Amgen Research (munich) Gmbh

Micromet Ag

Silke Mittelstrass

Tobias Raum

Andreas Wolf

Eva Krinner

Julia Henckel

Patrick Hoffmann

Jens Hennecke

The invention relates to methods of preparing antibody fragments. The invention further relates to antibody fragments prepared by these methods. The invention further relates to antibody variable regions comprised in antibody fragments producible by these methods.

Preparation of scFv antibody fragments

A composite polypeptide, said composite polypeptide comprising a desired polypeptide and an expression enhancing domain ('EED'), said EED comprising first and second cysteine amino acid residues Cys1 and Cys2, respectively, Cys1 being located closer to the N-terminus of the composite polypeptide molecule than Cys2, wherein Cys1 and Cys2 are separated by a polypeptide linker, said linker&#x2014;being free of cysteine and proline;&#x2014;defining a length sufficient to allow Cys1 and Cys2 to engage in an intramolecular disulfide bond with one another; and&#x2014;having a flexible polypeptide conformation essentially free of secondary polypeptide structure in aqueous solution, wherein at least one of Cys1 and Cys2 is derivatized with a derivatization moiety.

Expression-enhanced polypeptides

The invention relates to methods of preparing antibody fragments. The invention further relates to antibody fragments prepared by said methods. The invention further relates to antibody variable regions comprised in antibody fragments producible by said methods.

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Munich, Germany

Silke Mittelstrass