Antibodies against
severe acute respiratory syndrome coronavirus 2 (
SARS-CoV-2) target multiple
epitopes on different domains of the spike
protein, and other
SARS-CoV-2 proteins. We developed a
SARS-CoV-2 multi-
antigen protein microarray with the
nucleocapsid, spike and its domains (S1, S2), and variants with single (D614G, E484K, N501Y) or double substitutions (N501Y/Deletion69/70), allowing a more detailed high-throughput
analysis of the antibody repertoire following
infection. The assay was demonstrated to be reliable and comparable to
ELISA. We analyzed
antibodies from 18 COVID-19
patients and 12 recovered convalescent
donors. S
IgG level was higher than N
IgG in most of the COVID-19
patients, receptor-binding domain of S1 showed high reactivity, but no
antibodies were detected against heptad repeat domain 2 of S2. Furthermore,
antibodies were detected against S variants with single and double substitutions in COVID-19
patients who were infected with
SARS-CoV-2 early in the
pandemic. Here we demonstrated that
SARS-CoV-2 multi-
antigen protein microarray is a powerful tool for detailed characterization of
antibody responses, with potential utility in
understanding the
disease progress and assessing current
vaccines and
therapies against evolving
SARS-CoV-2.