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ISSN 1672-9145 Acta Biochim Biophys Sin 2005, 37(6): 415–420 CN 31-1940/Q

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Generation and Characterization of C305, a Murine Neutralizing scFv Antibody That Can Inhibit BLyS Binding to Its Receptor BCMA

Mei-Yun LIU^1,2, Wei HAN^2,3, Yan-Li DING^2,3, Tian-Hong ZHOU¹, Rui-Yang TIAN², Sheng-Li YANG², Hui LIU²*, and Yi GONG²*

¹ College of Life Science and Technology, Jinan University, Guangzhou 510632, China;

² Shanghai Research Center of Biotechnology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200233, China;

³ School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China

Abstract B-lymphocyte stimulator (BLyS) is a member of the tumor necrosis factor (TNF) family and a key regulator of B cell response. Neutralizing single-chain fragment variable (scFv) antibody against BLyS binding to its receptor BCMA has the potential to play a prominent role in autoimmune disease therapy. A phage display scFv library constructed on pIII protein of M13 filamentous phage was screened using BLyS. After five rounds of panning, their binding activity was characterized by phage-ELISA. Nucleotide sequencing revealed that at least two different scFv gene fragments (C305 and D416) were obtained. The two different scFv gene fragments were expressed to obtain the soluble scFv antibodies, then the soluble scFv antibodies were characterized by means of competitive ELISA and in vitro neutralization assay. The results indicated that C305 is the neutralizing scFv antibody that can inhibit BLyS binding to its receptor BCMA.

Key words phage display; neutralizing; scFv antibody; BLyS

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Received: February 24, 2005 Accepted: April 14, 2005

*Corresponding authors:

Yi GONG: Tel, 86-21-64369607; Fax, 86-21-64700244; E-mail, [email protected]

Hui LIU: Tel, 86-21-54971071; Fax, 86-21-64700244; E-mail, [email protected]