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Internalizing Antibody Screening

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Abstract or Description:

Internalizing antibodies and peptides specific for cell surface receptors, which upon binding will induce receptor-mediated endocytosis, are highly suitable vehicles for targeted delivery of drugs, toxins, enzymes or DNA into the cytosol of mammalian cells for therapeutic applications. Compared with receptor-independent antibodies and peptides that can penetrate cell membrane without binding to a receptor, receptor-mediated internalizing antibodies and peptides confer cell-specific drug delivery, the fundamental requirement for targeted therapy. Selection of receptor-mediated internalizing antibodies and peptides from phage display library can also be employed to identify cell-specific markers, the endocytosed receptors that are associated with a specific cellular function.
 
We have done a large number of whole-cell based phage display library screening projects to select internalizing antibodies and peptides specific for various cell surface receptors such as EGFR, NTRK2, CTLA4, HER2, CD20, CD3, CD59, transferrin receptor and CD33. Our staff scientists also have extensive experience in discovery of novel marker receptors by selecting internalizing antibodies and peptides that are specific for certain cell types. Methodologies are developed to reduce the number of internalizing phage particles that are not target receptor-specific, to exclude phage particles that are merely bound to the cell surface and to selectively identify phage particles that are capable of crosslinking receptors, rather than merely binding. 
 
We found that receptor-dependent internalizing antibodies and peptides are sometimes cargo-dependent as well. An internalizing antibody/peptide that permits internalization of one cargo may not support the internalization of a different cargo. Therefore, in order to select the best internalizing antibodies or peptides for a defined cargo, a custom cargo-fusion antibody/peptide library is necessary. We employ both filamentous and T7 lambda phage systems to make cargo-fusion antibody and peptide libraries in selection of internalizing antibodies and peptides, especially for toxins, enzymes and other protein cargos. 

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Posted By: Candy Ton on 6/9/2015 8:41:04 AM
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