Expression of immunoglobulin-cytokine fusion proteins in malignant B cells
AssigneeGSF-Forschungszentrum fur Umwelt und Gesundheit GmbH
InventorRalph Mocikat
Life of this patent
Estimating the term…
Abstract
According to the invention, there is provided a vector for the expression of immunoglobulin-cytokine fusion proteins in malignant B cells at least containing operably linked to each other (a) a region of at least 1.5 kb which is homologous to a region of the μ intron or the κ intron and which lacks a functional Cμ or Cκ enhancer or contains a non-functional C82 or Cκ enhancer; (b) at least one DNA sequence encoding a domain of an immunoglobulin or a part thereof; (c) a DNA sequence encoding a cytokine; and (d) a marker gene selectable in eukaryotic B cells and lacking a functional enhancer region wherein the expression of said marker following integration is controlled by the cellular Cμ or Cκ enhancer.
Claims7
Method for the expression of an immunoglobulin-cytokine fusion protein in malignant B cells in vitro, said method comprising:
- (a) introducing into a malignant B cell a vector comprising the following components operably linked together:
- (i) a region of at least 1.5 kb which is homologous to a region of the μ intron or the κ intron,
- (ii) at least one DNA sequence encoding a constant region of an immunoglobulin or a part of the constant region,
- (iii) a DNA sequence encoding a cytokine, and
- (iv) a marker gene which is selectable in eukaryotic B cells and contains a functional enhancer,
- whereby after homologous recombination said immunoglobulin-cytokine fusion protein is expressed, comprising the variable region of the endogenous immunoglobulin of said malignant B cell fused with said constant region of an immunoglobulin or a part of said constant region and said cytokine;
- and
- (b) treating the cells to render them replication-incompetent.
Dependencies are inferred from the claim text.