A method to determine the mode of binding for GCPII inhibitors using bio-layer interferometry

Choy, Cindy J.; Berkman, Clifford E.

doi:10.3109/14756366.2015.1132208

Cited by 5 publications

(2 citation statements)

References 16 publications

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“…One milligram of human LOX-1 protein (Sino Biological Inc., China) was biotinylated at a 1:1 ratio with EZLink Sulfo-NHS-Biotin (Pierce), so that it would bind to the SA biosensor. The assays were measured in a 96-well format according to a method from a previous study [18]. Kinetic responses were fitted to a 1-site binding model to obtain the values for the kinetic association (k…”

Section: Affinity Analysis By Biolayer Interferometry (Bli)mentioning

confidence: 99%

Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 in Escherichia coli and Brevibacillus choshinensis

Xiang

Kong

et al. 2017

Journal of Microbiology and Biotechnology

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The single-chain variable fragment (scFv) against lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a promising molecule for its potential use in the diagnosis and immunotherapy of atherosclerosis. Producing this scFv in several milligram amounts could be the starting point for further engineering and application of the scFv. In this study, the abundant expression of the anti-LOX-1 scFv was attempted using () and . The scFv had limited soluble yield in, but it was efficiently secreted by . The optimized fermentation was determined using the Plackett-Burman screening design and response surface methodology, under which the yield reached up to 1.5 g/l in a 5-L fermentor. Moreover, the properties of the scFvs obtained from the two expression systems were different. The antigen affinity, transition temperature, and particle diameter size were 1.01E-07 M, 55.2 ± 0.3°C, and 9.388 nm for the scFv expressed by, and 4.53E-07 M, 52.5 ± 0.3°C, and 13.54 nm for the scFv expressed by . This study established an efficient scale-up production methodology for the anti-LOX-1 scFv, which will boost its use in LOX-1-based therapy.

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Section: Affinity Analysis By Biolayer Interferometry (Bli)mentioning

confidence: 99%

Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 in Escherichia coli and Brevibacillus choshinensis

Xiang

Kong

et al. 2017

Journal of Microbiology and Biotechnology

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“…Table S4 shows several major cases of the application of BLI biosensors for drug screening. The drug targets cover glutamate carboxypeptidase II, 28 glutaminase, 29 double-stranded DNA, 30 and protein−protein interactions, such as tissue transglutaminase and fibronectin interaction, 31 and SARSCoV-2 RBD and hACE2 interaction. 32 Advantages of Biolayer Interferometry Biosensors.…”

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confidence: 99%

Tailored Biosensors for Drug Screening, Efficacy Assessment, and Toxicity Evaluation

et al. 2021

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Biosensors have been flourishing in the field of drug discovery with pronounced developments in the past few years. They facilitate the screening and discovery of innovative drugs. However, there is still a lack of critical reviews that compare the merits and shortcomings of these biosensors from a pharmaceutical point of view. This contribution presents a critical and up-to-date overview on the recent progress of tailored biosensors, including surface plasmon resonance, fluorescent, photoelectrochemical, and electrochemical systems with emphasis on their mechanisms and applications in drug screening, efficacy assessment, and toxicity evaluation. Multiple functional nanomaterials have also been incorporated into the biosensors. Representative examples of each type of biosensors are discussed in terms of design strategy, response mechanism, and potential applications. In the end, we also compare the results and summarize the major insights gained from the works, demonstrating the challenges and prospects of biosensors-assisted drug discovery.

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Enhanced Bioactivity of the Anti-LOX-1 scFv Engineered by Multimerization Strategy

Xie

Liu

2017

Appl Biochem Biotechnol

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Single-chain variable fragment (scFv) antibodies as therapeutic agents have the potential to reduce the production cost and immunogenicity relative to monoclonal antibodies, but their monovalency and lack of a fragment crystallizable region can lead to reduced function. Multimerization is one strategy for recovering the function; however, their application is limited by the production of multimeric proteins. In our previous study, an anti-lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) scFv showed potential use in diagnosis and therapy of atherosclerotic diseases, but is limited by its inherent low antigen-binding activity. In this study, to improve the efficacy of the anti-LOX-1 scFv, we constructed the anti-LOX-1 scFv multimers by modifying the linker length between the variable domains of the scFv or by fusing the scFv with self-merization domains and expressed these scFv multimers in Brevibacillus choshinensis hosts. After optimization, all of the scFv multimers obtained efficient secretion expression. Compared with the scFv monomer, the multimers that are successfully fractionated displayed increased neutralization activity and showed elevated antigen-binding avidity, especially the tetramer, which improved the antigen avidity by two orders of magnitude. Moreover, the scFv dimer and the tetramer both displayed better stability and longer half-life in serum, which can be attractive candidates for the next-generation anti-LOX-1 therapeutic antibody.

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A method to determine the mode of binding for GCPII inhibitors using bio-layer interferometry

Cited by 5 publications

References 16 publications

Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 in Escherichia coli and Brevibacillus choshinensis

Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 in Escherichia coli and Brevibacillus choshinensis

Tailored Biosensors for Drug Screening, Efficacy Assessment, and Toxicity Evaluation

Enhanced Bioactivity of the Anti-LOX-1 scFv Engineered by Multimerization Strategy

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