Trisacryl

DEAF Sepharose Fast Flow LC DEAF Sepharose Fast Flow HC DEAF. Spherodex M DEAF. Spherosil M DEAF. Trisacryl Plus M Toyopead DEAE-650 (M) Fractogel EMD DMAE-650 (M) Fractogel EMD DEAE-650 (M) Q Sepharose Fast Flow QMA Spherodex M QMA Spherosil M QMA Trisacryl Plus M Toyopead QAE-550 C SP Sepharose Fast Flow SP Sepharose High Performance SP Sepharose Big Bead SP Trisacryl Plus M Toyopead SP-650 M Fractogel EMD SO -650 M... 

Sephadex G25 Medium Sephadex G75 Trisacryl Plus GF 03 M Trisacryl Plus GF 10 M Trisacryl Plus GF 20 M Sephacryl S-IOOHR Sephacryl S-200HR Toyopead HW-50F Toyopead HW-55F... 

Load diluted supernatant onto a column of Pharmacia DEAE Trisacryl. 

Xylanase was obtained from filtered fermentation broths of Chainia NCL-82-5-1 (6). It was precipitated by the addition of three volumes of 0-4°C ethanol, separated from the supernatant by centrifugation, and lyophilized. Four column chromatographic steps, DEAE-cellulose anion exchange, Fractogel TSK DEAE-650S anion exchange, Sephadex G-50-50 gel permeation, and CM-Trisacryl cation exchange, were used for pu ication. 

Xylanase II was subjected to cation exchange chromatography with a 20-mm i.d., 700-mm long CM-Trisacryl column eluted with 0.8 mL/min of 0.025M sodium acetate buffer at pH 4.5, with a 0-0.2M sodium chloride gradient. Only one protein peak eluted from the column it contained all the Xylanase n activity. This material appeared... 

In various runs, specific activity went from an average of 2.75 lU/mg protein (based on the Bio-Rad protein assay with bovine serum albumin standard) for Ae centrifuged crude preparation after alcohol precipitation, to 8 lU/mg after DEAE-cellulose chromatography, to 100 lU/mg after DEAE-Fractogel chromatography, to 1000 RJ/mg after Sephadex G-50 chromatography, and to 3200 lU/mg after CM-Trisacryl chromatography. 

Figure 3.29.A shows a flow-cell of 20 iL inner volume used to hold immobilized anti-mouse IgG bound to a rigid beaded support (activated Pierce trisacryl GF-2000). The cell was used to develop a two-site immunoassay for mouse IgG by consecutive injection of the sample, acridinium ester-labelled antibody and alkaline hydrogen peroxide to initiate the chemiluminescence, which started the reaction sequence shown in Fig. 3.29.B. Regenerating the sensor entailed subsequent injection of an acid solution, which resulted in a determination time of ca. 12 min (this varied as a fimction of the flow-rate used, which also determined the detection limit achieved, viz. 50 amol for an overall analysis time of 18 min) [218]. The sensor was used for at least one week with an inter-assay RSD of 5.9%. Attempts at automating the hydrodynamic system for use in routine analyses are currently under way.

Information in diis table was obtained from Ref. [26]. and has been updated to indicate diat Amersham is now part of GE Healdicare. Most of die fisted supports are available in preactivated forms. Listed supports that are only available widi specific hgands attached are Affi-Gel 601, Affi-Prep, die Mimetic series, Sepharose Big Beads, SigmaChrom AF, and Trisacryl. 

Ultrogel, Magnogel, and Trisacryl Practical guide in affinity chromatoga-phy and related techniques, 2nd ed. (1983) Reactifs IBP, VUleneuve-la-Garenne... 

Figure 4.3—Copolymerisation of two unsaturated monomers (acid and trihydroxyamide). Example of the structure obtained (CM-TRISACRYL M K of IBP France).

Agarose gels are excellent at the laboratory level, but their high cost precludes their use in Industry. The cheaper Trisacryl, an all-synthetic polymer having similar properties, has gained wide acceptance for technical applications.14... 

The bacterial ACV synthetase from S. clavuligerus had been investigated by Kadima and co-workers [88] with the aim of producing the tripeptide. The enzyme activity was stabilized by glycerol, dithiothreitol, Mg2+, and the substrate amino acids, with a half-life of 6 days at 4°C. Immobilization of the synthetase to DEAE-Trisacryl or aminopropyl-glass resulted in a drastic loss of activity, estimated as 5 pmol ACV/min mg immobilized enzyme [28]. In an ultrafiltration cell at room temperature and a nitrogen atmosphere, 45-65% activity was retained after 5 uses during a 24-hr period. Extrapolation of the data indicated that 1.3 g of partially purified synthetase would produce 411 mg of ACV in such a period, which corresponds to a specific activity of 600 pmol/min mg. 

FIGURE 39 Operating regions based on the product yield and processing rate for a human serum albumin (HAS)-DEAE Trisacryl M ion-exchange chromatographic system. The open circles are the maximum production rates (0.87 and 1.46 mg/mL- min) for the 100 and 99% product yield curves derived from the tanks-in-series sectional model approach. Data from Mao, Q. M. and Hearn, M. T. W 1996. Biotechnol. Bioeng., 52, 202. Reprinted by permission of Wiley-Liss, Inc., a subsidiary of John Wiley Sons, Inc. 

FIGURE 8 Separation of rabbit polyclonal antibodies by ion-exchange chromatography on DEAE Trisacryl M. Column dimensions 16 mm i.d.X 100 mm initial buffer 50 mM Tris-HCI, 0.035 M sodium chloride, pH 8.8 load 5 mL of rabbit serum previously precipitated with ammonium sulfate at 50% saturation and redissolved in column buffer flow rate 50 mL/hr elution of adsorbed protein performed using I M sodium chloride solution in the initial Tris buffer. The first peak represents IgG the second peak is composed of all other serum proteins precipitated by ammonium sulfate. The straight line is absorbance at 280 nm, and the broken line represents the variation of ionic strength of the buffer. The purity of IgG estimated by gel electrophoresis was over 98% and the calculated yield was over 90%. 

The most commonly used biopolymers, such as agarose, contain alcoholic hydroxyl groups which can be activated with cyanogen bromide however, better methods have recently been developed including activation with sulfonyl chlorides (17), 2-fluoro-l-methylpyridinium toluene sulfonate (FMP) (10), and chlorocarbonates (18). The first two are commercially available as activated supports tresyl-activated Sepharose (Pharmacia) and FMP-Trisacryl (BioProbe International). The newer methods yield more stable bonds, which preclude leaching of the enzyme from the matrix. Most of these activated supports are too expensive for commercial use in a large process bioreactor however, they may be extremely useful for preparing analytical bioreactors. 

Figure 4.5 Copolymerization of two monoethylenic monomers (an acid and a trihydroxyamide). Example of the structure obtained (CM-TRISACRYL M of IBF-France). Arising from a weak acid the resultant phase wih be unusable at acid pH, as it will no longer be in its ionized form.

Trisacryl gelatin microspheres (Embosphere) calibrated, flexible, non-aggregating... 

Chua GC, Wilsher M, Young MP, Manyonda I, Morgan R, Belli AM (2005) Comparison of particle penetration with non-spherical polyvinyl alcohol versus trisacryl gelatin microspheres in women undergoing premyomectomy uterine artery embolization. Clin Radiol 60 116-122... 

Fig. 2.5a,b. Trisacryl gelatin microspheres or embospheres (courtesy of Biosphere Medical), a Trisacryl gelatin microspheres in suspension, b Microscopic images demonstrating the spheres... 

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