Rapid selection of single cells with high antibody production rates by microwell array |
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Authors: | Park Sunyoung Han Jieun Kim Wanjung Lee Gyun Min Kim Hak-Sung |
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Institution: | a Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea b Graduate school of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea |
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Abstract: | Selection of single cells capable of producing target proteins at high rates is crucial for the development of protein manufacturing processes. Here, we present the rapid selection of single cells producing immunoglobulin antibodies at high specific rates by microwell array and microengraving. Chinese hamster ovary (CHO) cells secreting chimeric antibodies were deposited in a microwell array in a manner such that each microwell contained a single cell. Secreted antibodies in the microwells were transferred onto a glass slide by microengraving, followed by interrogation using fluorescence-based immunoassay. Single cells displaying high signal intensities were selected, retrieved, and clonally expanded to assess their specific antibody production rates. Three successive rounds of the process resulted in the selection of single cells showing significantly increased antibody production rates. The present approach can be applied to the selection of single cells for producing other therapeutic proteins in a high-throughput manner. |
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Keywords: | Single cell Antibody Microwells Microengraving CHO cell |
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