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Efficient high-performance liquid chromatographic system for protein purification.

S Yagi·K Izawa·T Nakagawa·H Tanaka·A Yoshitake·Z Mohri
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Abstract

An efficient high-performance liquid chromatographic system, consisting of an affinity column and a high-performance size-exclusion column, was developed and applied to the purification of growth hormone receptors from rabbit livers. When a 6-ml sample of Triton X-100 extracts containing 16 mg of protein was applied to the system, 1200-fold purified receptor with a 10% recovery of binding activity from homogenates was obtained within 3-4 h. The purified receptor exhibited one main band on sodium dodecyl sulphate polyacrylamide gel electrophoresis, and the affinity constant (Ka = 6.0.10(9) M-1) was found to be comparable with that of 1% Triton X-100 extract (4.4.10(9) M-1). The injection of 1 ml of 3 M urea solution prior to receptor elution with 10 ml of 6 M urea solution was effective in removing non-specific binding proteins.

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AnimalsChromatography, AffinityChromatography, High Pressure LiquidElectrophoresis, Polyacrylamide GelLiverProteinsRabbitsReceptors, SomatotropinSpectrophotometry, UltravioletJournal ArticleResearch Support, Non-U.S. Gov't

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S Yagi, K Izawa, T Nakagawa, H Tanaka, A Yoshitake, Z Mohri. 1989-08-25. Efficient high-performance liquid chromatographic system for protein purification.. https://doi.org/10.1016/s0378-4347(00)82705-x

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