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Related Concept Videos

Bacterial Toxins01:12

Bacterial Toxins

Bacterial toxins are sophisticated virulence factors that enable pathogenic bacteria to interact with, invade, and damage host tissues. These toxins fall broadly into two types: protein exotoxins, which are secreted into the environment and target specific host receptors, and lipopolysaccharide endotoxins, which are structural components of the bacterial outer membrane released primarily during bacterial lysis or membrane shedding. Exotoxins generally act more selectively, binding to cell...
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Botulism is a life-threatening neuroparalytic condition caused by botulinum neurotoxin, which is produced by the bacterium Clostridium botulinum, a Gram-positive, spore-forming, obligate anaerobe.In adults, the toxin enters the body in different ways: in foodborne botulism, the preformed toxin is absorbed in the intestine. In wound botulism, spores grow in injured tissue and release the toxin into the blood. Infant botulism differs mechanistically from adult forms. In infants, botulism commonly...
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Diphtheria is an acute, toxin-mediated infectious disease that primarily affects the upper respiratory tract. It is caused by Corynebacterium diphtheriae, a Gram-positive, pleomorphic rod that lacks spore-forming capability and exhibits a characteristic club-shaped morphology under microscopic examination. While C. diphtheriae can asymptomatically colonize mucosal surfaces, clinical disease manifests only when the bacterial strain is lysogenized by a specific β-corynephage. This phage...
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Microorganisms rely on proteins as an essential carbon and energy source, particularly in environments with limited polysaccharides or lipids. However, proteins are too large to cross the plasma membrane unaided, necessitating enzymatic degradation. Microbes secrete extracellular proteases and peptidases that hydrolyze proteins into peptides, which can then be transported across the membrane. Once inside the cell, intracellular proteases degrade these peptides into free amino acids, which...
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Tetanus is a life-threatening neurological disorder characterized by persistent muscle contractions and spastic paralysis. It is caused by Clostridium tetani, a motile, Gram-positive, rod-shaped, obligate anaerobe. These bacteria produce terminal endospores, giving them a distinctive “lollipop” or “tennis-racket” appearance. They thrive in anaerobic environments, such as those found in deep puncture wounds.Once introduced into the body, the spores germinate into vegetative cells. These cells...
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Anthrax is a zoonotic disease caused by Bacillus anthracis, a Gram-positive, spore-forming bacterium. It primarily affects herbivorous animals but can be transmitted to humans through skin contact, ingestion, or inhalation of spores.Cutaneous anthrax, the most common form, typically results from direct contact with bacterial spores through skin abrasions and is generally less severe. Gastrointestinal anthrax results from eating undercooked or contaminated meat. It affects the mouth, throat, or...

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Detection of Toxin Translocation into the Host Cytosol by Surface Plasmon Resonance
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Published on: January 3, 2012

Endotoxin deactivation by transient acidification.

Melina M Ribeiro1, Xiumin Xu, Dagmar Klein

  • 1Diabetes Research Institute, University of Miami Leonard Miller School of Medicine, Miami, FL 33136, USA.

Cell Transplantation
|April 24, 2010
PubMed
Summary

This study introduces a simple, cost-effective method to reduce endotoxin contamination in recombinant proteins. Transient exposure to acidic conditions effectively deactivates endotoxins, ensuring safer therapeutic protein applications.

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Area of Science:

  • Biotechnology
  • Protein Chemistry
  • Immunology

Background:

  • Recombinant proteins are vital research and therapeutic tools.
  • Bacterial propagation contaminates proteins with endotoxins, necessitating removal for safe use.
  • Endotoxins interfere with assays and cause pathological effects in vivo.

Purpose of the Study:

  • To develop a simple, efficient method for reducing endotoxin activity in recombinant proteins.
  • To validate the method's efficacy across various recombinant proteins, including therapeutic antibodies.
  • To assess the impact of the method on protein recovery and biological activity.

Main Methods:

  • A one-step protocol involving transient exposure of recombinant proteins to acidic conditions (pH below isoelectric point).
  • In vitro endotoxin assessment using Limulus Amebocyte Lysate (LAL) assay.
  • In vivo assessment using pyrogenicity testing.
  • Verification of protein activity through binding assays (e.g., anti-CD154 antibody).

Main Results:

  • Over 99% endotoxin deactivation achieved for all tested recombinant proteins.
  • Protein recovery ranged from 46.8% to 72.9%.
  • Biological activity of tested proteins, including anti-CD154 antibody, remained unchanged.
  • Method effectiveness was not significantly impacted by urea.

Conclusions:

  • Acidic treatment offers a simple, cost-efficient solution for reducing endotoxin levels in recombinant proteins and antibodies.
  • This method preserves protein integrity and biological function, facilitating safer therapeutic applications.
  • The protocol eliminates the need for additional purification steps, streamlining protein preparation.