Abstract
A system consisting of high-performance affinity chromatography and size-exclusion chromatography has been developed for the rapid purification and isolation of relatively labile membrane proteins, such as growth-hormone receptor. The crude membrane sample containing growth-hormone receptor was obtained from rabbit livers by ultracentrifugation, followed by solubilization with Triton X-100. The sample was introduced into the high-performance affinity gel column through a large-volume loop injector. After removal of unretained proteins, the fraction containing the growth-hormone receptor was eluted with 6 M urea solution. The eluate from the affinity column passed directly into the size-exclusion gel column, where rapid desalting and separation from contaminants were achieved. The eluate was monitored by UV absorption at 280 nm and fractionated for the subsequent binding assay with 125I-labelled human growth hormone.
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