Patent ID: 7666627
Filing Date: 2010-02-23
Classification: C07K,C12N

Abstract:
1. A recombinant method for the preparation of an unglycosylated folded C-terminal fragment of a multidomain serine protease of the vertebrate complement cascade, wherein said multidomain serine protease is MASP-1, said method comprising the following steps: i) inserting into a bacterial expression vector a DNA sequence encoding a C-terminal fragment of a vertebrate MASP-1 serine protease and wherein the encoded C-terminal MASP-1 protein fragment consists of the domain structure CCP1-CCP2-SP fused at its amino terminus to a peptide amino acid sequence of either SEQ ID NO:19 or SEQ ID NO:20, ii) transforming a bacterial host cell with said expression vector, iii) inducing the expression of the MASP-1 C-terminal fragment consisting of the domain structure CCP1-CCP2-SP fused at its amino terminus to a peptide of SEQ ID NO:19 or SEQ ID NO:20, iv) isolating inclusion bodies comprising the unglycosylated MASP-1 C-terminal fragment fusion protein of step (iii) consisting of the domain structure CCP1-CCP2-SP fused at its amino terminus to a peptide of SEQ ID NO:19 or SEQ ID NO:20 from the bacterial host cell, v) renaturing the MASP-1 C-terminal fragment fusion protein of step (iv) to provide unglycosylated re-folded MASP-1 C-terminal fragment fusion protein, and vi) purifying the unglycosylated folded MASP-1 C-terminal fragment fusion protein.