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J Immunol Methods ; 354(1-2): 85-90, 2010 Mar 31.
Article in English | MEDLINE | ID: mdl-20153332

ABSTRACT

Recombinant monoclonal antibodies currently dominate the protein biologics marketplace. The path from target antigen discovery and screening, to a recombinant therapeutic antibody can be time-consuming and laborious. We describe a set of expression vectors, termed mAbXpress, that enable rapid and sequence-independent insertion of antibody variable regions into human constant region backbones. This method takes advantage of the In Fusion cloning system from Clontech, which allows ligation-free, high-efficiency insertion of the variable region cassette without the addition of extraneous amino acids. These modular vectors simplify the antibody reformatting process during the preliminary evaluation of therapeutic or diagnostic candidates. The resulting constructs can be used directly for transient or amplifiable, stable expression in mammalian cells. The effectiveness of this method was demonstrated by the creation of a functional, fully human anti-human CD83 monoclonal antibody.


Subject(s)
Antibodies, Monoclonal/immunology , Antigens, CD/immunology , Cloning, Molecular , Genetic Vectors , Immunoglobulin Constant Regions/immunology , Immunoglobulin Variable Region/immunology , Immunoglobulins/immunology , Membrane Glycoproteins/immunology , Peptide Library , Animals , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/genetics , Antibody-Dependent Cell Cytotoxicity , Binding Sites, Antibody , CHO Cells , Cricetinae , Cricetulus , Flow Cytometry , Humans , Immunoglobulin Constant Regions/biosynthesis , Immunoglobulin Constant Regions/genetics , Immunoglobulin Variable Region/biosynthesis , Immunoglobulin Variable Region/genetics , Killer Cells, Lymphokine-Activated/immunology , Recombinant Proteins/immunology , Time Factors , Transfection , CD83 Antigen
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