quenchbodies, a type of biosensor, use antibodies labeled with a fluorescent dye to detect proteins. The dye is quenched in the absence of an antigen, but de-quenched in its presence. This study demonstrates the development of mini Q-bodies using a single-domain heavy chain V-H antibody and different dyes. The results confirm the applicability of this method in detecting proteins.
Article
Medicine, Research & Experimental
Oscar Mejias-Gomez, Andreas V. Madsen, Kerstin Skovgaard, Lasse E. Pedersen, J. Preben Morth, Timothy P. Jenkins, Peter Kristensen, Steffen Goletz
Summary: This study analyzes human antibody sequences to understand the factors determining antibody diversity and specificity. The study reveals the importance of specific residue positions and their influence on antibody characteristics. These findings have significant implications for the development and optimization of antibody therapeutics.
Article
Biochemical Research Methods
Oscar Mejias-Gomez, Andreas V. Madsen, Lasse E. Pedersen, Peter Kristensen, Steffen Goletz
Summary: This study presents a system based on split beta-lactamase complementation, which can remove OFF-frame clones and increase functional diversity more quickly and efficiently, suitable for constructing randomized libraries. The system can eliminate OFF-frame clones containing stop codons or frameshifts in starting mixtures, enriching the IN-frame clones to about 70%.
Meeting Abstract
Chemistry, Applied
Yan Zhang, Bekir E. Eser, Zheng Guo
JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY
(2020)
Article
Chemistry, Analytical
Andreas Madsen, Oscar Mejias-Gomez, Lasse E. Pedersen, Kerstin Skovgaard, Peter Kristensen, Steffen Goletz
Summary: FIDA technology enables in-depth characterization of bsAbs with diverse molecular architectures and valencies, analyzing their bispecific binding functionality without being affected by molecular format, making it a versatile tool for selecting and characterizing complex protein therapeutics.
ANALYTICAL CHEMISTRY
(2022)