Abstract | BACKGROUND: OBJECTIVE: To characterize the biological functions of human anti-COL17 scFv antibody. METHODS: We constructed scFv antibodies against the corresponding antigen from parental Fab by expression in Escherichia coli. IgG autoantibodies against COL17 were purified by affinity chromatography from serum of BP patients. The inhibitory effects of anti-COL17 scFv on binding of BP autoantibodies to the NC16A domain of human COL17 antigen were observed by inhibition ELISA, immunofluorescence, and inhibition of complement activation. Reactive oxygen production assay and BP cryosection model were performed to assess the inhibitory effect of scFv on granulocyte activation and then the dermal-epidermal separation. RESULTS: ELISA and Western blot showed specific binding of scFv to COL17. We found that anti-COL17 scFv can inhibit the binding of intact IgG purified from BP parents to the corresponding COL17 antigen and then subsequent C1q and C3 activation and granulocyte activation in vitro. Most importantly, we confirmed that recombinant scFv can inhibit BP- IgG induced dermal-epidermal separation by BP cryosection model. CONCLUSION: The anti-COL17 scFv antibody can inhibit the binding of BP- IgG autoantibodies to COL17, thereby affecting subsequent complement activation and granulocyte activation in vitro. Our results suggest that blocking pathogenic epitopes using engineered scFv is an efficient BP therapy.
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Authors | Yan Wu, Na-Na Sun, Er-Le Dang, Liang Jin, Zhen-Feng Liu, Wei Zhang, Lu-Ting Yang, Gang Wang |
Journal | Journal of dermatological science
(J Dermatol Sci)
Vol. 72
Issue 1
Pg. 25-31
(Oct 2013)
ISSN: 1873-569X [Electronic] Netherlands |
PMID | 23827201
(Publication Type: Journal Article)
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Copyright | Copyright © 2013 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved. |
Chemical References |
- Antibodies, Blocking
- Autoantibodies
- Autoantigens
- Immunoglobulin G
- Non-Fibrillar Collagens
- Single-Chain Antibodies
- collagen type XVII
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Topics |
- Antibodies, Blocking
(administration & dosage, genetics)
- Antigen-Antibody Reactions
- Autoantibodies
(immunology)
- Autoantigens
(immunology)
- Autoimmunity
- Complement Activation
- Granulocytes
(immunology)
- Humans
- Immunoglobulin G
(immunology)
- Immunotherapy
- Non-Fibrillar Collagens
(antagonists & inhibitors, immunology)
- Pemphigoid, Bullous
(immunology, therapy)
- Protein Engineering
- Single-Chain Antibodies
(administration & dosage, genetics)
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