Preparation and characterization of a recombinant humanized single-chain Fv antibody/human interleukin-2 fusion protein directed against the HER-2/neu (c-erbB2) proto-oncogene product, p185
J. Li et al., Preparation and characterization of a recombinant humanized single-chain Fv antibody/human interleukin-2 fusion protein directed against the HER-2/neu (c-erbB2) proto-oncogene product, p185, TUMOR TARG, 4(2), 1999, pp. 105-114
High dose recombinant human interleukin 2 (rhIL-2) therapy has been used in
the treatment of established tumors in both animal models and patients wit
h advanced melanoma or renal carcinoma. However, because high dose rhIL-2 t
herapy causes severe systemic toxicity in normal tissues, its clinical use
has been limited. Therefore, targeting interleukin-2 (IL-2) to the tumor si
te should improve its anti-tumor-immune response and decrease its systemic
toxicity. In this study, we describe the preparation and characterization o
f a recombinant humanized single-chain Fv (sFv) antibody/IL-2 fusion protei
n. This recombinant fusion protein consists of humanized variable heavy (VH
) and light (V-L) domains of monoclonal antibody (mAb) 520C9 directed again
st the human HER-2/neu(c-erbB2) proto-oncogene product p185 and human IL-2.
The fusion protein was stably expressed in baby hamster kidney cells and s
hown to retain the immunostimulatory activities of IL-2 as measured by IL-2
-dependent cell proliferation and cytotoxicity assays. In addition to its I
L-2 activity, this fusion protein also possesses binding specificity agains
t the HER-2/neu(c-erbB2) proto-oncogene product, p185, as determined by enz
yme linked immunosorbent assay (ELISA) using SKOV 3ip1 cells. Taken togethe
r, these findings suggest that this recombinant humanized sFv antibody/IL-2
fusion protein may provide an effective means of targeting therapeutic dos
es of IL-2 to p185 positive tumors without increasing systemic toxicity or
immunogenicity.