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ISSN
1672-9145
Acta Biochim Biophys Sin
2005, 37(6): 415–420
CN 31-1940/Q
Generation and Characterization of C305, a Murine Neutralizing scFv
Antibody That Can Inhibit BLyS Binding to Its Receptor BCMA
Mei-Yun LIU1,2, Wei HAN2,3, Yan-Li DING2,3,
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]