We have used a naive human single-chain fragment variable (scFv) library as a source of random shape repertoire to directly
probe the altered surface chemistry of tumor cells. We reported previously the identification of more than 90 internalizing
phage monoclonal antibodies targeting prostate cancer cells, including those that are hormone refractory. In this report,
we describe the conversion of a panel of those scFvs into full-length human immunoglobulins (IgGs) and show that tumor specificity
is retained. We have further shown that antibodies isolated from a naive phage display library can nevertheless be of high
affinity towards target tumor cells. In addition, full-length IgGs retain the functionality of parental scFvs including the
ability to rapidly enter target cells through receptor-mediated endocytosis and thereby to mediate efficient and specific
intracellular payload delivery to tumor cells. We have used recombinant IgGs to immunoprecipitate target antigens and analyzed
their molecular composition by mass spectrometry. We have identified one target antigen as activated leukocyte cell adhesion
molecule (ALCAM)/MEMD/CD166 and have further studied tissue specificity of this internalizing ALCAM epitope by immunohistochemistry.
Our study shows that cell type-specific internalizing human antibody can be readily identified from a naive phage antibody
display library, characterized with regards to sequence, affinity, tissue specificity, and antigen identity, and modified
genetically and chemically to generate various forms of targeted therapeutics.
Keywords Internalizing phage antibody - Recombinant human IgG1 - Hormone-refractory prostate cancer - Immunoliposomes - ALCAM/MEMD/CD166