• FOCUS: LIFE SCIENCE
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• no.1
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• pp.5.1-5.3
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• 2024

The document discusses the regeneration of Partisil/Partisphere ion-exchange columns in chromatography. It mentions that column efficiency can diminish with use due to the accumulation of sample and/or mobile phase impurities at the head of the column. This can lead to a change in back pressure, lower column efficiency, and sometimes a change in selectivity. The document outlines a procedure that may restore column performance. The document also provides everyday practices to enhance the lifetime of a column. These include using only high-purity HPLC solvents and buffers, using freshly prepared mobile phases and buffers, filtering mobile phases to remove particulates, using appropriate sample clean-up procedures, using a guard column or pre-column filter, and working within the pressure and flow rate limitations of the column. For the regeneration of Partisil/Partisphere SAX, SCX, WAX, and WCX columns, the document suggests passing 20 column volumes of various mobile phases through the column. These include a buffer wash, distilled water, an acid wash, a chelating wash, a methanol wash, and a buffer for separation. The document emphasizes that not all of these wash steps are required for every column clean-up and that some chromatographers require only a combination of certain steps.

• Korean Chemical Engineering Research
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• v.50 no.4
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• pp.659-665
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• 2012

IgY (Immunoglobulin Yolk) in egg yolk corresponds to IgG (Immunoglobulin G) in animal serum and plays an important role as immunological proteins in intestines. Carrageenan and Arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE (Diethylaminoethyl) Sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtration High Performance Liquid Chromatography) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatography) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at 0.5 M NaCl and pH=5.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.343-347
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• 2005

The polysaccharides were extracted from fruiting body, mycelia, and cell-free broth of Agaricus blazei Murill. The crude polysaccharides were obtained by the ethanol addtion. They were further purified using ion-exchange chromatography and gel chromatography. Ion-exchange chromatography using DEAE-cellulose column separated neutral and acidic polysaccharides. Neutral polysaccharides were then purified with gel filtration chromatography. For single peak obtained from gel filtration chromatography was molecular weight was measured with Sepharose CL-6B. The same procedure with acidic polysaccharides were performed to get the purified polysaccharides.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.254-259
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• 2002

A psychrotrophic bacterium was isolated from Antarctic marine sediment and identified as Shewanella sp. species based on the biochemical properties and 16S rRNA sequence, and designated as Shewanella sp. L93. Extracellular protease produced by this strain was purified through ammonium sulfate precipitation, High-Q column chromatography, first gel permeation chromatography, BioScale Q2 ion exchange chromatography and second gel permeation chromatography, and basic properties of this enzyme were investigated.

• Korean Journal of Food Science and Technology
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• v.24 no.6
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• pp.616-618
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• 1992

The study was attempted to isolate taurine from the oyster shucking juice known as one of the by-products of oyster processing using ion exchange column chromatography. Three hundred grams of the oyster shucking juice were loaded onto a column packed with 300 ml of the Dowex 50W $H^+$ form. And taurine-rich fractions were further purified in columns packed with 150 ml of Dowex 2 OH form and the 150 ml of Amberite IRA-410 OH form consecutively. The purity and the yield of taurine recovered from the oyster shucking juice by this method were 94.7% and 84.8%, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.117-121
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• 1997

Glutamic acid can be crystallized inside cation exchange column when displacer NaOH concentration is high enough to concentrate displaced glutamic acid beyond its solubility limit. Resulting crystal layer of glutamic acid was moved with liquid phase through the column, and thus could be eluted from the column and recovered in fraction collector. For the purpose of enhancing crystal recovery, effects of operating parameters on the crystal formation were investigated. The increase in the degree of crosslinking of resin favored crystal recovery because of its low degree of swelling. Higher concentration of displacer NaOH was advantageous. If NaOH concentration is too high, however, crystal recovery was lowered due to the solubility-enhancing effects of high pH and ionic strength. The decrease of mobile phase flow rate enhanced crystal recovery because enough time to attain local equilibrium could be provided, but film diffusion would control the overall crystal formation with extremely low flow rate. Lower temperature reduced solubility of glutamic acid and thus favored crystal formation unless the rate of ion exchange was severely reduced. The ion exchange operated by displacement mode coupled with crystallization was advantageous in reducing the burden of further purification steps and in preventing purity-loss resulted from overlapping between adjacent bands.

• Bulletin of the Korean Chemical Society
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• v.24 no.9
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• pp.1324-1328
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• 2003

The simultaneous determination of anions ($SO_4 ^{2-},\;Cl^-,\;and\;NO_3^-$) and cations ($Na^+,\;NH^{4+},\;K^+,\;Mg^{2+},\;and\;Ca^{2+}$) in natural water obtained by Nakdong River waters system in Korea were performed by ion-exclusion/cationexchange chromatography with conductimetric detection. The stationary phase was a polymethacrylate-based weakly acidic cation-exchange resin column in the $H^+$-form and a weak-acid eluent. When using only a 1.4 mM sulfosalicylic acid/6 mM 18-crown-6 ether as an eluent, good resolution of both anions and cations, minimum time required for the separation, and satisfactory detection sensitivity were obtained in a reasonable time. The method was successfully applied to the simultaneous determination of anions and cations in natural waters.

• Microbiology and Biotechnology Letters
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• v.23 no.4
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• pp.485-491
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• 1995

Protein ion exchange chromatography was studied experimentally in order to prove the theoretical prediction from the linear model of Yamamoto, S. et al (1). Adsorption isotherms were measured as a function of ionic strength in a batch experiment. The relationship between the characteristics of chromatogram and the operating conditions of ionic strength, flow rate, length of column, concentration and amount of protein sample were studied. At the higher ionic strength, the lower flow rate and the longer column conditions, the higher number of plate was obtained. Satisfactory agreement was observed between the experimental and the calculated chromatograms except for the case of high protein concentration.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.45-45
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• 1993

괄루근으로부터 분리된 다당류에 대하여 continuous gel electrophoresis, SDS-Polyacrylamide gel eletrophoresis, ion exchange column chromatography, Hydroxyapatite column chromatography 및 Gel filtration등의 방법을 이용하여 다음과 같은 결과를 얻었다. 1) 황산암모늄 분별침전법에 의한 렉틴의 정제도는 초추출물의 4.85배이며 DEAE Sephadex A-50 column chromatography법에 의한 정제도는 24.17배로 나타났고, 마지막 정제단계인 Sephacryl S-200 gel filtration에 의한 정제도는 47.34배로 나타났다. 2) 정제된 렉틴의 분자량은 60,000da1ton으로 나타났다. 3) 사람의 혈액형에 따른 응집효과는 90-100％로 특이성은 없었다.

• Bulletin of the Korean Chemical Society
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• v.18 no.6
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• pp.648-653
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• 1997

Acetolactate synthase was purified from Escherichia coli MF2000/pTATX containing Arabidopsis thaliana acetolactate synthase gene. Purification steps included DEAE cellulose ion exchange column chromatography, phenyl sepharose hydrophobic column chromatography, hydroxylapatite affinity column chromatography, and Mono-Q HPLC. Molecular weight was estimated to be ∼65 KDa and purification fold was 109 times. The enzyme showed a pH optimum of 7 and the $K_M$ value was 5.9 mM. The purified enzyme was not inhibited by any of the end products, valine, leucine, and isoleucine.