Abstract
Selective high-affinity ligands (SHALs) were selected as substitutes for monoclonal antibodies (mAbs) to deliver radioisotopes to malignant tumors. Because a SHAL (5 KD) is considerably smaller in comparison to an antibody (150 KD), a significant therapeutic index (TI) enhancement for radioimmunotherapy (RIT) is anticipated. The antibody-antigen (Ab-Ag) model system chosen for the development of SHALs consists of Lym-1, a MAb with proven selectivity in non-Hodgkin's lymphoma (NHL) patients and its well-characterized Ag, the β subunit of HLA DR10. Whereas Lym-1 is readily available, the β subunit of HLA-DR10 is not. Native, heterodimeric (α and β subunits) HLA-DR10 can be purified from Raji cells, which are known to overexpress this Ag. Inconsistent homogeneity between preparations of HLA-DR10 solubilized in the presence of detergents prompted us to express a recombinant form of the β subunit of HLA-DR10 in Escherichia coli. Negligible production yields (≤50 μg/L) were achieved by the expression of the full-length protein in a soluble form. By contrast, yields of 240 mg/L were obtained by expressing only the extracellular domain (ED) of the β subunit of HLA-DR10 in an insoluble form (inclusion bodies). The recovery yield of refolded protein was 75%. Circular dichroism (CD) and Lym-1 binding studies indicated that the recombinant ED of the β subunit of HLA-DR10 was properly folded. Therefore, this recombinant protein can be used as a surrogate for native heterodimeric HLA DR10 for the in vitro selection of SHALs and related targeting molecules.
Original language | English (US) |
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Pages (from-to) | 531-542 |
Number of pages | 12 |
Journal | Cancer Biotherapy and Radiopharmaceuticals |
Volume | 22 |
Issue number | 4 |
DOIs | |
State | Published - Aug 2007 |
Keywords
- Antibody
- HLA-DR
- Lymphoma
- Radioimmunotherapy
- Radioisotope
ASJC Scopus subject areas
- Oncology
- Radiology Nuclear Medicine and imaging
- Pharmacology
- Cancer Research