Membrane adsorber for endotoxin removal

ABSTRACT The surface of flat-sheet nylon membranes was modified using bisoxirane as the spacer and polyvinyl alcohol as the coating polymer. The amino acid histidine was explored as a ligand for endotoxins, aiming at its application for endotoxin removal from aqueous solutions. Characterization of the membrane adsorber, analysis of the depyrogenation procedures and the evaluation of endotoxin removal efficiency in static mode are discussed. Ligand density of the membranes was around 7 mg/g dry membrane, allowing removal of up to 65% of the endotoxins. The performance of the membrane adsorber prepared using nylon coated with polyvinyl alcohol and containing histidine as the ligand proved superior to other membrane adsorbers reported in the literature. The lack of endotoxin adsorption on nylon membranes without histidine confirmed that endotoxin removal was due to the presence of the ligand at the membrane surface. Modified membranes were highly stable, exhibiting a lifespan of approximately thirty months

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Evaluation Of A Chitosan Membrane For Removal Of Endotoxin From Human Igg Solutions

Cross-linked chitosan membranes - prepared with silica particles as porogen agent - were evaluated for the removal of endotoxin (ET) from human immunoglobulin G (IgG) preparations. The effects of solution conditions on the efficiency of ET removal and IgG recovery were studied. The depyrogenation studies were performed with Tris-HCl at pH 7.0 due to low IgG adsorption and significant degree of protonation (0.50) of the membrane in this buffer. Adsorption ET data were analyzed using the Langmuir model (maximum binding capacity and dissociation constant were 280 μg/mL and 4.0 × 10-11 mol/L, respectively). A high ET clearance (96%) and IgG recovery (99%) were obtained with ET and IgG initial concentration of 116.4 EU/mL and 1.0 mg/mL, respectively. Therefore, these results assure the potential of using chitosan membrane filtration for ET removal in the downstream processing of IgG solutions. © 2006 Elsevier Ltd. 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Adsorption Of Human Immunoglobuling Onto Ethacrylate And Histidine-linked Methacrylate

The adsorption of human IgG onto GMA (a semirigid methacrylate-based chromatography matrix) and His-GMA adsorbents was studied by chromatography and batch equilibrium binding analysis. IgG molecules adsorbed onto GMA gel by nonspecific hydrophobic interactions and the specificities were similar for both adsorbents. Adsorption data were analyzed using three isotherm models, namely the Langmuir, Freundlich and Langmuir-Freundlich models, and the adsorption parameters were computed. The experimental isotherms were best described by a combined Langmuir-Freundlich model, which indicated the presence of unequal binding sites on both adsorbents and/or positive cooperativity in the binding of the IgG molecules.203251262Adamson, A.W., (1990) Physical Chemistry of Surfaces, Fifth Edition, , John Wiley and Sons, Inc., New YorkAndrade, J.D., (1985) Surface and Interfacial Aspects of Biomedical Polymers, 2, pp. 1-80. , in J.D. 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Evaluation Of Immobilized Metal Membrane Affinity Chromatography For Purification Of An Immunoglobulin G1 Monoclonal Antibody

The large scale production of monoclonal antibodies (McAbs) has gaining increased relevance with the development of the hybridoma cell culture in bioreactors creating a need for specific efficient bioseparation techniques. Conventional fixed bead affinity adsorption commonly applied for McAbs purification has the drawback of low flow rates and colmatage. We developed and evaluated a immobilized metal affinity chromatographies (IMAC) affinity membrane for the purification of anti-TNP IgG1 mouse McAbs. We immobilized metal ions on a poly(ethylene vinyl alcohol) hollow fiber membrane (Me 2+-IDA-PEVA) and applied it for the purification of this McAbs from cell culture supernatant after precipitation with 50% saturation of ammonium sulphate. The purity of IgG1 in the eluate fractions was high when eluted from Zn2+ complex. The anti-TNP antibody could be eluted under conditions causing no loss of antigen binding capacity. The purification procedure can be considered as an alternative to the biospecific adsorbent commonly applied for mouse IgG1 purification, the protein G-Sepharose. © 2004 Elsevier B.V. All rights reserved.8161-2259268Huse, K., Böhme, H.-J., Scholz, G.H., (2002) J. Biochem. Biophys. Methods, 51, p. 217Goding, J.W., (1995) Monoclonal Antibodies: Principles and Practice, , New YorkBoden, V., Winzerling, J.J., Vijayalakshmi, M., Porath, J., (1995) J. Immunol. Methods, 181, p. 225Verdoliva, A., Pannone, F., Rossi, M., Catello, S., Manfredi, V., (2002) J. Immunol. Methods, 271, p. 77Tishchenko, G., Hodorvá, B., Šimůnek, J., Bleha, M., (2003) J. Chromatogr. a, 983, p. 125El-Kak, A., Vijayalakshmi, M.A., (1991) J. Chromatogr., 570, p. 29Bueno, S.M.A., Haupt, K., Vijayalakshmi, M.A., (1995) J. Chromatogr. B, 667, p. 57Tishchenko, G., Dybal, J., Mészárosová, K., Sedláková, Z., Bleha, M., (2002) J. Chromatogr. a, 954, p. 115Gaberc-Porekar, V., Menart, V., (2001) J. Biochem. Biophys. Methods, 49, p. 335Porath, J., Olin, B., (1983) Biochemistry, 22, p. 1621Hale, J.E., Beidler, D.E., (1994) Anal. Biochem., 222, p. 29Vançan, S., Miranda, E.A., Bueno, S.M.A., (2002) Process Biochem., 37, p. 573Mészárosová, K., Tishchenko, G., Bouchal, K., Bleha, M., (2003) React. Funct. Polym., 56, p. 27Klein, E., (1991) Affinity Membranes. Their Chemistry and Performance in Adsorptive Separation Processes, 5. , John Wiley and Sons IncHari, P.R., Paul, W., Sharma, C.P., (2000) J. Biomed. Mater. Res., 50, p. 110Castilho, L.R., Anspach, F.B., Deckwer, W.-D., (2002) J. Membr. Sci., 207, p. 253Léo, P., Ucelli, P., Augusto, E.F.P., Oliveira, M.S., Tamashiro, W.M.S.C., (2000) Hybridoma, 19, p. 473Belew, M., Porath, J., (1990) J. Chromatogr., 516, p. 333Bueno, S.M.A., Legallais, C., Haupt, K., Vijayalakshmi, M.A., (1996) J. Membr. Sci., 117, p. 45Bradford, M.M., (1976) Anal. Biochem., 72, p. 248Laemmli, U.K., (1970) Nature, 227, p. 680Morrissey, J.H., (1981) Anal. Biochem., 117, p. 307Beitle, R.R., Ataai, M.M., (1993) Biotechnol. Progr., 9, p. 64Berna, P.P., Mrabet, N.T., Vanbeeumen, J., Devreese, B., Porath, J., Vijayalakshmi, M.A., (1997) Biochemistry, 36, p. 6896Adamson, A.W., (1990) Physical Chemistry of Surfaces, , fifth ed. John Wiley and Sons Inc. New YorkAndrade, J.D., (1985) Surface and Interfacial Aspects of Biomedical Polymers, , J.D. Andrade Plenum Press New YorkJiang, W., Hearn, M.T.W., (1996) Anal. Biochem., 242, p. 45Sharma, S., Agarwal, G.P., (2001) Anal. Biochem., 288, p. 126Quiñones, I., Guiochon, G., (1998) J. Chromatogr. a, 796, p. 15Vijayalakshmi, M.A., (1989) Trends Biotechnol., 7, p. 7

The Effect Of Nacl On The Adsorption Of Human Igg Onto Cm-asp-peva Hollow Fiber Membrane-immobilized Nickel And Cobalt Metal Ions

Over the past decade, immobilized metal-affinity adsorbents have attracted increasing interest for purification of natural and recombinant immunoglobulin G (IgG). In this work, nickel and cobalt metal ions complexed with CM-Asp (carboxymethylaspartate) immobilized on poly(ethylenevinyl alcohol) (PEVA) hollow fiber membranes were evaluated for purification of human IgG from serum. The buffer system and NaCl had important effects on human serum protein adsorption in both adsorbents. Efficient purification of IgG was accomplished in sodium phosphate buffer without NaCl at pH 7.0. Under this condition, the electrostatic interactions are important for adsorption. The Ni(II)-CM-Asp-PEVA had a protein adsorption capacity of 17.5 mg of IgG mL-1 fiber when human serum diluted was loaded in crossflow filtration mode and the eluted IgG had a purity of 82.6 % (based on total protein and IgG, IgM, HSA, and Trf nephelometric analysis). Fitting the experimental IgG adsorption data to the Langmuir and Langmuir-Freundlich models showed that Ni(II)-CM-Asp and Co(II)-CM-Asp had Langmuirean and non-Langmuirean behavior, respectively, with positive cooperativity for IgG-Co(II)-CM-Asp binding, probably due to multipoint interactions (n = 2.12 ± 0.31). Thus, these membranes can be considered as alternative adsorbents for the purification or depletion of IgG from human serum. © 2014 Springer Science+Business Media New York.2005/06/15677688Altintas, E.B., Tuzmen, N., Uzun, L., Denizli, A., Immobilized metal affinity adsorption for antibody depletion from human serum with monosize beads (2007) Ind. Eng. Chem. Res., 46, pp. 7802-7810. , 10.1021/ie061164cAquino, L.C.L., Sousa, H.R.T., Miranda, E.A., Vilela, L., Bueno, S.M.A., Evaluation of IDA-PEVA hollow fiber membrane metal ion affinity chromatography for purification of a histidine-tagged human proinsulin (2006) J. Chromatogr. 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Endotoxin Removal From Solutions Of F(ab′)2 Fragments Of Equine Antibodies Against Snake Venom Using Macroporous Chitosan Membrane

The adsorptive filtration technique using chitosan membranes was used to remove endotoxin (ET) from buffers and bothrops antivenom serum (F(ab′)2 fragments of equine antibodies). Macroporous chitosan membrane was prepared from chitosan using silica particles as porogen agent. The effects of solution conditions on the efficiency of endotoxin removal and protein recovery were studied. The ET concentrations were reduced to 0.58 and 71.6EU/ml, respectively, when Tris-HCl pH 7.0 buffer with 18.6 and 8092.5EU/ml was passed through the chitosan membrane. This buffer was used to evaluate ET removal in the presence of the equine F(ab′)2 fragments. The results showed that the presence of protein in the feed solution did not influence much the clearance of this membrane. Removal of 97% of the ET and recovery of 94% of the F(ab′)2 were obtained at an initial ET and F(ab′)2 concentration of 974.4EU/ml and 18.6mg/ml, respectively. © 2004 Elsevier B.V. All rights reserved.2341-26773Petsch, D., Anspach, F.B., Endotoxin removal from protein solutions (2000) J. Biotechnol., 76, p. 97Sharma, S.K., Endotoxin detection and elimination in biotechnology (1986) Biotechnol. Appl. Biochem., 8, p. 5Li, L., Luo, R.G., Quantitative determination of Ca2+ effects on endotoxin removal and protein yield in a two-stage ultrafiltration process (1999) Sep. Sci. Technol., 34, p. 1729Aida, Y., Pabst, M.J., Removal of endotoxin from protein solutions by phase separation using Triton X-114 (1990) J. Immunol. Meth., 132, p. 191Neidhardt, E.A., Luther, M.A., Recny, M.A., Rapid two-step purification process for the preparation of pyrogen-free murine immunoglobulin G1 monoclonal antibodies (1992) J. Chromatogr., 590, p. 255Matsumae, H., Minobe, S., Kindan, K., Watanabe, T., Sato, T., Tosa, T., Specific removal of endotoxin from protein solutions by immobilized histidine (1990) Biotechnol. Appl. Biochem., 12, p. 129Anspach, F.B., Hilbeck, O., Removal of endotoxin by affinity sorbents (1995) J. Chromatogr. a, 711, p. 81Hirayama, C., Sakata, M., Nakamura, M., Ihara, H., Kunitake, M., Todokoro, M., Preparation of poly(ε-lysine) adsorbents and application to selective removal of lipopolysaccharides (1999) J. Chromatogr. B, 721, p. 187Anspach, F.B., Petsch, D., Membrane adsorbers for selective endotoxin removal from protein solutions (2000) Process Biochem., 35, p. 1005Dainippon Pharmaceuticals, An agent for removing nucleic acids or endotoxin and a method for the removal, Eur. Patent 0,240,348 (1987)Kurita Water Ind., Removing nucleic acid and endotoxin from solution using chitosan treatment, Jpn. Patent 07,289,238 (1995)Morimoto, S., Sakata, M., Iwata, T., Esaki, A., Hirayama, C., Preparations and applications of polyethyleneimine-immobilized cellulose fibres for endotoxin removal (1995) Polym. J., 27, p. 831Brandt, S., Goffe, R.A., Kessler, S.B., O'Connor, J.L., Zale, S.E., Membrane-based affinity technology for commercial scale purifications (1988) BioTechnology, 6, p. 779Thömmes, J., Kula, M.-R., Membrane chromatography - An integrative concept in the downstream processing of proteins (1995) Biotech. Progr., 11, p. 357Petsch, D., Beeskow, T.C., Anspach, F.B., Deckwer, W.-D., Membrane adsorbers for selective removal of bacterial endotoxin (1997) J. Chromatogr. B, 693, p. 79Petsch, D., Deckwer, W.D., Anspach, F.B., Legallais, C., Vijayalakshmi, M.A., Endotoxin removal with poly(ethyleneimine)-immobilized adsorbers: Sepharose 4B versus flat sheet and hollow fibre membranes (1998) J. Chromatogr. B, 707, p. 121Legallais, C., Anspach, F.B., Bueno, S.M.A., Haupt, K., Vijayalakshmi, M.A., Strategies for the depyrogenation of contaminated immunoglobulin G solutions by histidine-immobilized hollow fiber membrane (1997) J. Chromatogr. B, 691, p. 33Acconci, C., Legallais, C., Vijayalakshmi, M.A., Bueno, S.M.A., Depyrogenation of snake antivenom serum solutions by hollow fiber membranes filtration pseudobioaffinity (2000) J. Membr. Sci., 173, p. 235Zeng, X., Ruckenstein, E., Control of pore sizes in macroporous chitosan and chitin membranes (1996) Ind. Eng. Chem. Res., 35, p. 4169Zeng, X., Ruckenstein, E., Membrane chromatography: Preparation and applications to protein separation (1999) Biotech. Progr., 15, p. 1003Davidova, V.N., Yermak, I.M., Gorbach, V.I., Krasikova, I.N., Solov'Eva, T.F., Interaction of bacterial endotoxins with chitosan. Effect of endotoxin structure, chitosan molecular mass, and ionic strength of the solution on the formation of the complex (2000) Biochemistry (Moscow), 65, p. 1082Nakata, T., Destruction of challenged endotoxin in a dry heat oven (1994) J. Pharm. Sci. Technol., 48, p. 59Bradford, M.M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding (1976) Anal. Biochem., 72, p. 248Rinaudo, M., Pavlov, G., Desbrières, J., Influence of acetic acid concentration on the solubilization of chitosan (1999) Polymer, 40, p. 702