Document Detail


Covalent dimerization of camelidae anti-human TNF-alpha single domain antibodies by the constant kappa light chain domain improves neutralizing activity.
MedLine Citation:
PMID:  20047190     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The tumor necrosis factor-alpha (TNFalpha) plays an important role in a number of chronic inflammatory disorders. Monoclonal camelidae variable heavy chain domain-only antibodies (V(H)H) have been developed to antagonize the action of human TNFalpha (anti-TNF-V(H)H). Here we describe a strategy to obtain functional dimeric anti-TNF-V(H)H molecules, based on the C-terminal fusion of a kappa light chain domain to the anti-TNF-V(H)H. The resulting fusion protein was transiently expressed by use of viral vectors in Nicotiana benthamiana((Nb)) leaves and purified. Competitive ELISA and cell cytotoxicity assays revealed that the dimerized anti-(Nb)TNF-V(H)H(Ckappa) proteins blocked TNFalpha-activity more effectively than either the monomeric Escherichia coli((Ec)) produced anti-(Ec)TNF-V(H)H or the monomeric anti-(Nb)TNF-V(H)H(Ckappa). We suggest that enhanced inhibition shown by dimeric anti-(Nb)TNF-V(H)H(Ckappa) proteins is achieved by an increase in avidity.
Authors:
Martin Giersberg; Doreen M Floss; Sergey Kipriyanov; Udo Conrad; J?rgen Scheller
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  106     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-03-29     Completed Date:  2010-06-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  161-6     Citation Subset:  IM    
Affiliation:
Institute of Plant Genetics and Crop Plant Research (IPK), Phytoantibodies, D-06466 Gatersleben, Germany.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal / genetics,  pharmacology*
Antibodies, Neutralizing / genetics,  pharmacology*
Dimerization
Gene Expression
Genetic Vectors
Humans
Immunoglobulin Light Chains / genetics,  pharmacology*
Immunologic Factors / genetics,  pharmacology*
Plant Viruses / genetics
Recombinant Fusion Proteins / genetics,  pharmacology
Tobacco / genetics
Tumor Necrosis Factor-alpha / antagonists & inhibitors*
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antibodies, Neutralizing; 0/Immunoglobulin Light Chains; 0/Immunologic Factors; 0/Recombinant Fusion Proteins; 0/Tumor Necrosis Factor-alpha

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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