Endotoxin removal and prevention for pre‐clinical biologics production

London, Anne Serdakowski; Kerins, Brendan; Tschantz, William R.; Mackay, Kasey

doi:10.1002/biot.201200220

Cited by 29 publications

(14 citation statements)

References 26 publications

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“…Previous studies have shown that endotoxin elutes with protein aggregates in gel filtration chromatography; however, it was unknown if endotoxin was directly associated with the aggregates or was simply eluting at the same time, based on size. Endotoxin monomers are composed of a polysaccharide region and a lipid region, and are well known to form supramolecular aggregates in aqueous solutions because of their amphipathic structures .…”

Section: Resultsmentioning

confidence: 99%

Gel filtration chromatography as a method for removing bacterial endotoxin from antibody preparations

London¹,

Mackay²,

Lihon³

et al. 2014

Biotechnology Progress

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The removal of bacterial endotoxins from biological samples is critical to avoid the potentially fatal pyrogenic response possible when introduced to mammalian systems. Endotoxins have a variety of specific characteristics that can be exploited to target their isolation and subsequent removal, but one that has not been extensively characterized is their difference in size from that of monoclonal antibodies. Here, we present a study which utilizes gel filtration chromatography as a method for endotoxin removal from both aggregated and nonaggregated antibody preparations, outlining a mechanistically simple method for removal of this impurity.

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Section: Resultsmentioning

confidence: 99%

Gel filtration chromatography as a method for removing bacterial endotoxin from antibody preparations

London¹,

Mackay²,

Lihon³

et al. 2014

Biotechnology Progress

Self Cite

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show abstract

“…Although the final preparation still contained detectable endotoxin levels, the amounts were close to Good Manufacturing Practice standards. Given that endotoxin limits for protein-based injectables are typically 0.5 EU/mg, an additional endotoxin removal step(s) [32] may be required for use of recombinant apoA-I isolated by the present protocol in humans. Regardless, overall low endotoxin content together with scalability of the two-step process combines to makes this purification scheme an attractive option to produce WT apoA-I for commercial and pharmaceutical applications.…”

Section: Discussionmentioning

confidence: 99%

A facile method for isolation of recombinant human apolipoprotein A-I from E. coli

Ikon¹,

Shearer²,

Liu

et al. 2017

Protein Expression and Purification

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Apolipoprotein (apo) A-I is the major protein component of high-density lipoprotein (HDL) and plays key roles in the Reverse Cholesterol Transport pathway. In the past decade, reconstituted HDL (rHDL) has been employed as a therapeutic agent for treatment of atherosclerosis. The ability of rHDL to promote cholesterol efflux from peripheral cells has been documented to reduce the size of atherosclerotic plaque lesions. However, development of apoA-I rHDL-based therapeutics for human use requires a cost effective process to generate an apoA-I product that meets “Good Manufacturing Practice” standards. Methods available for production and isolation of unmodified recombinant human apoA-I at scale are cumbersome, laborious and complex. To overcome this obstacle, a streamlined two-step procedure has been devised for isolation of recombinant untagged human apoA-I from E. coli that takes advantage of its ability to re-fold to a native conformation following denaturation. Heat treatment of a sonicated E. coli supernatant fraction induced precipitation of a large proportion of host cell proteins (HCP), yielding apoA-I as the major soluble protein. Reversed-phase HPLC of this material permitted recovery of apoA-I largely free of HCP and endotoxin. Purified apoA-I possessed α-helix secondary structure, formed rHDL upon incubation with phospholipid and efficiently promoted cholesterol efflux from cholesterol loaded J774 macrophages.

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“…However, common sterilization procedures are usually not efficient in removing endotoxin contamination in NP samples. Techniques developed to remove endotoxin from samples are usually called ‘depyrogenation’ techniques [40,49]. …”

Section: General Aspects Of Dds Toxicologymentioning

confidence: 99%

Unintended effects of drug carriers: Big issues of small particles

Parhiz

Khoshnejad

Myerson

et al. 2018

Advanced Drug Delivery Reviews

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Humoral and cellular host defense mechanisms including diverse phagocytes, leukocytes, and immune cells have evolved over millions of years to protect the body from microbes and other external and internal threats. These policing forces recognize engineered sub-micron drug delivery systems (DDS) as such a threat, and react accordingly. This leads to impediment of the therapeutic action, extensively studied and discussed in the literature. Here, we focus on side effects of DDS interactions with host defenses. We argue that for nanomedicine to reach its clinical potential, the field must redouble its efforts in understanding the interaction between drug delivery systems and the host defenses, so that we can engineer safer interventions with the greatest potential for clinical success.

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Endotoxin removal and prevention for pre‐clinical biologics production

Cited by 29 publications

References 26 publications

Gel filtration chromatography as a method for removing bacterial endotoxin from antibody preparations

Gel filtration chromatography as a method for removing bacterial endotoxin from antibody preparations

A facile method for isolation of recombinant human apolipoprotein A-I from E. coli

Unintended effects of drug carriers: Big issues of small particles

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