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Anal Biochem ; 406(2): 147-56, 2010 Nov 15.
Article in English | MEDLINE | ID: mdl-20624370

ABSTRACT

Therapeutic monoclonal antibodies are becoming a significant and rapidly growing class of therapeutic pharmaceuticals. Their discovery and development requires fast and high-throughput methodologies for screening and selecting appropriate candidate antibodies having high affinity for the target as well as high specificity and low cross-reactivity. This study demonstrates the use of the ProteOn XPR36 protein interaction array system and its novel approach, termed One-Shot Kinetics, for the rapid screening and selection of high-affinity antibodies. This approach allows multiple quantitative protein binding analyses in parallel, providing association, dissociation, and affinity constants for several antibodies or supernatants simultaneously in one experiment. We show that the ProteOn XPR36 system is a valuable tool for use across multiple stages of the therapeutic antibody discovery and development process, enabling efficient and rapid screening after panning, affinity maturation, assay validation, and clone selection.


Subject(s)
Antibodies, Monoclonal/therapeutic use , Biosensing Techniques/instrumentation , Biosensing Techniques/methods , Animals , Antibodies, Monoclonal/immunology , Antibody Affinity/immunology , Antigens/immunology , CHO Cells , Clone Cells , Cricetinae , Cricetulus , Humans , Immunoglobulin Fab Fragments/immunology , Immunoglobulin G/immunology , Kinetics , Mutation/genetics , Protein Binding , Receptors, IgG/immunology , Reproducibility of Results
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