Fig. 2.

Characteristics of λ phage particles expressing human ASPH protein. (A) The map of the human ASPH sequence containing 758 amino acids (NCBI accession No. NP_004309) compared to the structural map of the murine ASPH sequence containing 725 amino acids (NCBI accession No. NP_001171320). The yellow highlighted sequences represent the homology between mice and humans. The red sequences shows the murine verses human sequences in the λ3 vaccine construct. (B) Representative example of high level ASPH expression in murine BNL derived HCC tumor by IHC using a rabbit polyclonal antibody prepared against human recombinant ASPH protein [18]. The control was rabbit IgG employed as a primary antibody (1000×). (C) Homologous recombination of donor plasmid pVCDcDL3 with recipient phage vector λ-DL1. Only part of the lambda genes are shown. The unique Nhe I and BssH II sites were used to clone ASPH segments into the pVCDcDL1A plasmid. Generated recombinant phages were designated as ASPH construct λ3 which contained the inserted ASPH fragments. The inserts were fused with the GpD capsid protein gene of the λ phage to produce GpD-ASPH fusion construct located on the λ head. Only the diagram of the λ1 construct is shown here. The maps are not drawn to scale.