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It is well documented that antibodies are produced by several antibody-producing cells. However, it is still challenging to synthesize human antibodies in microbial cells. In this study, we genetically engineered yeast Saccharomyces cerevisiae to express two specific antibodies simultaneously using the co-expression strategy. To meet this requirement, we developed a strategy of constructing a yeast strain producing two target proteins in two separate plasmids. Two antibodies were co-expressed in yeast via co-transformation of two corresponding expression vectors, and two antibodies were then co-expressed in the target strain simultaneously. In the expression process of two target proteins, both of them were simultaneously expressed in the cytosol. However, the two antibodies were secreted separately from the cytoplasm by using different signal peptides. It was found that co-expression of two antibodies in the ectodomain of alpha-amylase in yeast strain showed higher production levels of each antibody than the control strain co-expressing single antibody.Q: