Novel methods for biocatalyst screening

Wahler, Denis; Reymond, Jean‐Louis

doi:10.1016/s1367-5931(00)00184-8

Cited by 155 publications

(54 citation statements)

References 49 publications

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“…The absorbance is dependent on both [G] t and ee because the right chamber is occupied by the chiral host (S,S)-2 [Eq. (2)]. Putting these two equations together, the values of [G] t and ee can be solved.…”

Section: Resultsmentioning

confidence: 99%

“…Such a signal is related to [G] t and ee by way of a mathematical function of solution equilibrium constants and molar absorptivities ([Equation (2)], Scheme 1). [11] Because [G] t is known from the measurement using the achiral host H, the ee of the sample can be obtained by solving Equation (2). We have used an arylboronic acid based eIDA to quantitatively analyze chiral a-hydroxycarboxylates and diols.…”

Section: Introductionmentioning

confidence: 99%

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Two Methods for the Determination of Enantiomeric Excess and Concentration of a Chiral Sample with a Single Spectroscopic Measurement

Zhu

Shabbir

Anslyn

2006

Chemistry A European J

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The previously established enantioselective indicator–displacement assays (eIDAs) for the determination of concentration and enantiomeric excess (ee) require two spectroscopic measurements for each chiral sample. To further simplify the operation of eIDAs, we now introduce two innovative analytical methods, both of which utilize a dual‐chamber quartz cuvette, which reduces the number of spectroscopic measurements from two to one. An attractive feature of this cuvette is that the concentration‐ and ee‐dependent absorption data can be collected at the isosbestic points or transparent regions of the spectra recorded in each individual chamber, thereby reflecting optical changes that occur in the other chamber. Therefore, two independent equations, which are needed to solve the values of the two independent variables—concentration and ee—can be established with only a single spectroscopic measurement. The first method takes advantage of this feature in conjunction with a judicious choice of indicator/host combinations to generate concentration‐ and ee‐dependent calibration curves. Our second method removes the requirement to measure equilibrium constants and molar absorptivities altogether through the use of artificial neural networks (ANNs). The most frequently used three‐layer feed‐forward network is generated, which relates the absorption data to concentration and ee of the samples by training with a back propagation procedure. Here, the data collection is not limited to the isosbestic points or transparent regions. Both approaches enabled accurate and rapid determination of concentration and ee of chiral samples. The technology removes the relative difficulty, which is the need for two separate measurements for concentration and ee respectively, of analyzing chiral samples compared to achiral samples. When implemented in a high‐throughput format, this technology should greatly facilitate the discovery of asymmetric catalysts in the same way as conventional high‐throughput screening assays.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Two Methods for the Determination of Enantiomeric Excess and Concentration of a Chiral Sample with a Single Spectroscopic Measurement

Zhu

Shabbir

Anslyn

2006

Chemistry A European J

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“…This may also occur when abzymes are obtained as idiotypic or anti-idiotypic antibodies against the enzyme, as observed for AChE-like antibodies. [33][34][35][36] These were the reasons why we changed the experimental approach to the isolation of an AchE-like scFv, integrating the screening by genetic selection 37,38 with the bait-and-switch strategy 39,40 for the elicitation of antibody catalysts. Phage libraries represent an excellent tool for antibody selection as demonstrated by the drugs already produced on the market.…”

Section: Discussionmentioning

confidence: 99%

Selection of a human butyrylcholinesterase-like antibody single-chain variable fragment resistant to AChE inhibitors from a phage library expressed inE. coli

Podestà¹,

Rossi²,

Massarelli³

et al. 2014

mAbs

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“…In general, these techniques have a limited dynamic (concentration) range. In addition, UV-Vis and fluorescence spectroscopies require an optical activity of the reactants or products and, in their absence, the analysis is more complicated as chromogenic and fluorogenic substrates are needed [8][9][10]. Therefore, discontinuous methods are widely used for the monitoring of chemical reactions, including chromatography, mass spectrometry (MS) and capillary electrophoresis [9].…”

Section: Introductionmentioning

confidence: 99%

Ultrasonic Monitoring of Biocatalysis in Solutions and Complex Dispersions

Buckin

Altas

2017

Catalysts

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Abstract:The rapidly growing field of chemical catalysis is dependent on analytical methods for non-destructive real-time monitoring of chemical reactions in complex systems such as emulsions, suspensions and gels, where most analytical techniques are limited in their applicability, especially if the media is opaque, or if the reactants/products do not possess optical activity. High-resolution ultrasonic spectroscopy is one of the novel technologies based on measurements of parameters of ultrasonic waves propagating through analyzed samples, which can be utilized for real-time non-invasive monitoring of chemical reactions. It does not require optical transparency, optical markers and is applicable for monitoring of reactions in continuous media and in micro/nano bioreactors (e.g., nanodroplets of microemulsions). The technology enables measurements of concentrations of substrates and products over the whole course of reaction, analysis of time profiles of the degree of polymerization and molar mass of polymers and oligomers, evolutions of reaction rates, evaluation of kinetic mechanisms, measurements of kinetic and equilibrium constants and reaction Gibbs energy. It also provides tools for assessments of various aspects of performance of catalysts/enzymes including inhibition effects, reversible and irreversible thermal deactivation. In addition, ultrasonic scattering effects in dispersions allow real-time monitoring of structural changes in the medium accompanying chemical reactions.

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Novel methods for biocatalyst screening

Cited by 155 publications

References 49 publications

Two Methods for the Determination of Enantiomeric Excess and Concentration of a Chiral Sample with a Single Spectroscopic Measurement

Two Methods for the Determination of Enantiomeric Excess and Concentration of a Chiral Sample with a Single Spectroscopic Measurement

Selection of a human butyrylcholinesterase-like antibody single-chain variable fragment resistant to AChE inhibitors from a phage library expressed inE. coli

Ultrasonic Monitoring of Biocatalysis in Solutions and Complex Dispersions

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