Document Detail


Effect of domain order on the activity of bacterially produced bispecific single-chain Fv antibodies.
MedLine Citation:
PMID:  12818205     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Bispecific single-chain Fv antibodies comprise four covalently linked immunoglobulin variable (VH and VL) domains of two different specificities. Depending on the order of the VH and VL domains and on the length of peptides separating them, the single-chain molecule either forms two single-chain Fv (scFv) modules from the adjacent domains of the same specificity, a so-called scFv-scFv tandem [(scFv)(2)], or folds head-to-tail with the formation of a diabody-like structure, a so-called bispecific single-chain diabody (scBsDb). We generated a number of four-domain constructs composed of the same VH and VL domains specific either for human CD19 or CD3, but arranged in different orders. When expressed in bacteria, all (scFv)(2) variants appeared to be only half-functional, binding to CD19 and demonstrating no CD3-binding activity. Only the diabody-like scBsDb could bind both antigens. Comparison of the scBsDb with a structurally similar non-covalent dimer (diabody) demonstrated a stabilizing effect of the linker in the middle of the scBsDb molecule. We demonstrated that the mechanism of inactivation of CD19xCD3 diabody under physiological conditions is initiated by a dissociation of the weaker (anti-CD3) VH/VL interface followed by domain swapping with the formation of non-active homodimers. The instability of one homodimer makes the process of diabody dissociation/reassociation irreversible, thus gradually decreasing the fraction of active molecules. The structural parameters influencing the formation of functional bispecific single-chain antibodies are indicated and ways of making relatively stable bispecific molecules are proposed.
Authors:
Sergey M Kipriyanov; Gerhard Moldenhauer; Michael Braunagel; Uwe Reusch; Björn Cochlovius; Fabrice Le Gall; Olga A Kouprianova; Claus Wilhelm Von der Lieth; Melvyn Little
Related Documents :
3007145 - Human complement component c1s. partial sequence determination of the heavy chain and i...
6427235 - Mouse myeloma cells that make short immunoglobulin heavy chains: pleiotropic effects on...
2422275 - Novel human light chain v kappa segment: serologic and structural analyses of the kappa...
1709665 - Junctional diversity of h and l chains allows the coexpression of two mutually exclusiv...
181085 - A kinetic analysis of the dephosphorylation, by bovine spleen phosphoprotein phosphatas...
7627335 - Calcitonin, calcitonin gene-related peptide, adrenomedullin and amylin: homologous pept...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of molecular biology     Volume:  330     ISSN:  0022-2836     ISO Abbreviation:  J. Mol. Biol.     Publication Date:  2003 Jun 
Date Detail:
Created Date:  2003-06-23     Completed Date:  2003-07-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985088R     Medline TA:  J Mol Biol     Country:  England    
Other Details:
Languages:  eng     Pagination:  99-111     Citation Subset:  IM    
Affiliation:
Recombinant Antibody Research Group, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany. s.kipriyanov@affimed.com
Data Bank Information
Bank Name/Acc. No.:
PDB/1OSO;  1OSQ
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Antibodies, Bispecific / chemistry*,  genetics,  metabolism*,  pharmacology
Antigens / metabolism
Antigens, CD19 / genetics
Antigens, CD3 / genetics
Dimerization
Drug Screening Assays, Antitumor
Drug Stability
Escherichia coli / genetics
Humans
Immune Sera
Immunoglobulin Fragments / chemistry*,  genetics,  metabolism*,  pharmacology
Immunoglobulin Variable Region / chemistry,  metabolism,  pharmacology
Molecular Sequence Data
Protein Engineering / methods
Protein Structure, Tertiary / genetics,  physiology
Structure-Activity Relationship
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Antibodies, Bispecific; 0/Antigens; 0/Antigens, CD19; 0/Antigens, CD3; 0/Immune Sera; 0/Immunoglobulin Fragments; 0/Immunoglobulin Variable Region; 0/immunoglobulin Fv

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


Previous Document:  Structure of the large subunit of class Ib ribonucleotide reductase from Salmonella typhimurium and ...
Next Document:  The active conformation of glutamate synthase and its binding to ferredoxin.