Generation of Three Selenium-Containing Catalytic Antibodies with High Catalytic Efficiency Using a Novel Hapten Design Method

2001 ◽  
Vol 395 (2) ◽  
pp. 177-184 ◽  
Author(s):  
Dan Su ◽  
Xiaojun Ren ◽  
Delin You ◽  
Dan Li ◽  
Ying Mu ◽  
...  
Keyword(s):  
Design Method ◽  
Catalytic Efficiency ◽  
Catalytic Antibodies ◽  
Hapten Design

Catalytic Antibodies for Complex Reactions Hapten Design and the Importance of Screening for Catalysis in the Generation of Catalytic Antibodies for the NDA/CN Reaction

2000 ◽  
Vol 83 (1-3) ◽  
pp. 195-208 ◽  
Author(s):  
Binodh S. Desilva ◽  
Gyorgy Orosz ◽  
Kamal L. Egodage ◽  
Robert G. Carlson ◽  
Richard L. Schowen ◽  
...  
Keyword(s):  
Catalytic Antibodies ◽  
Hapten Design

Catalytic antibodies: hapten design strategies and screening methods

2004 ◽  
Vol 12 (20) ◽  
pp. 5247-5268 ◽  
Author(s):  
Yang Xu ◽  
Noboru Yamamoto ◽  
Kim D. Janda
Keyword(s):  
Catalytic Antibodies ◽  
Screening Methods ◽  
Design Strategies ◽  
Hapten Design

scholarly journals Augmenting the efficacy of anti-cocaine catalytic antibodies through chimeric hapten design and combinatorial vaccination

2017 ◽  
Vol 27 (16) ◽  
pp. 3666-3668 ◽  
Author(s):  
Cody J. Wenthur ◽  
Xiaoqing Cai ◽  
Beverly A. Ellis ◽  
Kim D. Janda
Keyword(s):  
Catalytic Antibodies ◽  
Hapten Design

Cloning and expression of a single-chain catalytic antibody that acts as a glutathione peroxidase mimic with high catalytic efficiency

2001 ◽  
Vol 359 (2) ◽  
pp. 369-374 ◽  
Author(s):  
Xiaojun REN ◽  
Shujuan GAO ◽  
Delin YOU ◽  
Hualiang HUANG ◽  
Zi LIU ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Glutathione Peroxidase ◽  
Binding Site ◽  
Active Form ◽  
Catalytic Efficiency ◽  
Catalytic Antibodies ◽  
Rabbit Liver ◽  
Cloning And Expression ◽  
Single Chain ◽  
Variable Regions

Glutathione peroxidase (GPX) has a powerful role in scavenging reactive oxygen species. In previous papers we have developed a new strategy for generating abzymes: the monoclonal antibody with a substrate-binding site is first prepared, then a catalytic group is incorporated into the monoclonal antibody's binding site by using chemical mutation [Luo, Zhu, Ding, Gao, Sun, Liu, Yang and Shen (1994) Biochem. Biophys. Res. Commun. 198, 1240–1247; Ding, Liu, Zhu, Luo, Zhao and Ni (1998) Biochem. J. 332, 251–255]. Since then we have established a series of catalytic antibodies capable of catalysing the decomposition of hydroperoxides by GSH. The monoclonal antibody 2F3 was raised against GSH-S-2,4-dinitrophenyl t-butyl ester and exhibited high catalytic efficiency, exceeding that of rabbit liver GPX, after chemical mutation. To produce pharmaceutical proteins and to study the reason why it exhibits high catalytic efficiency, we sequenced, cloned and expressed the variable regions of 2F3 antibody as a single-chain Fv fragment (2F3-scFv) in different bacterial strains. The amounts of 2F3-scFv proteins expressed from JM109 (DE3), BL21 (DE3), and BL21 (coden plus) were 5–10%, 15–20% and 25–30% of total bacterial proteins respectively. The 2F3-scFv was expressed as inclusion bodies, purified in the presence of 8M urea by Co2+-immobilized metal-affinity chromatography (IMAC) and renatured to the active form in vitro by gel filtration. The binding constants of the active 2F3-scFv for GSH and GSSG were 2.46×105M−1 and 1.03×105M−1 respectively, which were less by one order of magnitude than that of the intact 2F3 antibody. The active 2F3-scFv was converted into selenium-containing 2F3-scFv (Se-2F3-scFv) by chemical modification of the reactive serine; the GPX activity of the Se-2F3-scFv was 3394units/μmol, which approaches the activity of rabbit liver GPX.

Catalytic Antibodies: Hapten Design Strategies and Screening Methods

ChemInform ◽  
2005 ◽  
Vol 36 (6) ◽  
Author(s):  
Yang Xu ◽  
Noboru Yamamoto ◽  
Kim D. Janda
Keyword(s):  
Catalytic Antibodies ◽  
Screening Methods ◽  
Design Strategies ◽  
Hapten Design

Application of a work-centered design method to support counterspace operations

2005 ◽  
Author(s):  
Michael Szczepkowski ◽  
Kelly Neville ◽  
Ed Popp
Keyword(s):  
Design Method
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