News Release

Cleanascite™ Employed to Investigate Membrane Vesicle Influences on Bacterial Biofilms

MONMOUTH JUNCTION, NJ, January 27, 2023 -- Biotech Support Group reports on an article, describing the simplicity and efficiency of their lipid binding technology to determine the influence of probiotic membrane vesicles on antimicrobial activity.

da Silva Barreira, David, et al. "Membrane vesicles released by Lacticaseibacillus casei BL23 inhibit the biofilm formation of Salmonella Enteritidis." Scientific Reports 13.1 (2023): 1163.

Biofilms are defined as spatially organized communities of microorganisms embedded in a self-produced matrix of extracel- lular polymeric substances (EPS), and represent a major concern in the food industry and healthcare. They contain a diversity of elements including polysaccharides, proteins, nucleic acids, lipids and membrane vesicles (MVs) (also referred to as extracellular vesicles), contributing to cellular heterogeneity and resistance to antibiotic treatments. However, the antimicrobial activity of probiotic MVs remains to be studied. The “Generally Regarded as Safe” (GRAS) status of L. casei probiotics make them good candidates to investigate antimicrobial activities against pathogens such as Salmonella enterica.

The article states “To investigate the effect of L. casei BL23 on S. Enteritidis biofilm formation, we first quantified the biofilm biomass formed after treatment with several fractions of L. casei BL23 cell-free supernatant (LC-CFS). To this end, the LC-CFS and the growth medium of the bacteria (i.e. MRS) were fractionated by size-exclusion ultrafiltration to obtain several fractions ranging from 3 kDa to over 100 kDa”. 

“Next, we decided to remove all lipids from LC-CFS > 100 and LC-CFS > 3 fractions and their corresponding controls (MRS > 100 and MRS > 3) using a lipid removal reagent…Cleanascite; following the manufacturer’s instructions.” “The antibiofilm activity of the delipidated fractions (Lipid −) was then compared to the initial fractions (Lipid +) by crystal violet staining (Fig.)…we saw a significant increase in biomass formation with the delipidated LC-CFS > 3 fraction compared to the control (MRS > 3) and the untreated fractions.” The report showed that the cell-free supernatant (LC-CFS) and the cell lysate of L. casei BL23 have a strong antibiofilm activity against S. Enteritidis, whereas live cells have no significant effect. Furthermore, it was demonstrated that membrane vesicles (LC-MVs) released by L. casei BL23 contribute to the antibiofilm activity of LC-CFS but have no impact on bacterial growth. In addition, it was showed that LC-MVs have a strong antibiofilm effect at the early stage of biofilm formation and no effect on established biofilms of S. Enteritidis. Finally, it was demonstrated that proteins associated with LC-MVs are responsible for the antibiofilm effects of the vesicles, and two peptidoglycan hydrolases (PGHs) were found to be involved in vesicle activities.

“This is now the second published report showing that Cleanascite™ efficiently removes membrane and extracellular vesicles, to determine their influence on cellular activity. Unlike alternative lipid removal methods that use solvents, Cleanascite™ is an aqueous suspension product and so it is very compatible with cellular response assays such as those used here. This ultimately helps with investigations such as this, that need to determine whether or not lipids, or factors associated with lipid such as extracellular vesicles, impart phenotypic changes to cells.” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group. 

Link to view our case study, Diverse technologies available for researchers to selectively bind or enrich exosomes and extracellular vesicles.

Link to view application report entitled “LIPID REMOVAL & CELL RESPONSE APPLICATIONS”

For more information visit: Cleanascite™ Lipid Removal Reagent and Clarification, at
http://www.biotechsupportgroup.com/Cleanascite-Lipid-Removal-Reagent-p/x2555.htm

Keywords: Membrane vesicles, extracellular vesicles, exosomes, Biofilms, Cleanascite™, Probiotics, extracellular vesicle removal

About Biotech Support Group LLC

Converging with cultural and technological disruptions forthcoming in healthcare, Biotech Support Group develops methods for cost effective and efficient sample prep essential for these expanding markets. Following a tiered business strategy, the company continues its growth in the consumable research products area supporting the rapidly expanding installation of LC-MS instrument and computational infrastructure. For this market, key products include: AlbuVoid™ and AlbuSorb™ for albumin depletion, Cleanascite™ for lipid adsorption, HemogloBind™ and HemoVoid™ for hemoglobin removal. From these innovations, the company has acquired knowledgebase and biomarker intellectual property assets that support discoveries of protein markers from blood, with special emphasis on early detection and personalized medical decisions for cancer patients. For more information, go to http://www.biotechsupportgroup.com.

For Business Development, contact: Matthew Kuruc 732-274-2866, mkuruc@biotechsupportgroup.com