Detection of Elution Profile of Protein in the Presence of Phenyl-Based Detergent during High-Performance Liquid Chromatography of Cytochrome P-450(Biological,Chemical)

概要

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Cytochrome P-450 (P-450) isozyrnes are well separated by high-performance liquid chromatography with an ion-exchange column using buffer containing 0.4% Emulgen 911 (polyphenyl ether) as a detergent. However, it is impossible to monitor the elution profile of protein at 280 nm in the presence of Emulgen. Thus, we have developed a technique for determining the elution profile of protein in the presence of Emulgen. The absorption maximum and minimum of Emulgen 911 were at 276 and 244 nm, respectively. At 244 nm, the elution profile of 0.5 nmol (35μg) of P-450 was detected as a peak in the presence of 0.4% Emulgen 911. The peak area increased linearly with increasing amount of P-450 up to 5 nmol (350μg). Identification and estimation of the purity of the eluted P-450 are possible by dual monitoring at 244 and 417 nm, the wavelength for the detection of hemoprotein.
社団法人日本薬学会の論文
1987-12-25