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Recovering What Matters: High Protein Recovery after Endotoxin Removal from LPS-Contaminated Formulations Using Novel Anti-Lipid A Antibody Microparticle Conjugates. | LitMetric

AI Article Synopsis

  • Endotoxins (lipopolysaccharides) from Gram-negative bacteria can trigger severe immune responses, necessitating strict limits on their presence in injectable drugs, leading to the need for effective removal systems.
  • Most current endotoxin removal methods are non-specific, which can accidentally remove important proteins from drug formulations, causing issues in efficacy.
  • This study introduces anti-Lipid A antibody-conjugated microparticles that selectively target and remove endotoxins without losing essential proteins, demonstrating higher recovery rates in various formulations compared to traditional non-specific methods.

Article Abstract

Endotoxins or lipopolysaccharides (LPS), found in the outer membrane of Gram-negative bacterial cell walls, can stimulate the human innate immune system, leading to life-threatening symptoms. Therefore, regulatory limits for endotoxin content apply to injectable pharmaceuticals, and excess LPS must be removed before commercialization. The majority of available endotoxin removal systems are based on the non-specific adsorption of LPS to charged and/or hydrophobic surfaces. Albeit effective to remove endotoxins, the lack of specificity can result in the unwanted loss of essential proteins from the pharmaceutical formulation. In this work, we developed microparticles conjugated to anti-Lipid A antibodies for selective endotoxin removal. Anti-Lipid A particles were characterized using flow cytometry and microscopy techniques. These particles exhibited a depletion capacity > 6 ×10 endotoxin units/mg particles from water, as determined with two independent methods (Limulus Amebocyte Lysate test and nanoparticle tracking analysis). Additionally, we compared these particles with a non-specific endotoxin removal system in a series of formulations of increasing complexity: bovine serum albumin in water < insulin in buffer < birch pollen extracts. We demonstrated that the specific anti-Lipid A particles show a higher protein recovery without compromising their endotoxin removal capacity. Consequently, we believe that the specificity layer integrated by the anti-Lipid A antibody could be advantageous to enhance product yield.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10531372PMC
http://dx.doi.org/10.3390/ijms241813971DOI Listing

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