Tetanus Neurotoxin Neutralizing Antibodies Screened from a Human Immune scFv Antibody Phage Display Library.
Tetanus neurotoxin (TeNT) produced by Clostridium tetani is one of the most poisonous protein substances. Neutralizing antibodies against TeNT can effectively prevent and cure toxicosis. Using purified Hc fragments of TeNT (TeNT-Hc) as an antigen, three specific neutralizing antibody clones recognizing different epitopes were selected from a human immune scFv antibody phage display library. The three antibodies (2-7G, 2-2D, and S-4-7H) can effectively inhibit the binding between TeNT-Hc and differentiated PC-12 cells in vitro. Moreover, 2-7G inhibited TeNT-Hc binding to the receptor via carbohydrate-binding sites of the W pocket while 2-2D and S-4-7H inhibited binding of the R pocket. Although no single mAb completely protected mice from the toxin, they could both prolong survival when challenged with 20 LD50s (50% of the lethal dose) of TeNT. When used together, the mAbs completely neutralized 1000 LD50s/mg Ab, indicating their high neutralizing potency in vivo. Antibodies recognizing different carbohydrate-binding pockets could have higher synergistic toxin neutralization activities than those that recognize the same pockets. These results could lead to further production of neutralizing antibody drugs against TeNT and indicate that using TeNT-Hc as an antigen for screening human antibodies for TeNT intoxication therapy from human immune antibody library was convenient and effective.
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. happywanghan@163.com.|Clinical Diagnostic Centre, 302 Military Hospital of China, Beijing 100039, China. happywanghan@163.com.
Han
H
Wang
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. yurui1102@139.com.
Rui
R
Yu
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. 15510742074@163.com.
Ting
T
Fang
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. wanghan129@sina.com.
Ting
T
Yu
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. goodnightcxy@163.com.
Xiangyang
X
Chi
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. xienawh@sina.com.
Xiaopeng
X
Zhang
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. 15210215727@163.com.
Shuling
S
Liu
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. 15011221815@163.com.
Ling
L
Fu
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. yuchangming@126.com.
Changming
C
Yu
Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, Beijing 100071, China. CW0226@foxmail.com.
Wei
W
Chen
D000818:Animals; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D000915:Antibody Affinity; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D061505:Cell Surface Display Techniques; D004195:Disease Models, Animal; D000939:Epitopes; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000276:immunology / Q000378:metabolism; D016716:PC12 Cells; D010446:Peptide Fragments / Q000276:immunology* / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D051381:Rats; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D013742:Tetanus / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D013744:Tetanus Toxin / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors
D057134:Antibodies, Neutralizing; D000939:Epitopes; D008565:Membrane Proteins; D010446:Peptide Fragments; D019151:Peptide Library; D057127:Single-Chain Antibodies; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C; C028728:tetanus toxin receptor
Hc fragment; human immune scFv antibody phage display library; neutralizing antibody; phage display; tetanus neurotoxins