A Review on Macroscale and Microscale Cell Lysis Methods

Islam, Mohammed Shehadul; Aryasomayajula, Aditya; Selvaganapathy, P. Ravi

doi:10.3390/mi8030083

Cited by 326 publications

(194 citation statements)

References 106 publications

(123 reference statements)

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“…Mechanical lysis of bacteria was selected as the method to lyse the cells, because it has the advantage to preserving the antigenic structure, as opposed to chemical methods like alkaline lysis which produce changes in protein structure [29]. All the lysates (individual or polyvalent) antigenic patterns were analyzed by SDS gel electrophoresis, protein and carbohydrate content and by a mass spectrometry.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Bacterial Lysates by Use of Matrix-Assisted Laser Desorptionâ€“ Ionization Time of Flight Mass Spectrometry Fingerprinting

Suárez¹,

Ferrara²,

Pirez³

et al. 2017

SOJB

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Bacterial lysates have for long been used to boost the immunological response to Respiratory Tract Infections (RTI) both in children and adults. They are prepared by growing bacteria usually associated with RTI, followed by chemical or mechanical disruption to prepare single bacterial lysates that are combined in the final product. Despite the wide range of applications, one drawback for their universal use is the difficulty to assure consistency in their composition given their particular form of preparation; thus there is a need for alternative analytical methods that ensures batch composition consistency. Here, we demonstrate that MALDI-TOF MS provides reliable and reproducible mass spectral fingerprints for bacterial lysates of S. pneumoniae, H. influenzae, K. pneumoniae, Staphylococcus spp. We also found that mechanical disruption provides markedly betterdefined fingerprints. Analysis of the formulated Polyvalent Bacterial Mechanical Lysate (PBML) also showed a characteristic spectrum. Overall, we found that mechanical lysis coupled to MS analysis allowed for accurate and highly sensible detection of key proteins in each bacterial lysate, a method that can be used to standardize batch-to-batch product composition. Applying this methodology to the production pipeline shall result in better products expanding the acceptance of these cost-effective tools to prevent respiratory tract pathologies.

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Section: Resultsmentioning

confidence: 99%

Characterization of Bacterial Lysates by Use of Matrix-Assisted Laser Desorptionâ€“ Ionization Time of Flight Mass Spectrometry Fingerprinting

Suárez¹,

Ferrara²,

Pirez³

et al. 2017

SOJB

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“…Cavitation's produce high energies around the cells leading to their breakdown and cell disruption. This leads to a high diffusion rates across the cell walls and better penetration [35].…”

Section: Scanning Electron Microscopymentioning

confidence: 99%

Comparative Evaluation of Soxhlet and Ultrasonics on the Structural Morphology and Extraction of Bioactive Compounds of Lemon (Citrus limon L.) Peel

Jagannath¹,

Biradar²

2019

J Food Chem Nanotechnol

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The relative recoveries of bioactive compounds and nutritional ingredients from lemon peel by ultrasound-assisted extraction (USAE/UAE) and traditional soxhlet extraction methods under different conditions of extraction were evaluated. The effect of these treatments on the morphological structure and tissue damage were assessed by scanning electron microscopy. The overall optimal conditions for maximum extraction of total phenolic content (TPC), total flavonoids content (TFC), vitamin C and anti-oxidant activity (AOA) using USAE (200 W output) was predicted using response surface methodology to be at the combined levels of 48 °C extraction temperature, 60 min extraction time and 80% duty cycle. Under these optimum conditions the corresponding predicted responses for TPC, TFC, vitamin C and AOA were 67.17 mg GAE/100 g, 4.52 mg CE/100 g, 26.46 mg/100 g and 61.68% respectively. USAE proved better as compared to soxhlet in the extraction of total phenolics, flavonoids, retention of vitamin C and showing antioxidant activity. In the case of USAE, the combination of factors for maximum extraction of total phenolics (88.06 mg GAE/100 g) was found to be at a temperature of 33 °C, 80% duty cycle and 40 min of treatment. With reference to maximum extraction of total flavonoids (13.92 mg CE/100 g) these conditions were 40 °C, 60% and 50 min and 40 °C, 100% duty cycle, 50 min of treatment for vitamin C (58.87 mg/100 g) contents respectively. The conditions which showed the maximum antioxidant activity (69.37%) was found to be 50 °C, 80% duty cycle and 40 min of extraction.

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“…Piercing the cell envelope would produce local cell damage and lysis of the bacterium. 26,30,31,60 The absence of the bacterial cell envelope in Figure 2B indicates that this damage was sufficient to stop the metabolic maintenance of the cell envelope or produce apoptosis. 60 In contrast to mathematical modeling in previous…”

Section: Resultsmentioning

confidence: 99%

<p>Correlative ex situ and Liquid-Cell TEM Observation of Bacterial Cell Membrane Damage Induced by Rough Surface Topology</p>

Banner

Firlar

Jakubonis

et al. 2020

IJN

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Background: Nanoscale surface roughness has been suggested to have antibacterial and antifouling properties. Several existing models have attempted to explain the antibacterial mechanism of nanoscale rough surfaces without direct observation. Here, conventional and liquid-cell TEM are implemented to observe nanoscale bacteria/surface roughness interaction. The visualization of such interactions enables the inference of possible antibacterial mechanisms. Methods and Results: Nanotextures are synthesized on biocompatible polymer microparticles (MPs) via plasma etching. Both conventional and liquid-phase transmission electron microscopy observations suggest that these MPs may cause cell lysis via bacterial binding to a single protrusion of the nanotexture. The bacterium/protrusion interaction locally compromises the cell wall, thus causing bacterial death. This study suggests that local mechanical damage and leakage of the cytosol kill the bacteria first, with subsequent degradation of the cell envelope. Conclusion: Nanoscale surface roughness may act via a penetrative bactericidal mechanism. This insight suggests that future research may focus on optimizing bacterial binding to individual nanoscale projections in addition to stretching bacteria between nanopillars. Further, antibacterial nanotextures may find use in novel applications employing particles in addition to nanotextures on fibers or films.

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A Review on Macroscale and Microscale Cell Lysis Methods

Cited by 326 publications

References 106 publications

Characterization of Bacterial Lysates by Use of Matrix-Assisted Laser Desorptionâ€“ Ionization Time of Flight Mass Spectrometry Fingerprinting

Characterization of Bacterial Lysates by Use of Matrix-Assisted Laser Desorptionâ€“ Ionization Time of Flight Mass Spectrometry Fingerprinting

Comparative Evaluation of Soxhlet and Ultrasonics on the Structural Morphology and Extraction of Bioactive Compounds of Lemon (Citrus limon L.) Peel

<p>Correlative ex situ and Liquid-Cell TEM Observation of Bacterial Cell Membrane Damage Induced by Rough Surface Topology</p>

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