Objective To screen GS-CHO cell lines that have high expression of anti-PD-1 antibody, their suitable basal and feed medium, and to analyze the metabolites. Methods We screened from 6 recombinant cell lines and 5 preferred commercial mediums by fed-batchin TPP tube and shake flask. At the same time, we measured glucose, glutamate, glutamine, lactate, and ammonia concentrations. To analyze restricted nutritional factors, we further explored amino acid metabolism in high-yield cell lines. Results With TPP tubes preliminary screening, 122 and 126 cell lines performed best in HyCell CHO medium with titer expression levels of 1.945 g/L and 1.989 g/L, respectively. Further verification in shake flasks showed that PVCD and titer of the 122 cell line in HyCell CHO medium were the highest, which could reached 2.21×10⁷ cells/mL and titer greater than 2 g/L. Meanwhile, it was found that lactic acid metabolism switching was associated with glutamate and alanine metabolism. Amino acid metabolism revealed that essential amino acids such as methionine, isoleucine, and leucine may be the restrictive factors of growth and/or expression for 122 cell line. Conclusion We screened cell line with high expression of anti-PD1 antibody and corresponding optimal medium. Furthermore, metabolism analysis of metabolites and amino acid may provide rational guidance for the subsequent cell culture process development and customized medium optimization.