scholarly journals Experimental Validation of a Fragment Library for Lead Discovery Using SPR Biosensor Technology

2010 ◽  
Vol 16 (1) ◽  
pp. 15-25 ◽  
Author(s):  
Malin Elinder ◽  
Matthis Geitmann ◽  
Thomas Gossas ◽  
Per Källblad ◽  
Johan Winquist ◽  
...  
Keyword(s):  
Surface Plasmon Resonance ◽  
Human Serum Albumin ◽  
Drug Targets ◽  
Experimental Validation ◽  
High Sensitivity ◽  
Lead Discovery ◽  
Experimental Procedure ◽  
Spr Biosensor ◽  
Fragment Library ◽  
Hiv 1

A new fragment library for lead discovery has been designed and experimentally validated for use in surface plasmon resonance (SPR) biosensor-based screening. The 930 compounds in the library were selected from 4.6 million commercially available compounds using a series of physicochemical and medicinal chemistry filters. They were screened against 3 prototypical drug targets: HIV-1 protease, thrombin and carbonic anhydrase, and a nontarget: human serum albumin. Compound solubility was not a problem under the conditions used for screening. The high sensitivity of the sensor surfaces allowed the detection of interactions for 35% to 97% of the fragments, depending on the target protein. None of the fragments was promiscuous (i.e., interacted with a stoichiometry ≥5:1 with all 4 proteins), and only 2 compounds dissociated slowly from all 4 proteins. The use of several targets proved valuable since several compounds would have been disqualified from the library on the grounds of promiscuity if fewer target proteins had been used. The experimental procedure allowed an efficient evaluation and exploration of the new fragment library and confirmed that the new library is suitable for SPR biosensor-based screening.

scholarly journals Multiplexed experimental strategies for fragment library screening using SPR biosensors

2020 ◽  
Author(s):  
Edward A. FitzGerald ◽  
Darius Vagrys ◽  
Giulia Opassi ◽  
Hanna F. Klein ◽  
David J. Hamilton ◽  
...  
Keyword(s):  
Library Screening ◽  
Lead Discovery ◽  
Chemical Library ◽  
Innovative Strategies ◽  
Spr Biosensor ◽  
Screening Experiments ◽  
Fragment Library ◽  
Experimental Approaches ◽  
Orthogonal Methods ◽  
Experimental Strategies

AbstractSurface plasmon resonance biosensor technology (SPR) is ideally suited for fragment-based lead discovery. However, generally suitable experimental procedures or detailed protocols are lacking, especially for structurally or physico-chemically challenging targets or when tool compounds are lacking. Success depends on accounting for the features of both the target and the chemical library, purposely designing screening experiments for identification and validation of hits with desired specificity and mode-of-action, and availability of orthogonal methods capable of confirming fragment hits. By adopting a multiplexed strategy, the range of targets and libraries amenable to an SPR biosensor-based approach for identifying hits is considerably expanded. We here illustrate innovative strategies using five challenging targets and variants thereof. Two libraries of 90 and 1056 fragments were screened using two different flow-based SPR biosensor systems, allowing different experimental approaches. Practical considerations and procedures accounting for the characteristics of the proteins and libraries, and that increase robustness, sensitivity, throughput and versatility are highlighted.

Portable-Surface Plasmon Resonance Biosensor Immunoassays for the Human Serum Albumin Detection

2013 ◽  
Vol 562-565 ◽  
pp. 408-411 ◽  
Author(s):  
Hui Li ◽  
Lu Lu Zhang ◽  
Hao Yuan Cai ◽  
Xing Chen ◽  
Jian Hai Sun ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Surface Plasmon Resonance ◽  
Human Serum Albumin ◽  
Light Intensity ◽  
Serum Albumin ◽  
Human Serum ◽  
Label Free ◽  
Detection Range ◽  
Spr Biosensor ◽  
Detection Assay

The screening of urine human serum albumin (HSA) is significant for the early diagnosis and intervention of the nephropathia. This study developed sensitive HSA detection assay with a self-made portable-SPR biosensor based on the reflection light intensity interrogation. Anti-HSA monoclonal antibody was immobilized onto the gold sensing area and a directive assay for HSA was developed. A detection range of 1–100μg/mL HSA was found. The described label-free, real-time immunoassay for the quantification of HSA in urine is a rapid, sensitive, alternative method available for the clinic detection.

High Sensitivity of Phase-based Surface Plasmon Resonance in Nano-cylinder Array

PIERS Online ◽  
2008 ◽  
Vol 4 (7) ◽  
pp. 746-750 ◽  
Author(s):  
Bing-Hung Chen ◽  
Yih-Chau Wang ◽  
Jia-Hng Lin
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
High Sensitivity ◽  
Cylinder Array

scholarly journals Comprehensive Metabolomic Analysis Reveals Dynamic Metabolic Reprogramming in Hep3B Cells with Aflatoxin B1 Exposure

Toxins ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 384
Author(s):  
Shufeng Wang ◽  
Xin Yang ◽  
Feng Liu ◽  
Xinzheng Wang ◽  
Xuemin Zhang ◽  
...  
Keyword(s):  
Aflatoxin B1 ◽  
Drug Targets ◽  
High Sensitivity ◽  
Tca Cycle ◽  
Acid Synthesis ◽  
Metabolic Reprogramming ◽  
Pyrimidine Metabolism ◽  
Rapid Separation ◽  
Hexosamine Pathway ◽  
Hep3b Cells

Hepatitis B virus (HBV) infection and aflatoxin B1 (AFB1) exposure have been recognized as independent risk factors for the occurrence and development of hepatocellular carcinoma (HCC), but their combined impacts and the potential metabolic mechanisms remain poorly characterized. Here, a comprehensive non-targeted metabolomic study was performed following AFB1 exposed to Hep3B cells at two different doses: 16 μM and 32 μM. The metabolites were identified and quantified by an ultra-performance liquid chromatography-mass spectrometry (UPLC-MS)-based strategy. A total of 2679 metabolites were identified, and 392 differential metabolites were quantified among three groups. Pathway analysis indicated that dynamic metabolic reprogramming was induced by AFB1 and various pathways changed significantly, including purine and pyrimidine metabolism, hexosamine pathway and sialylation, fatty acid synthesis and oxidation, glycerophospholipid metabolism, tricarboxylic acid (TCA) cycle, glycolysis, and amino acid metabolism. To the best of our knowledge, the alteration of purine and pyrimidine metabolism and decrease of hexosamine pathways and sialylation with AFB1 exposure have not been reported. The results indicated that our metabolomic strategy is powerful to investigate the metabolome change of any stimulates due to its high sensitivity, high resolution, rapid separation, and good metabolome coverage. Besides, these findings provide an overview of the metabolic mechanisms of the AFB1 combined with HBV and new insight into the toxicological mechanism of AFB1. Thus, targeting these metabolic pathways may be an approach to prevent carcinogen-induced cancer, and these findings may provide potential drug targets for therapeutic intervention.

scholarly journals Reusable TiN Substrate for Surface Plasmon Resonance Heterodyne Phase Interrogation Sensor

Nanomaterials ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 1325 ◽  
Author(s):  
Ru-Jing Sun ◽  
Hung Ji Huang ◽  
Chien-Nan Hsiao ◽  
Yu-Wei Lin ◽  
Bo-Huei Liao ◽  
...  
Keyword(s):  
Refractive Index ◽  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
High Sensitivity ◽  
Nanorod Array ◽  
Glass Substrates ◽  
Sensing Applications ◽  
Bulk Charge ◽  
Phase Interrogation

A TiN-based substrate with high reusability presented high-sensitivity refractive index measurements in a home-built surface plasmon resonance (SPR) heterodyne phase interrogation system. TiN layers with and without additional inclined-deposited TiN (i-TiN) layers on glass substrates reached high bulk charge carrier densities of 1.28 × 1022 and 1.91 × 1022 cm−3, respectively. The additional 1.4 nm i-TiN layer of the nanorod array presented a detection limit of 6.1 × 10−7 RIU and was higher than that of the 46 nm TiN layer at 1.2 × 10−6 RIU when measuring the refractive index of a glucose solution. Furthermore, the long-term durability of the TiN-based substrate demonstrated by multiple processing experiments presented a high potential for various practical sensing applications.

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