Development of High Throughput Antibody Screening Technique with Minimal Sample Requirements

蘇佑晟1*,楊淑君1#,呂明霈2#,楊裕雄1#

1國立陽明交通大學生物科技所, 2國家半導體研究中心

Yu-Sheng Su1*, Suh-Kuan Yong1#, Ming-Pei Lu 2#, Yuh-Shyong Yang 1#

1Department of Biological Science & Technology, National Yang Ming Chiao Tung University,

2Taiwan Semiconductor Research Institute

The urgent need for the efficient screening of antibodies, for screening of novel antibodies for viral detection, vaccine or drug development, can be exemplified by the recent outbreak of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) that causes almost seven million of death worldwide. Current antibodies screening methods involve enzyme immunoassay (EIA) and Immunoprecipitation (IP) that involves laborious procedure, centralized laboratory instruments, and relatively large amount of sample and reagents. In this report, we propose to use atomic force microscopy (AFM) as an alternative technique for high throughput screening novel potential antibodies with minimal sample. AFM could perform the scanning sample in dry or aqueous condition at sub nm precision with single bio-molecule without additional pre-treatment and labeling of the target. SARS-CoV-2 nucleocapsid protein was used as targets in this study. Anti-nucleocapsid protein antibody was immobilized onto the surface of the polysilicon wafer by using 3-aminopropyltriethoxysilane and glutaraldehyde as linker molecules. AFM scanning results revealed that the chip surface topography varied significantly at different stage of the surface modification and antibody-antigen interaction. The statistic result showed that the average height increased approximately one nanometer from individual molecules. The surface roughness also increased following surface treatment with antibody immobilizations and antibody-antigen interaction. In the future, following the integration with high-speed atomic force microscopy, automated system and analysis software, the high-throughput and label-free antibodies screening system could be achieved.