Protein with Activity 1——MCSF1 (Colony Stimulating Factor 1, Macrophage)

Generally, protein could be used as immunogen based ontheir amino acid sequence. Moreover, protein will possess specific biologicalfunctions while they get spatial conformation, and various kinds of functionsof different proteins will lead to physical functions of organism. So, theresearch of active protein is a hot spot of biological and medical researchfield. As researchers need to use biological active proteins, but natural onesare difficult to obtain, so, most of the native proteins used are generatedform genetic engineering..

There are several steps and expression methods to getproteins by genetic engineering. But, it is necessary to select gene sequence,expression system and activity test method. In the research processes ofCloud-Clone Corp., lots of active proteins are developed. For instance, whiledeveloping the protein of mouse MCSF (colony stimulating factor 1, macrophage),the sequence was analyzed at first, combined with the analysis of structure,sequence of Lys33~Ser204, which is a part of extracellular domain, was chosen forrecombinant protein. And E.coli expression system was used for proteinexpression. As E.coli expressed proteins contain endotoxin, so,experiments were performed to exclude endotoxin after the protein was purified.The endotoxin level is less than 1.0EU/μgfor end-product.

It is know that colonystimulating factor have great impacts on proliferation and differentiation ofmononuclear phagocyte, especially macrophage and monocyte, so, in order toverify whether the recombinant mouse MCSF have bio-activity, we used BMDC (bonemarrow derived dendritic cell) in the test. After cultured in culture mediumsupplemented with the recombinant protein, cells were observed by microscopy.It is found that the number of cells in MCSF cultured group is significantlyincreased comparing with negative control (Fig. 1). The result displayed thatrecombinant mouse MCSF (RPA090Mu01) have bio-activity of promoting the proliferationof BMDC.

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A.BMDCcultured in DMEM Media (Negative Control).B.BMDC cultured in DMEM Media supplemented with MCSF.