[Expression, purification and activity analysis of human single-chain antibody against hepatocellular cancer]

[Expression, purification and activity analysis of human single-chain antibody against hepatocellular cancer] Research Abstract Details 

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[Expression, purification and activity analysis of human single-chain antibody against hepatocellular cancer] Abstract Text:

Qing-tao Zhao,Long-yang Ma,Guo-zhu Xue,Ai-zhi Zhao,Ke-feng Dou,

AIM: To express and characterize an active form of a single-chain antibody (scFv) from the gene of human phage antibodies which is specific for hepatocellular cancer. METHODS: The complementary DNAs encoding the variable regions of the light chain (V(L)) and heavy chain (V(H)) were connected by a (Gly(4)Ser)(3) linker using a splicing by overlap extension polymerase chain reaction. The resultant scFv gene was cloned to the pET28a(+) vector and expressed in E.coli as inclusion bodies. Then the inclusion bodies were solubilized, denatured and renatured. Finally, the affinity constant of scFv was determined by noncompetitive enzyme immunoassay. RESULTS: The target protein amounted 26% of the total protein in the condition of A(600) at 0.8 for 6 hours. After renatured, the purity of target protein was 95% and the affinity constant of the scFv was 3.6x10(7) mol/L. CONCLUSION: An active form of scFv which is specific for hepatocellular cancer can bind selectively with hepatocellular cancer cells, which provides a theoretical basis for immunological detection and clinical use of scFv.

[Expression, purification and activity analysis of human single-chain antibody against hepatocellular cancer] Publishing Authors By Initials

QT Zhao,LY Ma,GZ Xue,AZ Zhao,KF Dou,

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[Expression, purification and activity analysis of human single-chain antibody against hepatocellular cancer] Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese jour

VOLUME: 23

Page Numbers: 649-51

Journal Abbreviation: Xi Bao Yu Fen Zi Mian Yi Xue Z

ISSN: 1007-8738

DAY: 9

MONTH: Jul

YEAR: 2007

[Expression, purification and activity analysis of human single-chain antibody against hepatocellular cancer] Information

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LANGUAGE: chi

NlmUniqueID: 101139110

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AFFILIATION: Department of Hepatobiliary Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

Country: China

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MEDLINETA: Xi Bao Yu Fen Zi Mian Yi Xue Z

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