Screening and selection of synthetic peptides for a novel and optimized endotoxin detection method

Mujika, Maite; Zuzuarregui, Ana; Sánchez-Gómez, Susana; Tejada, Guillermo Martínez de; Arana, Sergio; Pérez-Lorenzo, E.

doi:10.1016/j.jbiotec.2014.06.018

Cited by 11 publications

(12 citation statements)

References 37 publications

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“…The pathology of the discomforts ( i.e. , severe sepsis and septic shock) is the repercussion of an unbalanced response of the host to pathogen-associated molecular patterns (PAMPs) [42]. These PAMPs may be of immunological significance if the endotoxin concentration in the mammalian cell is relatively low.…”

Section: Pathophysiological Consequencesmentioning

confidence: 99%

“…Generally, the injection of LPS into surrounding medium takes place during cell death, cell growth and cell division [3,18], shedding, proliferation or degradation [61], and even in the use of antibiotics [2,42]. For instance, the lysis of bacterial membranes when harvesting recombinant proteins from E. coli leads to the disengagement of endotoxin from the membranes and results in the contamination of the product [62].…”

Section: Lps and Biomolecule Interactionmentioning

confidence: 99%

“…The LAL gel test is believed to be sensitive to the presence of endotoxin; however, it significantly lacks robustness and repeatability, especially in the detection of material-bound LPS and other microbial contaminations [64]. It is strongly sensitive to disturbances in the form of pH, proteases, divalent cations, chelators, anticoagulants and serum [42]. Moreover, this assay is known to be laborious and liable to batch-to-batch variability [42].…”

Section: Endotoxin Detection and Quantificationmentioning

confidence: 99%

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Elucidating endotoxin-biomolecule interactions with FRET: extending the frontiers of their supramolecular complexation

et al. 2017

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Endotoxin has been one of the topical chemical contaminants of major concern to researchers, especially in the field of bioprocessing. This major concern of researchers stems from the fact that the presence of Gram-negative bacterial endotoxin in intracellular products is unavoidable and requires complex downstream purification steps. For instance, endotoxin interacts with recombinant proteins, peptides, antibodies and aptamers and these interactions have formed the foundation for most biosensors for endotoxin detection. It has become imperative for researchers to engineer reliable means/techniques to detect, separate and remove endotoxin, without compromising the quality and quantity of the end-product. However, the underlying mechanism involved during endotoxin-biomolecule interaction is still a gray area. The use of quantitative molecular microscopy that provides high resolution of biomolecules is highly promising, hence, may lead to the development of improved endotoxin detection strategies in biomolecule preparation. Förster resonance energy transfer (FRET) spectroscopy is one of the emerging most powerful tools compatible with most super-resolution techniques for the analysis of molecular interactions. However, the scope of FRET has not been well-exploited in the analysis of endotoxin-biomolecule interaction. This article reviews endotoxin, its pathophysiological consequences and the interaction with biomolecules. Herein, we outline the common potential ways of using FRET to extend the current understanding of endotoxin-biomolecule interaction with the inference that a detailed understanding of the interaction is a prerequisite for the design of strategies for endotoxin identification and removal from protein milieus.

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Section: Pathophysiological Consequencesmentioning

confidence: 99%

Section: Lps and Biomolecule Interactionmentioning

confidence: 99%

Section: Endotoxin Detection and Quantificationmentioning

confidence: 99%

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Elucidating endotoxin-biomolecule interactions with FRET: extending the frontiers of their supramolecular complexation

et al. 2017

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“…cannot be assayed by BET. Despite scientist having proposed several in vitro methods which are faster and more easily automated than the original LAL-tests of Cooper and Mills, the latter still remains the only official in the main pharmacopeias [ 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

Endotoxins from a Pharmacopoeial Point of View

Franco

García-Recio

Jiménez

et al. 2018

Toxins

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A pyrogen is a substance that causes fever after intravenous administration or inhalation. Gram negative endotoxins are the most important pyrogens to pharmaceutical laboratories. In the International, United States, Japanese and European Pharmacopoeias, there are two official methods to evaluate pyrogenicity—that is, the bacterial endotoxin test, and the pyrogen test. The main objective of this review is to compare the monographs of each test among the different Pharmacopeias, to detect similarities and differences. The former can be considered fully harmonized, and only non-significant differences were detected. The latter, which is the only available assay for some products and formulations to demonstrate apyrogenicity, shows large differences, which should be considered.

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“…These methods depend on selection of a recognition element, especially a bio-receptor or ligand. Several systems have been reported for the detection of LPS that use anti-LPS antibodies8 or antibiotics such as Polymyxin B91011, and molecules designed specifically for LPS neutralization, namely aptamers121314, LPS binding proteins1516171819 or peptides2021, and other synthetic LPS recognizing molecules22232425 as the recognition element. In addition to using natural and synthetic molecules with LPS affinity, living cells to detect LPS have been designed726.…”

mentioning

confidence: 99%

High-throughput living cell-based optical biosensor for detection of bacterial lipopolysaccharide (LPS) using a red fluorescent protein reporter system

Jiang

Shao³

et al. 2016

Sci Rep

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Due to the high toxicity of bacterial lipopolysaccharide (LPS), resulting in sepsis and septic shock, two major causes of death worldwide, significant effort is directed toward the development of specific trace-level LPS detection systems. Here, we report sensitive, user-friendly, high-throughput LPS detection in a 96-well microplate using a transcriptional biosensor system, based on 293/hTLR4A-MD2-CD14 cells that are transformed by a red fluorescent protein (mCherry) gene under the transcriptional control of an NF-κB response element. The recognition of LPS activates the biosensor cell, TLR4, and the co-receptor-induced NF-κB signaling pathway, which results in the expression of mCherry fluorescent protein. The novel cell-based biosensor detects LPS with specificity at low concentration. The cell-based biosensor was evaluated by testing LPS isolated from 14 bacteria. Of the tested bacteria, 13 isolated Enterobacteraceous LPSs with hexa-acylated structures were found to increase red fluorescence and one penta-acylated LPS from Pseudomonadaceae appeared less potent. The proposed biosensor has potential for use in the LPS detection in foodstuff and biological products, as well as bacteria identification, assisting the control of foodborne diseases.

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Screening and selection of synthetic peptides for a novel and optimized endotoxin detection method

Cited by 11 publications

References 37 publications

Elucidating endotoxin-biomolecule interactions with FRET: extending the frontiers of their supramolecular complexation

Elucidating endotoxin-biomolecule interactions with FRET: extending the frontiers of their supramolecular complexation

Endotoxins from a Pharmacopoeial Point of View

High-throughput living cell-based optical biosensor for detection of bacterial lipopolysaccharide (LPS) using a red fluorescent protein reporter system

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