Chromatographic removal of endotoxin from protein solutions by polymer particles

Hirayama, Chuichi; Sakata, Masayo

doi:10.1016/s1570-0232(02)00430-0

Cited by 114 publications

(85 citation statements)

References 49 publications

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“…These results led to the development of a ''flash'' heating step for the CO complex followed by a Q Sepharose FF column, both of which remove large amounts of weakly bound LPS, free porphyrin, and other contaminants and decrease the endotoxin levels to £ 2 EU/g (109). Polymyxin B agarose, diethylaminoethyl (DEAE) cellulose, or e-poly-L-lysine cellulose can be used to safely and efficiently remove any remaining LPS as long as breakdown of the column matrix does not occur (61). These techniques led to the use of the first (rHb1.1) and second (rHb2.0) products in humans where, as described in previous sections, the observed side effects were much less severe and appeared to be due to NO scavenging (Fig.…”

Section: Endotoxin Removalmentioning

confidence: 99%

Development of Recombinant Hemoglobin-Based Oxygen Carriers

Varnado

Mollan

Birukou

et al. 2013

Antioxidants & Redox Signaling

102

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Significance: The worldwide blood shortage has generated a significant demand for alternatives to whole blood and packed red blood cells for use in transfusion therapy. One such alternative involves the use of acellular recombinant hemoglobin (Hb) as an oxygen carrier. Recent Advances: Large amounts of recombinant human Hb can be expressed and purified from transgenic Escherichia coli. The physiological suitability of this material can be enhanced using protein-engineering strategies to address specific efficacy and toxicity issues. Mutagenesis of Hb can (i) adjust dioxygen affinity over a 100-fold range, (ii) reduce nitric oxide (NO) scavenging over 30-fold without compromising dioxygen binding, (iii) slow the rate of autooxidation, (iv) slow the rate of hemin loss, (v) impede subunit dissociation, and (vi) diminish irreversible subunit denaturation. Recombinant Hb production is potentially unlimited and readily subjected to current good manufacturing practices, but may be restricted by cost. Acellular Hb-based O 2 carriers have superior shelf-life compared to red blood cells, are universally compatible, and provide an alternative for patients for whom no other alternative blood products are available or acceptable. Critical Issues: Remaining objectives include increasing Hb stability, mitigating iron-catalyzed and iron-centered oxidative reactivity, lowering the rate of hemin loss, and lowering the costs of expression and purification. Although many mutations and chemical modifications have been proposed to address these issues, the precise ensemble of mutations has not yet been identified. Future Directions: Future studies are aimed at selecting various combinations of mutations that can reduce NO scavenging, autooxidation, oxidative degradation, and denaturation without compromising O 2 delivery, and then investigating their suitability and safety in vivo. Antioxid. Redox Signal. 18, 2314-2328.

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Section: Endotoxin Removalmentioning

confidence: 99%

Development of Recombinant Hemoglobin-Based Oxygen Carriers

Varnado

Mollan

Birukou

et al. 2013

Antioxidants & Redox Signaling

102

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“…Poly(ε-lysine)-immobilized cellulose beads (PL-Cellufine) [8][9][10] was obtained from Chisso Co. Ltd. (Tokyo, Japan). PL (Mw, 4500; pKa, 7.6; Chisso) was produced by Streptomyces albulus.…”

Section: Reagents and Materialsmentioning

confidence: 99%

“…We previously reported 8,9 that the adsorption activity of PL-Cellufine for bio-products is due to the simultaneous effects of their cationic and hydrophobic or other properties. PL-Cellufine and DEAE-Sepharose, being cationic in nature, can adsorb LPS and acidic proteins mainly by ionic interaction, because the charge of LPS (pK1 = 1.3) 13 and acidic proteins (ovalbumin (pI = 4.6), BSA (pI = 4.9)) are anionic at neutral pH.…”

Section: Selective Adsorption Of Lps Onto Pl-cellufinementioning

confidence: 99%

Limulus Amebocyte Lysate Assay for Endotoxins by an Adsorption Method with Polycation-immobilized Cellulose Beads

et al. 2010

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introductionThe pharmaceutical products for parenteral administration must be tested for contamination of bacterial endotoxins (LPSs 1 ) because of their potent biologic activities, which can cause pyrogenic and shock reactions. LPS, a constituent of the cell walls of Gram-negative bacteria, is a potential contaminant of protein solutions originating from biologic products. There are two methods for determining LPS. One is the rabbit pyrogen test; the other is the limulus test using Limulus amebocyte lysate (LAL).The gel-clot, 2 turbidimetric, 3 and chromogenic 4 testing methods, methods previously approved by the USA Pharmacopoeia in 1981, 1985, and 1985, respectively, are typically employed in the LAL test. The advantages of the LAL test over the rabbit pyrogen test are higher sensitivity, lower cost, and higher reproducibility. The LAL test is inhibited or enhanced by antibiotics, hormones, heavy metals, amino acids, alkaloids, plasma proteins, enzymes, and electrolytes in the sample solution.5 Dilution, heating and dialysis have been used to overcome these problems, but the results have been unsatisfactory. Minobe et al. attempted to develop a new specific assay method for LPSs using histidine-immobilized Sepharose as an adsorbent.6,7 LPSs adsorbed by the adsorbent were separated from the compounds (e.g., amino acids, enzymes) in solution by centrifugation, and the adsorbed LPSs can react directly with the LAL. The great disadvantage of this type of adsorbent is the low adsorption activity of the LPSs in physiologic solution (neutral pH and ionic strength (μ) = 0.1 -0.2).In this study, we developed a novel assay of LPSs with a new adsorption method using a turbidimetric time assay system. We previously found that poly(ε-lysine)-immobilized cellulose beads (PL-Cellufine) can selectively adsorb LPSs in bioproducts over a wide range of ionic strengths (μ) from 0.05 -0.8 8-10 at neutral pH. On the other hand, a simple turbidimetric time assay system has been developed for the quantitative assaying of LPS; an automated apparatus is available commercially. 11In this paper, we describe the LPS-adsorbing activities of PL-Cellufine. Then, using the turbidimetric time assay, we describe the optimum conditions for the LPS assay using the PL-Cellufine as adsorbent. We also compare the apparent quantity of LPS provided by the adsorption method with the apparent quantity of LPS provided by a normal method. experimental Reagents and materialsPurified LPS (Escherichia coli UKT-B and O111:B4) and Limulus ES-II test Wako (Limulus amebocyte lysate) were purchased from Wako Pure Chemicals (Osaka, Japan). Poly(ε-lysine)-immobilized cellulose beads (PL-Cellufine) [8][9][10] was obtained from Chisso Co. Ltd. (Tokyo, Japan). PL (Mw, 4500; pKa, 7.6; Chisso) was produced by Streptomyces albulus. 12 DEAE-Sepharose CL-6B was purchased from GE Healthcare Bio-Sciences Corp. (USA). Ovalbumin (from egg), bovine serum albumin (BSA), γ-globulin (from human serum), lysozyme (from horse heart), and vitamin K3 were purchased from Wako. Myoglobi...

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“…The purity of the protein preparations was checked by SDS-PAGE as described previously (19,20). All Gal preparations were checked for endotoxin contamination by Limulus assay (BioWhittaker); any contaminating LPS was removed with the use of polymer particles as described previously (21). The final endotoxin content of Gal used in this study was Ͻ0.008 endotoxin units (equivalent to 0.8 pg of LPS).…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

Galectin-9 Induces Maturation of Human Monocyte-Derived Dendritic Cells

Dai

Nakagawa

Itoh³

et al. 2005

The Journal of Immunology

189

126

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Maturation of dendritic cells (DCs) is critical for initiation of immune responses and is regulated by various stimulatory signals. We assessed the role of galectin (Gal)-9 in DC maturation. Culture of immature DCs with exogenous Gal-9 markedly increased the surface expression of CD40, CD54, CD80, CD83, CD86, and HLA-DR in a dose-dependent manner, although Gal-9 had no or little effect on differentiation of human monocytes into immature DCs. Gal-9-treated DCs secreted IL-12 but not IL-10, and they elicited the production of Th1 cytokines (IFN-γ and IL-2) but not that of the Th2 cytokines (IL-4 and IL-5) by allogeneic CD4+ T cells. These effects of Gal-9 on immature DCs were not essentially dependent on its lectin properties, given that they were inhibited only slightly by lactose. We further found that a Gal-9 mutant that lacks β-galactoside binding activity reproduced the above activities and that an anti-Gal-9 mAb suppressed them. Gal-9 induced phosphorylation of the MAPK p38 and ERK1/2 in DCs, and an inhibitor of p38 signaling, but not inhibitors of signaling by either ERK1/2 or PI3K, blocked Gal-9-induced up-regulation of costimulatory molecule expression and IL-12 production. These findings suggest that Gal-9 plays a role not only in innate immunity but also in acquired immunity by inducing DC maturation and promoting Th1 immune responses.

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Chromatographic removal of endotoxin from protein solutions by polymer particles

Cited by 114 publications

References 49 publications

Development of Recombinant Hemoglobin-Based Oxygen Carriers

Development of Recombinant Hemoglobin-Based Oxygen Carriers

Limulus Amebocyte Lysate Assay for Endotoxins by an Adsorption Method with Polycation-immobilized Cellulose Beads

Galectin-9 Induces Maturation of Human Monocyte-Derived Dendritic Cells

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