Stability Analysis of Engineering Cell Line for Production of Long-acting Growth Hormone
The rhGH-Fcfusion protein(AK2017)is a long-acting growth hormone drug for the treatment of growth hormone deficiency.The half-life of the drug is prolonged by fusion of Fc protein at the C-terminus of rhGH,so as to achieve the purpose of long-acting efficacy.The establishment of a stable CHO cell line suitable for production is the prerequisite for the commercial production of rhGH-Fc fusion protein,and it is also an important problem in the development faced by all recombinant protein drugs.To analyze the stability of engineering cell line CHO-rhGH-Fc for production of rhGH-Fc fusion protein,the stability of the engineered cell line CHO-rhGH-Fc for production was evaluated from three aspects of monoclonal origin of cell line screening,passage stability and quality consistency of three batches samples under GMP conditions.The results showed that the monoclonality of CHO-rhGH-Fc cell line for production was about 96.7％,and cell morphology and multiplication rate during passage were consistent with those of the working cell bank before passage.The rhGH-Fc fusion protein titer in supernatant and copy number of target gene in G54 generation were 78％and 87.8％of those in G8 generation,respectively.The growth trend of different generation cells was stable during the whole culture process,and the nucleotide sequence of the target gene was consistent with the original theoretical sequence.Furthermore,during three consecutive batches of 200 L bioreactor scale cultures under GMP conditions,cells growth and metabolic parameters were similar.The purity,post-translational modifications and relative biological activity of three consecutive batches of purified rhGH-Fc fusion protein samples were essentially identical.The results shows that CHO-rhGH-Fc cell line for the production of long-acting growth hormone have good passaging stability and inter-batch stability,and the cells remained identical during the culture cycle of commercial production scale,which could meet the needs of commercial production.
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