Rational design of a turn-on fluorescent sensor for α-ketoglutaric acid in a microfluidic chip

2014 ◽  
Vol 5 (10) ◽  
pp. 4012-4016 ◽  
Author(s):  
Pengwei Jin ◽  
Changhong Jiao ◽  
Zhiqian Guo ◽  
Ye He ◽  
Shiqin Zhu ◽  
...  
Keyword(s):  
High Throughput ◽  
Microfluidic Chip ◽  
High Throughput Screening ◽  
Rational Design ◽  
Fluorescent Sensor ◽  
Fluorescent Chemosensors ◽  
Turn On

A rational design of turn-on fluorescent chemosensors for monitoring α-ketoglutaric acid has been developed with a microfluidic chip, indicative of a potential platform for high-throughput screening and monitoring of kinetics, especially in biological fields.

scholarly journals High-throughput screening and rational design of biofunctionalized surfaces with optimized biocompatibility and antimicrobial activity

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Zhou Fang ◽  
Junjian Chen ◽  
Ye Zhu ◽  
Guansong Hu ◽  
Haoqian Xin ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
High Throughput ◽  
High Throughput Screening ◽  
Rational Design ◽  
Click Reaction ◽  
Reaction Times ◽  
Great Promise ◽  
Biomaterial Surfaces

AbstractPeptides are widely used for surface modification to develop improved implants, such as cell adhesion RGD peptide and antimicrobial peptide (AMP). However, it is a daunting challenge to identify an optimized condition with the two peptides showing their intended activities and the parameters for reaching such a condition. Herein, we develop a high-throughput strategy, preparing titanium (Ti) surfaces with a gradient in peptide density by click reaction as a platform, to screen the positions with desired functions. Such positions are corresponding to optimized molecular parameters (peptide densities/ratios) and associated preparation parameters (reaction times/reactant concentrations). These parameters are then extracted to prepare nongradient mono- and dual-peptide functionalized Ti surfaces with desired biocompatibility or/and antimicrobial activity in vitro and in vivo. We also demonstrate this strategy could be extended to other materials. Here, we show that the high-throughput versatile strategy holds great promise for rational design and preparation of functional biomaterial surfaces.

scholarly journals In Vivo, High-Throughput Selection of Thermostable Cyclohexanone Monooxygenase (CHMO)

Catalysts ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 935
Author(s):  
Sarah Maxel ◽  
Linyue Zhang ◽  
Edward King ◽  
Ana Paula Acosta ◽  
Ray Luo ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Rational Design ◽  
Random Mutagenesis ◽  
Residual Activity ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Coli Strain ◽  
Screening Tools ◽  
Cyclohexanone Monooxygenase

Cyclohexanone monooxygenase (CHMO) from Acinetobacter sp. NCIMB 9871 is characterized as having wide substrate versatility for the biooxidation of (cyclic) ketones into esters and lactones with high stereospecificity. Despite industrial potential, CHMO usage is restricted by poor thermostability. Limited high-throughput screening tools and challenges in rationally engineering thermostability have impeded CHMO engineering efforts. We demonstrate the application of an aerobic, high-throughput growth selection platform in Escherichia coli (strain MX203) for the discovery of thermostability enhancing mutations for CHMO. The selection employs growth for the easy readout of CHMO activity in vivo, by requiring nicotinamide adenine dinucleotide phosphate (NADPH)-consuming enzymes to restore cellular redox balance. In the presence of the native substrate cyclohexanone, variant CHMO GV (A245G-A288V) was discovered from a random mutagenesis library screened at 42 °C. This variant retained native activity, exhibited ~4.4-fold improvement in residual activity after 30 °C incubation, and demonstrated ~5-fold higher cyclohexanone conversion at 37 °C compared to the wild type. Molecular modeling indicates that CHMO GV experiences more favorable residue packing and supports additional backbone hydrogen bonding. Further rational design resulted in CHMO A245G-A288V-T415C with improved thermostability at 45 °C. Our platform for oxygenase evolution enabled the rapid engineering of protein stability critical for industrial scalability.

scholarly journals Challenges and Opportunities in High Throughput Screening: Implications for New Technologies

1998 ◽  
Vol 3 (1) ◽  
pp. 13-17 ◽  
Author(s):  
John Major
Keyword(s):  
Pharmaceutical Industry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Rational Design ◽  
Information Science ◽  
New Technologies ◽  
New Technology ◽  
Current Trend ◽  
Final Decision ◽  
Design Activities

In response to mounting competitive pressures, the current trend in the pharmaceutical industry is to shorten the time scale for all aspects of drug discovery. While advances in computation, structural chemistry, and molecular modeling are facilitating rational design activities, empirical screening continues to play a crucial role in lead identification. Because the ability to test large numbers of compounds quickly and efficiently can provide a competitive advantage, high throughput screening (HTS) has become a key tool in many companies. To achieve the necessary productivity, effective integration of compound supply, assay operation and data management is essential. HTS is a very high technology enterprise that must take full advantage of the latest advances in bioscience, biotechnology, engineering, and information science. There is a constant dilemma, however, in relation to when well-established, mature technologies should be replaced by new methods that promise to deliver spectacular advantages. The final decision must be based on weighing the promised benefit against the cost and risk. While huge challenges face the pharmaceutical industry, there are also opportunities for those companies that can identify and implement new technology effectively.

Rational design of a highly sensitive and selective “turn-on” fluorescent sensor for PO43− detection

2015 ◽  
Vol 44 (48) ◽  
pp. 20830-20833 ◽  
Author(s):  
Si-Quan Jiang ◽  
Zi-Yan Zhou ◽  
Shu-Ping Zhuo ◽  
Guo-Gang Shan ◽  
Ling-Bao Xing ◽  
...  
Keyword(s):  
Schiff Base ◽  
Rational Design ◽  
Fluorescent Sensor ◽  
Turn On ◽  
Base Unit ◽  
Highly Sensitive

An in situ-generated iron(iii) complex with a 1,8-naphthalene-based Schiff base unit has been rationally designed, which exhibits a highly selective response and excellent sensitivity for the turn-on detection of PO43− anions.

A Colorimetric and “Turn-On” Fluorescent Chemosensor for Metal Ion Based on Rhodamine Derivative

2014 ◽  
Vol 955-959 ◽  
pp. 267-270
Author(s):  
Hao Zhang ◽  
Xi Jiang
Keyword(s):  
Schiff Base ◽  
High Throughput ◽  
Chemical Sensors ◽  
High Throughput Screening ◽  
Metal Ion ◽  
Important Species ◽  
Rhodamine Derivative ◽  
Turn On ◽  
Highly Sensitive ◽  
Rhodamine Derivatives

The recognition and sensing of biologically and environmentally important species has emerged as a significant goal in the field of chemical sensors in recent years1. Fluorogenic methods in conjunction with suitable probes are preferable approaches for the measurement of these analytes because fluorimetry is rapidly performed, is nondestructive, is highly sensitive, is suitable for high-throughput screening applications2,3. We synthesized rhodamine derivatives compounds by a Schiff base reaction.

Sign in / Sign up

Export Citation Format

Share Document