A Method for Extracorporeal Heparin Removal from Blood by Affinity Chromatography

Hou, Kenneth C.; Roy, Sujata; Zaniewski, Richard P.; Shumway, Edward

doi:10.1111/j.1525-1594.1990.tb03000.x

Cited by 11 publications

(9 citation statements)

References 18 publications

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“…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. As shown in Figs.…”

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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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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“…Hou et al 14 previously reported that a hydrophobic bond was formed between LPS and the polymeric affinity matrix. It is possible that the hydrophobic chains (the center-to-center distance 0.49 nm) 9 of the LPS monomer are included into the cavities (diameter 0.70 and 0.85 nm, respectively) of β-and γ-CyD polymers beads (Fig.…”

Section: Resultsmentioning

confidence: 99%

Selective Removal of Endotoxin from a DNA Solution by Cross-linked Cyclodextrin Beads

et al. 2011

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“…The material used was cellulose membrane and the preparation of immunoaffinity media and column has been described previously [20]. Two approaches were used for the immobilization of protein A on the acrylic and amino membranes, respectively.…”

Section: Preparation Of Affinity Columnsmentioning

confidence: 99%

“…Those new matrices have the advantages of fast analysis and simple scale-up, but matrices for perfusion chromatography cost much more than membrane matrices. Cellulose membrane has been used for analysis and purification of biopolymers for its good biological compatibility, high efficiency and low cost [19,20]. We have synthesized a modified cellulose membrane matrix and protein A and human immunoglobulin G (IgG) were immobilized on such membrane supports for the fast analysis of human IgG and goat polyclonal antibodies to human IgG with a column: 20 x 4mm I.D.…”

Section: Introductionmentioning

confidence: 99%

Membrane affinity chromatography for analysis and purification of biopolymers

Zhou

Zou

et al. 1999

Chromatographia

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KeyWordsColumn liquid chromatography Membrane stationary phase Immunoaffinity analysis Affinity efficiency Adsorption capacity SummaryAffinity columns suitable for HPLC were prepared by immobilization of various ligands of protein A, human IgG, human IgM and pectinase on GMA modified cellulose membrane. The adsorption capacity, affinity efficiency and activity recovery of various IgGs on these affinity columns were measured. It was observed that the length of the coupling arm plays a very important role in affinity efficiency, and the effect of eluent flow-rate on adsorption capacity was very small. The protein A column was exploited for the process monitoring of dog IgG in clinical experiments on immuno-adsorption therapy. A pectinase column was used for the determination of polygalacturonase inhibiting proteins first purified on a hydroxyapatite column. It took only about 2.5 min for analysis at a flow-rate of 1.0 mL min -1. The high speed analysis of biopolymers could be performed at a flow rate of 6.0 1 mL rain-within 15 s. Membrane affinity chromatography gives good reproducibility, high efficiency, low column-pressure and is rapid. It can also be used for micro-scale purification ofbiopolymers.

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A Method for Extracorporeal Heparin Removal from Blood by Affinity Chromatography

Cited by 11 publications

References 18 publications

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

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

Selective Removal of Endotoxin from a DNA Solution by Cross-linked Cyclodextrin Beads

Membrane affinity chromatography for analysis and purification of biopolymers

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