Combination of MS protein identification and bioassay of chromatographic fractions to identify biologically active substances from complex protein sources.

ABSTRACT: Purification of biologically active proteins from complex biological sources is a difficult task, usually requiring large amounts of sample and many separation steps. We found an active substance in a serum response element-dependent luciferase reporter gene bioassay in interstitial cystitis urine that we attempted to purify with column chromatography and the bioassay. With anion-exchange Mono Q and C4 reversed-phase columns, apparently sharp active peaks were obtained. However, more than 20 kinds of proteins were identified from the active fractions with MS, indicating that the purification was not complete. As further purification was difficult, we chose a candidate molecule by means of studying the correlation between MS protein identification scores and bioassay responses of chromatogr

SUBMITTER: Kuromitsu S 

PROVIDER: S-EPMC2690494 | biostudies-literature | 2009 Jun

REPOSITORIES: biostudies-literature