Purification and activity of human fetal hepatic stimulating substance

doi:10.3748/wjg.v2.i4.194

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

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World J Gastroenterol. Dec 15, 1996; 2(4): 194-196
Published online Dec 15, 1996. doi: 10.3748/wjg.v2.i4.194

Purification and activity of human fetal hepatic stimulating substance

Wei-Jian Hou, Su-Yuan Liu, Wei Xing, Zhong-Cheng Pan, Jin-Lan Liu, Dian-Hong Wang

Wei-Jian Hou, Su-Yuan Liu, Wei Xing, Zhong-Cheng Pan, Dian-Hong Wang, Department of Biochemistry, School of Basic Medical Sciences, China Medical University, Shenyang 110001, Liaoning Province, China

Jin-Lan Liu, Department of Gastroenterology, First Clinical College, China Medical University, Shenyang 110001, Liaoning Province, China

Author contributions: All authors contributed equally to the work.

Supported by The National Natural Science Foundation of China, No. 38970403.

Correspondence to: Dr. Dian-Hong Wang, Professor, Department of Biochemistry, School of Basic Medical Sciences, China Medical University, Shenyang 110001, Liaoning Province, China

Received: July 26, 1996
Revised: September 15, 1996
Accepted: November 10, 1996
Published online: December 15, 1996

Abstract

AIM: To purify human fetal hepatic stimulating substance (hHSS) and to determine its activity.

METHODS: hHSS was purified by ion exchange chromatography, nondenaturing polyacrylamide gel electrophoresis (PAGE), and ISCO (In situ chemical oxidation) concentrator electrophoresis. The molecular weight was determined by sodium dodecyl sulfate (SDS)-PAGE and the activity of hHSS was examined by both ³H-thymidine incorporation assay and enzyme linked immunosorbent assay (ELISA).

RESULTS: The purity of hHSS was increased along with the purification process. After many steps, the major band of nondenaturing PAGE showed a high activity and purity, and two bands were revealed by SDS-PAGE.

CONCLUSION: The molecular weight of hHSS is about 70 kDa, and it consists of two subunits with molecular weights of 18 kDa and 53 kDa, respectively. The activity of hHSS was assayed by both ³H-TdR incorporation assay and ELISA, and good consistence was seen.

Keywords: Liver stimulating; Substance; Molecular weight