The invention relates to a recombinant human
nerve growth factor purifying method based on a CHO
cell expression
system. The recombinant human
nerve growth factor purifying method based on the CHO
cell expression
system comprises the following steps: 1), centrifugating supernatant, successively removing living cells and
cell fragments, and removing impurities and
bacteria by filtering; 2), applying a supernatant sample to an EMD SO3-(M)
chromatography column, rinsing unbound
protein and impure
protein, eluting and collecting a
target protein peak; 3), concentrating the
protein with
ultrafiltration equipment with a molecular weight cutoff of 3K; and 4), applying the sample to Superdex 75-containing
chromatography column and collecting the
target protein peak, namely a recombinant human
nerve growth factor stoste. According to the CHO cell expression
system and the recombinant human nerve
growth factor purifying method based on the CHO cell expression system, the recombinant human nerve
growth factor with the purity greater than 98% and the
specific activity higher than the 5* 10 to the power of 5 AU / mg can be prepared; compared with other methods, the recombinant human nerve
growth factor purifying method based on the CHO cell expression system has the advantages that the steps are reduced, the
production cycle is greatly shortened, and the
processing capacity and the yield are improved; therefore, the recombinant human nerve growth factor purifying method based on the CHO cell expression system is application for large-scale production.