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. Author manuscript; available in PMC: 2022 Apr 21.
Published in final edited form as: Adv Exp Med Biol. 2018;1068:89–102. doi: 10.1007/978-981-13-0502-3_8

Figure 5. Screening for ZIka virus neutralizing antibodies using SCAN.

Figure 5.

Peripheral blood cells of Zika infected patients will be isolated. Purified single-cell suspension will be labeled with anti-CD20-FITC and Calcein violet-405 for live cell and plated onto fabricated nanowells. Labeled cells in the nanowells will be imaged for surface markers and locations on the chips. Capture slide coated with anti-human immunoglobulins will be hybridized. Detection antibody mixture containing IgG-AF-488, IgM-AF594, and ZIKV E-AF532 will be added. Micrograph of anti-ZIKV E-secreting B cells will be generated. Individual ZIKV E-secreting B cells will be picked and performed RT/nested PCR for heavy/light chain sequences. Both chains will be cloned into an expression vector and expressed in 293T cell line. Secreted antibodies will be purified and screened for binding and neutralizing activity against ZIKV. AF: Alexa Fluor