A Protein Biosensor for Lactate

D’Auria, Sabato; Gryczyński, Zygmunt; Gryczyński, Ignacy; Rossi, Mosè; Lakowicz, Joseph R.

doi:10.1006/abio.2000.4611

Cited by 29 publications

(16 citation statements)

References 30 publications

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“…In particular, a number of biosensors based on enzymes such as lactate dehydrogenase or lactate oxidase have been developed. The output signals from these biosensors vary, but include spectroscopic (Shah et al, 2007), fluorescent (D'Auria et al, 2000; Li et al, 2009; Trettnak and Wolfbeis, 1989), chemiluminescent (Martinez‐Olmos et al, 2009; Roda et al, 2014; Zhou et al, 2014), or amperometric detection (Haccoun et al, 2006; Hasebe et al, 2005; Male et al, 1997; Parra et al, 2006; Weber et al, 2006), with the latter as the dominant detection modality (recently reviewed in (Rathee et al, 2016)). Novel methods of immobilization to improve detection have also been explored (Haccoun et al, 2006; Hasebe et al, 2005; Pagan et al, 2015; Tu et al, 2016; Xu et al, 2005; Yang et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Whole‐cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Goers

Ainsworth

Goey

et al. 2017

Biotech & Bioengineering

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Many high‐value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimize the productivity of these cultures it is important to monitor cellular metabolism, for example the utilization of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole‐cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity, and robustness. Importantly, our whole‐cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole‐cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalize on the wealth of genetic operons for metabolite sensing available in nature for the development of other whole‐cell biosensors. Biotechnol. Bioeng. 2017;114: 1290–1300. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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Section: Introductionmentioning

confidence: 99%

Whole‐cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Goers

Ainsworth

Goey

et al. 2017

Biotech & Bioengineering

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“…l-Lactate is a key metabolite, the detection of which is important across a number of disciplines, including clinical diagnostics [10,11] and sports medicine. [12,13] The integration of boronic acids into polymers has been reported previously [14] and more recently a hydrogel was generated that enabled the holographic determination of l-lactate.…”

Section: Introductionmentioning

confidence: 99%

Complexation of L‐Lactate with Boronic Acids: A Solution and Holographic Analysis

Sartain

Yang

Lowe

2008

Chemistry A European J

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Boronic acids have been used as receptors for the detection of diols and alpha-hydroxy acids. The incorporation of 3-acrylamide phenyl boronic acid (3-APB) into a hydrogel generates a suitably responsive and fully reversible holographic sensor for L-lactate. However, it was also found that the use of 3-APB resulted in the sensor being responsive towards a number of other compounds containing two hydroxy groups. This report details the further investigation into the reaction between L-lactate and three boronic acid-based receptors, both in the holograms and in solution, in order to establish the mechanism of binding. A novel boronate receptor is proposed based on this understanding.

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“…17 Nevertheless, there was some leaching of the enzyme and cofactor from the matrix. Lactate dehydrogenase has also been employed in a sensor described by D'Auria et al, 18 but here the dye 8-anilino-1-naphthalene was noncovalently bound to the enzyme as a reporter, displaying a 40% decrease in fluorescence emission on binding to lactate.…”

Section: Monitoring Tissue Oxygenation: Lactic Acidmentioning

confidence: 99%

Biosensors for Monitoring Metabolites in Clinical Medicine

Pickup

2007

Handbook of Biosensors and Biochips

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Biosensors for small‐molecular‐weight metabolites are finding increasing use in medicine for rapid, reagentless point‐of‐care measurement and continuous in vivo monitoring. Though glucose is the most well‐established clinical biosensor application, others include urea and creatinine for assessing renal function; lactic acid for monitoring tissue oxygenation; uric acid for assessing nucleic acid metabolism; cholesterol, triglyceride, and ketones for monitoring lipid metabolism; and bilirubin for liver and blood disorders. Many metabolite biosensors are enzyme electrodes, based on immobilized oxidases, hydrolases, or other enzymes, with amperometric or potentiometric electrochemical reaction monitoring, but other bioreceptors such as computer‐designed proteins and other reaction detection methods such as fluorescence are in use.

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A Protein Biosensor for Lactate

Cited by 29 publications

References 30 publications

Whole‐cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Whole‐cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Complexation of L‐Lactate with Boronic Acids: A Solution and Holographic Analysis

Biosensors for Monitoring Metabolites in Clinical Medicine

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