Abstract
A novel and efficient tagArray technology was developed that allows rapid identification of antibodies which bind to receptors with a specific expression profile, in the absence of biological information. This method is based on the cloning of a specific, short nucleotide sequence (tag) in the phagemid coding for each phage-displayed antibody fragment (phage-Ab) present in a library. In order to set up and validate the method we identified about 10,000 different phage-Abs binding to receptors expressed in their native form on the cell surface (10 k Membranome collection) and tagged each individual phage-Ab. The frequency of each phage-Ab in a given population can at this point be inferred by measuring the frequency of its associated tag sequence through standard DNA hybridization methods. Using tiny amounts of biological samples we identified phage-Abs binding to receptors preferentially expressed on primary tumor cells rather than on cells obtained from matched normal tissues. These antibodies inhibited cell proliferation in vitro and tumor development in vivo, thus representing therapeutic lead candidates.
Highlights
In recent years monoclonal antibodies have proven to be excellent therapeutic agents [1,2]
Since we aimed at identifying clones enriched in the T as compared to the matched N phage population, we focused our attention on tags exhibiting the highest T/N ratio in both patients
In the majority of cases the identification of a therapeutic target necessarily precedes the selection of a therapeutic monoclonal antibodies (mAbs), whereas we demonstrate that this ‘‘therapeutic target to mAb’’ approach can be reversed
Summary
In recent years monoclonal antibodies (mAbs) have proven to be excellent therapeutic agents [1,2] They have long half-life, favorable pharmaco-kinetics in humans, none or very few adverse reactions and a well established industry-scale production process [1,3,4]. We devised a highly sensitive method to survey the differential binding of a large number of clones which can be adapted to the very small scale of tissues biopsies This strategy allows the identification of epitopes with a specific expression profile (e.g., tumor-specific), independently of any biological information. The method is based on i) the availability of a defined collection of phage-Abs binding to the epitopes of membrane proteins (i.e. the Membranome collection) and ii) the possibility of tagging every phage-Ab with a specific DNA tag sequence
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