Cibacron Blue, immobilized

The effects of a washing step on the position and shape of a breakthrough curve and the amount of protein retained in the column are shown in Fig. 37 using hen egg white lysozyme and Cibacron Blue F3GA immobilized Fractosil 1000 system as the exemplar. In Fig. 37a the time-concentration... 

As observed with this HEWL-Cibacron Blue F3GA immobilized Frac-tosil 1000 system, and numerous other cases of polypeptide or protein interaction with HPLC sorbents, the maximum production rate tends to increase with the increase of the terminating effluent concentration. At fluid velocities lower than the optimum velocities, the effect of the terminating effluent concentration, however, becomes less important. The use of a flow rate at the maximum capacity of the pump (or to the pressure limit of the system as is sometimes practiced) will usually lead to an impaired production rate with HP-BAC, HP-BMC, and HP-HIC sorbents.307,422,423 This conclusion has been also supported by other experimental data on large-scale chromatographic purification of proteins with HP-IEX sorbents.368,406,421,424... 

FIGURE 40 Effect of changes in the feed superficial velocity on the experimental breakthrough curve for the adsorption of HEWL to Cibacron Blue F3G-A immobilized onto Fractosil 1000 as a function of time and the amount of protein added to the column. Data adapted from Ref. 436. 

FIGURE 42 Influence of superficial velocity on the DAC and the DAR obtained for the adsorption of (a) HEWL and (b) HSA to Cibacron Blue F3G-A immobilized onto Fractosil 1000. Data from Finette, G. M. S., Mao, Q. M and Hearn, M. T. W., 1998, Biotechnol. Bioeng., 58, 35. Reprinted by permission of Wiley-Liss, Inc., a subsidiary of John Wiley Sons, Inc. 

FIGURE 43 Comparison of the shape and position of the breakthrough curves in terms of time or the amount of protein applied and generated for the corresponding packed and fluidized columns for the adsorption of HEWL (a) and (b), respectively, and HSA (c) and (d), respectively, to Cibacron Blue F3G-A immobilized onto Fractosil 1000. The superficial velocities applied were 0.64 cm/min in (a) and (b) and 3.84 cm / min in (c) and (d), respectively. 

Displacement chromatography has also been carried out in dye affinity chromatographic systems for the purification of lactate dehydrogenase (LDH).59 In that study, polyethyleneimine (PEI) was employed as a displacer on dye affinity matrices prepared by immobilizing Cibacron Blue 3GA or Procion Red HE-3B. 

Immobilied Cibacron Blue 3G-A has been found very efficient for the isolation of human serum albumin interferons produced in tissue cultures phosphogly-cerate kinase from Saccharomyces cerevisiae and alcohol dehydrogenase from equine liver. Immobilized Procion Red HE-3B and Prcxaon Blue MX-4GD have been successfully applied for the isolation of 3-hydroxybutyrate and malate dehydrogenases... 

Gianazza E, Amaud P (1982) Chromatography of plasma proteins on immobilized Cibacron Blue F3-GA. Mechanism of the molecular interaction. Biochem J 203 637-641. 

GlOa. Ghiggeri, G. M., Candiano, G., Delflno, G., and Queirolo, C., Highly selective one-step chromatography of serum and urinary albumin on immobilized Cibacron Blue F3GA. Clin. Chim. Acta 145, 205-211 (1985). 

Ricin, the toxin of Ricinus communis, has been adsorbed on guar gum, a polysaccharide containing D-galactosyl and D-mannosyl residues.The nonbinding toxic A chain is then isolated by reduction of the immobilized toxin with 2-mercaptoethanol, and the B chain is eluted with lactose. A nucleotidebinding domain in the A-chain was demonstrated from its interaction with Cibacron Blue F3GA. 

The structxire of the component proteins of complement have been reviewed. All nine components of the human classical complement pathway bind to immobilized Cibacron Blue F3GA, and are eluted without contamination of substrates by albumin or immunoglobulin The sub-component... 

The PEI-coated membrane can be used as a support for immobilization of the affinity ligand. The Cibacron Blue F3GA dye was chosen as the affinity ligand and immobilized on the PEI coating layer. In the case of the membrane coated with 1% (v/v) PEI coating solution, the amount of dye coupled on the membrane, roughly estimated by the method described by Champluvier (9), was about 1.0 pmol/mL of the porous layer... 

An interesting phenomenon is that the PSU nanofibers become curled after PMAA grafting by Ce(lV) and the protein ligands such as Cibacron blue F3GA (CB) can be covalently immobilized on the PMAA-grafted PSU nanofiber membrane... 

Ma, Z.W., Masaya, K. and Ramakrishna, S. 2006b. Immobilization of Cibacron blue F3GA on eiectrospun polysulphone ultra-fine fiber surfaces towards developing an affinity membrane for albumin adsorption. 

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