Publication: Development of an oxalate-affinity chromatographic column to isolate oxalate-binding proteins
Issued Date
2010-08-01
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ISSN
17599679
17599660
17599660
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2-s2.0-77958099274
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Mahidol University
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SCOPUS
Bibliographic Citation
Analytical Methods. Vol.2, No.8 (2010), 1051-1055
Suggested Citation
Piyachat Roop-Ngam, Visith Thongboonkerd Development of an oxalate-affinity chromatographic column to isolate oxalate-binding proteins. Analytical Methods. Vol.2, No.8 (2010), 1051-1055. doi:10.1039/b9ay00285e Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/28893
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Title
Development of an oxalate-affinity chromatographic column to isolate oxalate-binding proteins
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Abstract
Calcium oxalate (CaOx) is the major crystalline component in kidney stones. Many previous studies on stone modulation have focused mainly to calcium-binding proteins, whereas oxalate-binding proteins remain under-investigated due to a lack of a simple method to purify oxalate-binding proteins. Therefore, we have developed an affinity chromatographic column to isolate or purify oxalate-binding proteins. The oxalate-affinity column developed contained EAH-Sepharose 4B beads conjugated with oxalic acid by carbodiimide reaction using 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) as an activator. The coupling efficacy of coupled beads (oxalic acid-EAH Sepharose 4B) was determined by a competitive ninhydrin assay to quantify the remaining amino group on EAH-Sepharose 4B. In addition, the oxalate-affinity column showed high specific binding to a known oxalate-binding protein (p62) but not to an irrelevant protein (carbonic anhydrase). In conclusion, we have established the oxalate-affinity chromatographic column, which showed the high efficiency to isolate oxalate-binding proteins. This novel chromatographic column will be very useful for future stone research, particularly in the area of stone modulation by oxalate-binding proteins. © 2010 The Royal Society of Chemistry.