Characterization of botulinum neurotoxin type A subtypes by immunocapture enrichment and liquid chromatography–tandem mass spectrometry

Morineaux, V.; Mazuet, Christelle; Hilaire, Didier; Enche, Julien; Popoff, Michel R.

doi:10.1007/s00216-015-8707-1

Cited by 20 publications

(20 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Neutralization tests were performed using crude acid-precipitated toxins from strain Hall (type A1) and strain BL6 (type B2) [23,54]. Toxin preparations were diluted in 50 mM phosphate buffer pH 6.3 containing 0.2% gelatin (PB-G) and calibrated to approximately 10x mouse 50% lethal doses (10 MLD 50 ) per mL.…”

Section: Methodsmentioning

confidence: 99%

Development of Germline-Humanized Antibodies Neutralizing Botulinum Neurotoxin A and B

et al. 2016

Self Cite

View full text Add to dashboard Cite

Botulinum neurotoxins (BoNTs) are counted among the most toxic substances known and are responsible for human botulism, a life-threatening disease characterized by flaccid muscle paralysis that occurs naturally by food poisoning or colonization of the gastrointestinal tract by BoNT-producing clostridia. To date, 7 serologically distinct serotypes of BoNT (serotype A-G) are known. Due to the high toxicity of BoNTs the Centers for Disease Control and Prevention (CDC) have classified BoNTs as category A agent, including the six biological agents with the highest potential risk of use as bioweapons. Well tolerated antibodies neutralizing BoNTs are required to deal with the potential risk. In a previous work, we described the development of scFv and scFv-Fc (Yumab) from macaque origin (Macaca fascicularis) neutralizing BoNT/A and B by targeting the heavy and light chain of each serotype. In the present study, we humanized the macaque antibodies SEM120-IIIC1 (anti-BoNT/A light chain), A1HC38 (anti-BoNT/A heavy chain), BLC3 (anti-BoNT/B light chain) and B2-7 (anti-BoNT/B heavy chain) by germline-humanization to obtain a better potential immunotolerance in humans. We increased the Germinality Index (GI) of SEM120-IIIC1 to 94.5%, for A1HC38, to 95% for BLC3 and to 94.4% for B2-7. Furthermore, the neutralization efficacies of the germline-humanized antibodies were analyzed in lethal and non-lethal in vivo mouse assays as full IgG. The germline-humanized IgGs hu8SEM120-IIIC1, hu8A1HC38, hu8BLC3 and hu8B2-7 were protective in vivo, when anti-heavy and anti-light chain antibodies were combined. The synergistic effect and high humanness of the selected IgGs makes them promising lead candidates for further clinical development.

show abstract

Section: Methodsmentioning

confidence: 99%

Development of Germline-Humanized Antibodies Neutralizing Botulinum Neurotoxin A and B

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Morineaux et al [40] recently described a MS method that employs immunocapture enrichment by antibodies specific for BoNT/A-L chains. The enriched analyte is then analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a triple quadrupole mass spectrometer (QqQ) in multiple reaction monitoring (MRM) mode.…”

Section: Mass Spectrometry-based Methods To Detect Toxinsmentioning

confidence: 99%

Technologies for Detecting Botulinum Neurotoxins in Biological and Environmental Matrices

Cheng¹,

Land²,

Tam³

et al. 2016

Significance, Prevention and Control of Food Related Diseases

View full text Add to dashboard Cite

Abstract"iomonitoring of food and environmental matrices is critical for the rapid and sensitive diagnosis, treatment, and prevention of diseases caused by toxins. The U.S. Centers for Disease Control and Prevention CDC has noted that toxins from bacteria, fungi, algae, and plants present an ongoing public health threat, especially since some of these toxins could compromise security of the food supply. "otulinum neurotoxins "oNTs , produced by Clostridium spp., are among those bacterial toxins that pose life-threatening danger to humans. "oNTs inhibit the release of acetylcholine at peripheral cholinergic nerve terminals and cause flaccid paralysis. "oNTs are grouped in seven serotypes and many subtypes within these groups. Rapid and accurate identification of these toxins in contaminated food as well as in environmental matrices can help direct treatment. Herein, we discuss current methods to detect "oNTs with a focus on how these technologies have been used to identify toxins in various food and environmental matrices. We also discuss the emergence of new serotypes and subtypes of "oNTs and the increasing number of cases of botulism in wildlife. Finally, we consider how environmental changes impact food safety for humans and present new challenges for detection technology.

show abstract

“…MS‐based methods have also been effectively applied to the identification and specific detection of biological agents by analysis of intact proteins and/or tryptic digests from bacterial cells . The recent literature also indicates that the quest for specific and reliable markers of pathogenic species, such as B. anthracis or Y. pestis, needs to be pursued especially for their unambiguous discrimination from closely related nonpathogenic species.…”

Section: Detection Of Microorganisms and Toxins In Environmental And mentioning

confidence: 99%

“…[26] MS-based methods have also been effectively applied to the identification and specific detection of biological agents by analysis of intact proteins and/or tryptic digests from bacterial cells. [30][31][32][33][34][35] The recent literature also indicates that the quest for specific and reliable markers of pathogenic species, such as B. anthracis or Y. pestis, needs to be pursued especially for their unambiguous discrimination from closely related nonpathogenic species. As an example, using a high-resolution/high mass accuracy LTQ-Orbitrap instrument, a combined bottom-up and top-down proteomics methodology has been set up to identify specific markers of B. anthracis spores.…”

Section: Detection Of Microorganisms and Toxins In Environmental And mentioning

confidence: 99%

Mass spectrometry for the detection of bioterrorism agents: from environmental to clinical applications

Duriez¹,

Armengaud

Fenaille

et al. 2016

J. Mass Spectrom.

View full text Add to dashboard Cite

In the current context of international conflicts and localized terrorist actions, there is unfortunately a permanent threat of attacks with unconventional warfare agents. Among these, biological agents such as toxins, microorganisms, and viruses deserve particular attention owing to their ease of production and dissemination. Mass spectrometry (MS)-based techniques for the detection and quantification of biological agents have a decisive role to play for countermeasures in a scenario of biological attacks. The application of MS to every field of both organic and macromolecular species has in recent years been revolutionized by the development of soft ionization techniques (MALDI and ESI), and by the continuous development of MS technologies (high resolution, accurate mass HR/AM instruments, novel analyzers, hybrid configurations). New possibilities have emerged for exquisite specific and sensitive detection of biological warfare agents. MS-based strategies for clinical application can now address a wide range of analytical questions mainly including issues related to the complexity of biological samples and their available volume. Multiplexed toxin detection, discovery of new markers through omics approaches, and identification of untargeted microbiological or of novel molecular targets are examples of applications. In this paper, we will present these technological advances along with the novel perspectives offered by omics approaches to clinical detection and follow-up.

show abstract

Characterization of botulinum neurotoxin type A subtypes by immunocapture enrichment and liquid chromatography–tandem mass spectrometry

Cited by 20 publications

References 31 publications

Development of Germline-Humanized Antibodies Neutralizing Botulinum Neurotoxin A and B

Development of Germline-Humanized Antibodies Neutralizing Botulinum Neurotoxin A and B

Technologies for Detecting Botulinum Neurotoxins in Biological and Environmental Matrices

Mass spectrometry for the detection of bioterrorism agents: from environmental to clinical applications

Contact Info

Product

Resources

About