High-throughput antibody screening with high-quality factor nanophotonics and bioprinting
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2024
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| author | Abdollahramezani, Sajjad Omo-Lamai, Darrell Bosman, Gerlof Hemmatyar, Omid Dagli, Sahil Dolia, Varun Chang, Kai Gusken, Nicholas A. Delgado, Hamish C. Boons, Geert-Jan Brongersma, Mark L. Safir, Fareeha Khuri-Yakub, Butrus T. Moradifar, Parivash Dionne, Jennifer A. |
| author_facet | Abdollahramezani, Sajjad Omo-Lamai, Darrell Bosman, Gerlof Hemmatyar, Omid Dagli, Sahil Dolia, Varun Chang, Kai Gusken, Nicholas A. Delgado, Hamish C. Boons, Geert-Jan Brongersma, Mark L. Safir, Fareeha Khuri-Yakub, Butrus T. Moradifar, Parivash Dionne, Jennifer A. |
| contents | Empirical investigation of the quintillion-scale, functionally diverse antibody repertoires that can be generated synthetically or naturally is critical for identifying potential biotherapeutic leads, yet remains burdensome. We present high-throughput nanophotonics- and bioprinter-enabled screening (HT-NaBS), a multiplexed assay for large-scale, sample-efficient, and rapid characterization of antibody libraries. Our platform is built upon independently addressable pixelated nanoantennas exhibiting wavelength-scale mode volumes, high-quality factors (high-Q) exceeding 5000, and pattern densities exceeding one million sensors per square centimeter. Our custom-built acoustic bioprinter enables individual sensor functionalization via the deposition of picoliter droplets from a library of capture antigens at rates up to 25,000 droplets per second. We detect subtle differentiation in the target binding signature through spatially-resolved spectral imaging of hundreds of resonators simultaneously, elucidating antigen-antibody binding kinetic rates, affinity constant, and specificity. We demonstrate HT-NaBS on a panel of antibodies targeting SARS-CoV-2, Influenza A, and Influenza B antigens, with a sub-picomolar limit of detection within 30 minutes. Furthermore, through epitope binning analysis, we demonstrate the competence and diversity of a library of native antibodies targeting functional epitopes on a priority pathogen (H5N1 bird flu) and on glycosylated therapeutic Cetuximab antibodies against epidermal growth factor receptor. With a roadmap to image tens of thousands of sensors simultaneously, this high-throughput, resource-efficient, and label-free platform can rapidly screen for high-affinity and broad epitope coverage, accelerating biotherapeutic discovery and de novo protein design. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2411_18557 |
| institution | arXiv |
| publishDate | 2024 |
| record_format | arxiv |
| spellingShingle | High-throughput antibody screening with high-quality factor nanophotonics and bioprinting Abdollahramezani, Sajjad Omo-Lamai, Darrell Bosman, Gerlof Hemmatyar, Omid Dagli, Sahil Dolia, Varun Chang, Kai Gusken, Nicholas A. Delgado, Hamish C. Boons, Geert-Jan Brongersma, Mark L. Safir, Fareeha Khuri-Yakub, Butrus T. Moradifar, Parivash Dionne, Jennifer A. Optics Biological Physics Empirical investigation of the quintillion-scale, functionally diverse antibody repertoires that can be generated synthetically or naturally is critical for identifying potential biotherapeutic leads, yet remains burdensome. We present high-throughput nanophotonics- and bioprinter-enabled screening (HT-NaBS), a multiplexed assay for large-scale, sample-efficient, and rapid characterization of antibody libraries. Our platform is built upon independently addressable pixelated nanoantennas exhibiting wavelength-scale mode volumes, high-quality factors (high-Q) exceeding 5000, and pattern densities exceeding one million sensors per square centimeter. Our custom-built acoustic bioprinter enables individual sensor functionalization via the deposition of picoliter droplets from a library of capture antigens at rates up to 25,000 droplets per second. We detect subtle differentiation in the target binding signature through spatially-resolved spectral imaging of hundreds of resonators simultaneously, elucidating antigen-antibody binding kinetic rates, affinity constant, and specificity. We demonstrate HT-NaBS on a panel of antibodies targeting SARS-CoV-2, Influenza A, and Influenza B antigens, with a sub-picomolar limit of detection within 30 minutes. Furthermore, through epitope binning analysis, we demonstrate the competence and diversity of a library of native antibodies targeting functional epitopes on a priority pathogen (H5N1 bird flu) and on glycosylated therapeutic Cetuximab antibodies against epidermal growth factor receptor. With a roadmap to image tens of thousands of sensors simultaneously, this high-throughput, resource-efficient, and label-free platform can rapidly screen for high-affinity and broad epitope coverage, accelerating biotherapeutic discovery and de novo protein design. |
| title | High-throughput antibody screening with high-quality factor nanophotonics and bioprinting |
| topic | Optics Biological Physics |
| url | https://arxiv.org/abs/2411.18557 |