Abstract
Previously, we established an antibody, termed 102-10, which recognizes insoluble fibrin exclusively, unlike the previously established anti-insoluble fibrin antibodies that also cross-reacted with fibrinogen. We established that the epitope of this antibody is on the β chain that lines an indented structure that becomes exposed only when insoluble fibrin is formed. The amino acid sequence of the epitope is completely conserved from mouse to humans. This study attempted to determine the most suitable insoluble fibrin clone for future diagnostic and therapeutic development. Binding kinetics and properties of antibodies were evaluated by the surface plasmon resonance analysis (SPR) and ELISA among 1101, 99, 443, and 102-10. Immunohistochemical staining for mouse and human pancreatic cancer tissues were also performed. For frozen sections, visually appropriate staining results were observed at an antibody concentration of 1-10 μg/ml, while for paraffin sections, 10 μg/ml was required. From immunohistochemistry and ELISA analyses, clone 99 and clone 1101 showed almost no nonspecific binding in normal pancreatic tissues. Hybridoma production for 1101 yielded more antibodies than that of 99 and demonstrated good long-term stability. It was, therefore, concluded that clone 1101 would be useful for future clinical development as well as basic research.