Anionic exchange chromatography

Another example of vims clearance is for IgM human antibodies derived from human B lymphocyte cell lines where the steps are precipitation, size exclusion using nucleases, and anion-exchange chromatography (24). A second sequence consists of cation-exchange, hydroxylapatite, and immunoaffinity chromatographies. Each three-step sequence utilizes steps based on different properties. The first sequence employs solubiUty, size, and anion selectivity the second sequence is based on cation selectivity, adsorption, and selective recognition based on an anti-u chain IgG (24). 

Lipoproteins may denature on heating and if present during pasteurization can result in the formation of haze or turbidity in the final product. This material was removed traditionally by filtration through asbestos (qv) sheets (6) however, health hazards associated with asbestos have led to its replacement by alternative filter materials (23,37,193). These media have been less effective than asbestos and further measures have been required to ensure the visual clarity of albumin products, eg, further filtration developments for Hpid removal (194), preferential denaturation of contaminants using in-process heat treatment, and anion-exchange chromatography (49). 

Di- and tri-carboxylic acids. Resolution by anion-exchange chromatography. [Bengtsson and Samuelson Anal Chim Acta 44 217b 1969.]... 

Lipoteichoic acids (from gram-positive bacteria) [56411-57-5J. Extracted by hot phenol/water from disrupted cells. Nucleic acids that were also extracted were removed by treatment with nucleases. Nucleic resistant acids, proteins, polysaccharides and teichoic acids were separated from lipoteichoic acids by anion-exchange chromatography on DEAE-Sephacel or by hydrophobic interaction on octyl-Sepharose [Fischer et al. Ear J Biochem 133 523 1983]. 

Nicotinamide adenine dinucleotide phosphate (NADP, TPN) [53-59-8] M 743.4, pK] 1.1 (PO4H2), pK 4.0 (adenine NH ), pKa 6.1 (P04 ). Purified by anion-exchange chromatography in much the same way as for NAD [Dalziel and Dickinson Biochem J 95 311 7965 Biochemical Preparations 11 87 7966]. Finally it is purified by dissolving in H2O and precipitating with 4 volumes of Me2CO and dried in... 

Xylanase (from Streptomyces lividans) [37278-89-0] Mr 43,000 [EC 3.2.1.8]. Purified by anion-exchange chromatography on an Accell QMA column and finally by HPLC using a ProteinPak DEAE 5PW anion-exchange column. Solutions were stored frozen at -70°. [Morosoli et al. Biochem J 239 587 79S6 Wong et al. Microbiol Rev 52 305 1988.]... 

Dried shrimp was ground, defatted with benzene, and then extracted with cold water. The luciferase extracted was purified first by a batch adsorption onto DEAE cellulose (elution with 0.4 M NaCl), followed by gel filtration on a column of Sephadex G-150, anion-exchange chromatography on a column of DEAE-cellulose (gradient elution 0.05-0.5 M NaCl), and gel filtration on a column of Ultrogel AcA 34. The specific activity of the purified luciferase was 1.7 x 1015 photons s 1 mg-1, and the yield in terms of luciferase activity was about 28%. 

The small jellyfish Phialidium gregarium (diameter 15-20 mm) used to be abundant at Friday Harbor, Washington, in summer and autumn until about 1990. Levine and Ward (1982) isolated and purified a Ca2+-sensitive photoprotein from this jellyfish and named it phialidin. They extracted the photoprotein from whole specimens with an EDTA-containing buffer. The photoprotein extract was precipitated with ammonium sulfate, and purified by the following methods gel-filtration (BioGel P-150, minus 400 mesh), anion-exchange chromatography (DEAE Bio-Gel A), and gel-filtration (Sephadex G-75, superfine). 

Anion-exchange chromatography on a column of TSK DEAE-650 M (EM Science) in 30% methanol. Elution with a linear gradient of NaCl concentration from 0 to 1M. 

Step 4. Anion-exchange chromatography on a column of DEAE-Sephadex A-50. The column had been equilibrated with the basic buffer containing 0.12 M NaCl. The NaCl in the photoprotein solution was removed by gel filtration, and then the solution was added onto the column. The photoprotein adsorbed on the column was eluted with the equilibration buffer. 

The, chain voAiantS are characterized by the presence of two abnormal components, an abnormal Hb-F (02 /2) and an abnormal Hb-A (tt2 32) Of these two, the 02 2 component dominates and the 02 32 component Is often difficult to detect. The methods of choice are starch gel electrophoresis and anion-exchange chromatography using DEAE-Sephadex or DE-52 Cellulose. Chain analyses of these Isolated hemoglobin components will lead to a definitive Identification. 

Figure 3. Anion-exchange chromatography on DEAE Sepharose of fraction I of the Sephacryl S300 fractionation of a) saponified MHR population A after degradation with RGase and after b) degradation of the non-saponified MHR population A with RGase and RGAEase ---------, uronic acid —, neutral sugars thin line, NaOAc gradient [39],...

Table 4. Sugar composition (mol%) of the xylogalacturonan fractions obtained after DEAE Sepharose anion-exchange chromatography of the Sephacryl S300 fractions I originating from RGase degraded saponified apple MHR population A (1-a and 1-Z>) and from RGase/RGAEase treated non-saponified apple MHR population A (2-a, 2-b, and 2-c).

High performance anion-exchange chromatography evidence that the synthesized product is homogalacturonan... 

High-performance anion-exchange chromatography (HPAEC) of the reaction products from MHR-S showed that RGaseA and B acted differently towards this substrate (Fig. 1). 

Fig. 7. High-performance anion-exchange chromatography of the reaction products from alkali saponified SPS, produced by the crude A. acukatns preparation (Ultra-SP) and the partially purified fraction HTP-2.

Lee, YC (1990) High-performance anion-exchange chromatography for carbohydrate analysis. AnalBiochem 189 151-162... 

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