Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 17, Pages 6529-6539Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c05567
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Funding
- Ministry of Science and Technology [MOST-110-2628-B-006-022, MOST-108-2320-B-006-054-MY2, MOST 110-2731-M-006-001, MOST-109-2327-B-006-005, MOST 111-2321-B-006-009, MOST-110-2622-E-006-030, MOST-110-2327-B006-005, NHRI-110A1-MRCO-02212103, NCKUH-11002018]
- Medical Research Fund of Kaohsiung Armed Forces General Hospital [KAFGH-D108024, KAFGH-D-109042, KAFGH-D110035, KAFGH-D-111033]
- Headquarters of University Advancement at the National Cheng Kung University, Ministry of Education, Taiwan
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This study utilized a multiplexed spike variant protein microarray (SVPM) to analyze the humoral immune response and neutralizing activity in COVID-19 patients, and found that severe patients had stronger inhibition of ACE2 binding to multiple variants.
The disease progression of COVID-19 varies from mild to severe, even death. However, the link between COVID-19 severities and humoral immune specificities is not clear. Here, we developed a multiplexed spike variant protein microarray (SVPM) and utilized it for quantifying neutralizing activity, drug screening, and profiling humoral immunity. First, we demonstrated the competition between antispike antibody and ACE2 on SVPM for measuring the neutralizing activity against multiple spike variants. Next, we collected the serums from healthy subjects and COVID-19 patients with different severities and profile the neutralizing activity as well as antibody isotypes. We identified the inhibition of ACE2 binding was stronger against multiple variants in severe compared to mild/moderate or critical patients. Moreover, the serum IgG against nonstructural protein 3 was elevated in severe but not in mild/moderate and critical cases. Finally, we evaluated two ACE2 inhibitors, Ramipril and Perindopril, and found the dose-dependent inhibition of ACE2 binding to all the spike variants except for B.1.617.3. Together, the SVPM and the assay procedures provide a tool for profiling neutralizing antibodies, antibody isotypes, and reagent specificities.
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