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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

鲎试剂 牛血清白蛋白 化学 脂质A 溶解 抗体 微粒 色谱法 流式细胞术 脂多糖 生物化学 生物 分子生物学 免疫学 天体生物学
作者
Cristiane C. De Melo,Alexandra C. Fux,Martin Himly,Neus G. Bastús,Laura Schlahsa,Christiane Siewert,Víctor Puntes,Albert Duschl,Isabel Gessner,Jonathan A. Fauerbach
出处
期刊:International Journal of Molecular Sciences [MDPI AG]
卷期号:24 (18): 13971-13971 被引量:2
标识
DOI:10.3390/ijms241813971
摘要

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 ×103 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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