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1.
J Proteome Res ; 13(3): 1167-76, 2014 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-24450425

RESUMO

We have developed a simple method for the release and isolation of glycoprotein N-glycans from whole-cell lysates using less than a million cells, for subsequent implementation with mass spectrometric analysis. Cellular protein extracts prepared using SDS solubilization were sequentially treated in a membrane filter device to ultimately release glycans enzymatically using PNGase F in the volatile buffer ammonium bicarbonate. The released glycans are recovered in the filtrate following centrifugation and typically permethylated prior to mass spectrometric analysis. We call our method "filter-aided N-glycan separation" and have successfully applied it to investigate N-glycan profiles of wild-type and mutant Chinese hamster ovary cells. This method is readily multiplexed and, because of the small numbers of cells needed, is compatible with the analysis of replicate samples to assess the true nature of glycan variability in tissue culture samples.


Assuntos
Extratos Celulares/química , Fracionamento Químico/métodos , Glicoproteínas/química , Polissacarídeos/isolamento & purificação , Animais , Células CHO , Sequência de Carboidratos , Cricetulus , Filtração , Dados de Sequência Molecular , Peptídeo-N4-(N-acetil-beta-glucosaminil) Asparagina Amidase/química , Polissacarídeos/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
2.
Int J Anal Chem ; 2024: 8581029, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38312506

RESUMO

It has been well-established that mycotoxins are poisonous chemical metabolites secreted by certain molds. Some of them significantly affect the health of humans and livestock. Increasing attention is now being paid to uncovering and identifying mycotoxins' presence in the building's environment. However, the main challenge remains in suitable and reliable analytical methods for their identification and detection in infected structures. GC-MS and LC-MS/MS techniques have been used extensively for mycotoxin analysis, and advancement in these techniques enabled a more comprehensive range of mycotoxins to be detected. As such, this study aimed to address a brief overview of various phenomena of existing sample collection, preparation, and analysis to detect mycotoxins in the building's environment. This scoping review includes articles from 2010 to 2020 available from PubMed, Scopus, Cochrane, Wiley, Google Scholar, and ScienceDirect. Duplicate articles were removed, and exclusion criteria were applied to eliminate unrelated studies, resulting in 14 eligible articles. The present study provides an overview of mycotoxin analysis by GC-MS and LC-MS/MS in buildings. Many techniques are available for analyzing and detecting multiple mycotoxins using these methods. Future efforts would focus on rapid assays and tools enabling measuring a broader range of mycotoxins in a single matrix and lower detection limits. In addition, it would assist future findings on new techniques and mycotoxins that existed in the building's environment.

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