Anaphore, Inc.

Hoffmann-la Roche Inc.

Denzyme Aps

Michael Etzerodt

Thor Las Holtet

Niels Jonas Heilskov Graversen

Hans Christian Thøgersen

Jette Sandholm Kastrup

Bettina Bryde Nielsen

Ingrid Kjøller Larsen

Hans Christian Thogersen

Ingrid Kjoller Larsen

Thor Lns Holtet

Hans Christian Tho Slashedgersen

Christian Tho Slashedgersen

The present invention relates to the design of trimeric polypeptides using polypeptide structural elements derived from the tetranectin protein family, and their use in rational de novo design and production of multi-functional molecules including the application of the multi-functional molecules in protein library technology, such as phage display technology, diagnostic and therapeutic systems, such as human gene therapy and imaging. The trimeric polypeptides being constructed as a monomer polypeptide construct comprising at least one tetranectin trimerising structural element (TTSE) which is covalently linked to at least one heterologous moiety, said TTSE being capable of forming a stable complex with two other TTSEs; or as an oligomer which is comprised of two monomer polypeptide constructs as mentioned above, and which comprises three TTSEs or a multiplum of three TTSEs, or which is comprised of three monomer polypeptide constructs.

Trimerising module

The present invention provides novel polypeptides having the scaffold structure of a C-type lectin-like domain (CTLD) and a randomized loop region for specifically binding a variety of target compounds and also provides nucleic acids encoding the polypeptides. The present invention further provides combinatorial CTLD libraries, methods for constructing the libraries, and methods for screening the libraries to identify and isolate the novel CTLD polypeptides. Specifically, the invention provides libraries of nucleic acids encoding polypeptides having a scaffold CTLD with a randomized loop region, as well as nucleic acid sequences, vectors, and methods for preparing and expressing the libraries. Exemplary nucleic acids useful in the combinatorial libraries are derived from tetranectin and other proteins having a CTLD.

Combinatorial libraries of proteins having the scaffold structure of C-type lectin-like domains

A novel, generally applicable method for producing correctly folded proteins from a mixture of misfolded proteins, e.g. bacterial inclusion-body aggregates. A major new aspect of the method is that over-all efficiency is achieved by subjecting proteins to a time-sequence of multiple denaturation-renaturation cycles, resulting in gradual accumulation of the correctly folded protein. The method has proven efficient for a variety of recombinant proteins. Also provided are novel encrypted recognition sites for bovine coagulation factor X.sub.a. The encrypted recognition sites described may be activated in vitro by controlled oxidation or by reversible derivatization of cysteine residues and thereby generate new cleavage sites for factor X.sub.a. Two new recombinant serine protease exhibiting narrow substrate specificity for factor X.sub.a recognition sites are also provided. They may replace natural coagulation factor X.sub.a for cleavage of chimeric proteins.

Iterative method of at least five cycles for the refolding of proteins

Interative method of at least three cycles for the refolding of proteins

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Hinnerup, Denmark

Michael Etzerodt