Expression of Hirudin Fusion Proteins in Mammalian Cells

Expression of hirudin fusion proteins in mammalian cells: a strategy for prevention of intravascular thrombosis.

BACKGROUND Intravascular thrombosis occurs in disorders of diverse pathogeneses, including allograft and xenograft rejection. In this in vitro study, we describe an approach for tethering the specific thrombin inhibitor hirudin to plasma membranes as part of a genetic strategy for regulating intravascular coagulation. METHODS AND RESULTS An HLA class I leader sequence was fused with hirudin l...

Transient production of scFv-Fc fusion proteins in mammalian cells

Single chain Fv (scFv) antibody fragments fused to the human immunoglobulin G (IgG) Fc moiety obtain IgG-like properties but nevertheless they are still encoded by a single gene. Since transient production of scFv-Fc proteins in mammalian can easily achieve milligram amounts, this antibody format is particularly suitable for many research applications. Here, we describe two protocols for the tr...

pLEF, a novel vector for expression of glutathione S-transferase fusion proteins in mammalian cells.

An expression vector, pLEF, has been used to produce the intracellular domain (IC) of the human CD95 (Fas/APO-1) apoptosis receptor as a glutathione S-transferase (GST) fusion protein in murine L929 fibroblasts. GST::CD95IC was affinity-purified in a single step using glutathione-Sepharose. Purification of GST::CD95IC from 32P-labelled L929 cells and cleavage with thrombin revealed that CD95IC ...

Expression of Biologically Active Recombinant B-Domain-Deleted Human Factor VIII in Mammalian Cells

Plasticity of HP1 proteins in mammalian cells.

We have compared the distribution of endogenous heterochromatin protein 1 (HP1) proteins (alpha, beta and gamma) in different epithelial lines, pluripotent stem cells and embryonic fibroblasts. In parallel, we have interrogated assembly and dynamics of newly expressed HP1-GFP proteins in cells lacking both HP1alpha and HP1beta alleles, blocked at the G1-S boundary, or cultured in the presence o...