Generating single-domain antibodies against fibronectin splice variants

Abstract

Here, I describe the process of generating single-domain antibodies which bind to splice variants of fibronectin containing EIIIA or EIIIB. Alpacas were immunized with either purified antigen cocktails, or from the ECM of tumor samples, and antibody libraries were generated. Using these libraries to pan against, I selected for VHH which bind to EIIIA and EIIIB. Since these splice variants are upregulated in tumor angiogenesis and are rarely seen elsewhere in adult tissues, antibodies targeting EIIIA or EIIIB may be of use for imaging tumors and metastases.