Soluble divalent and multivalent heterodimeric analogs of proteins

US6448071B1 · kind B1 · utility

Assignee

• THE JOHNS HOPKINS UNIVERSITY · US

Inventors

• Jonathan Schneck · Silver Spring, US
• Sean O'Herrin · Baltimore, US

Key dates

• Filing date: Jun 3, 1999
• Grant date: Sep 10, 2002
• Expiry date: Jun 3, 2019

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2799/026
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Specificity in immune responses is in part controlled by the selective interaction of T cell receptors with their cognate ligands, peptide/MHC molecules. The discriminating nature of this interaction makes these molecules, in soluble form, good candidates for selectively regulating immune responses. Attempts to exploit soluble analogs of these proteins has been hampered by the intrinsic low avidity of these molecules for their ligands. To increase the avidity of soluble analogs for their cognates to biologically relevant levels, divalent peptide/MHC complexes or T cell receptors (superdimers) were constructed. Using a recombinant DNA strategy, DNA encoding either the MHC class II/peptide or TCR heterodimers was ligated to DNA coding for murine Ig heavy and light chains. These constructs were subsequently expressed in a baculovirus expression system. Enzyme-linked immunosorbant assays (ELISA) specific for the Ig and polymorphic determinants of either the TCR or MHC fraction of the molecule indicated that infected insect cells secreted approximately 1 &mgr;g/ml of soluble, conformationally intact chimeric superdimers. SDS PAGE gel analysis of purified protein showed that expected mol…