Development of SARS-CoV-2 variant protein microarray for profiling humoral immunity in vaccinated subjects

Profiling Humoral Immunity After Mixing and Matching COVID-19 Vaccines Using SARS-CoV-2 Variant Protein Microarrays

“…Since there is no similar high-throughput platform, we focused on the wild-type SARS-CoV-2 and made side-by-side comparisons with plaque reduction assays and surrogate ELISA assays. In our recent study, we demonstrated that the correlations were high, ranging from R-square 0.97-0.98, and proved the reliability of this high-throughput platform ( 11 ). Cell membrane extract from human cell lines has been used for monitoring self-reactive antibodies and the results did not show any significant changes (data not shown).…”

“…For fabrication of the CoVariant protein microarray, the slides were precoated with aldehyde and stored at 4°C as previously described ( 11 , 13 ). Using a contact printer (CapitalBio SmartArrayer 136, China) to print eight variant spike ECDs, seven spike RBDs, two nucleocapsid proteins, 3 control, and cell membrane mix with 30% glycerol ( Table S1 ) in technical triplicate at 4°C cold room.…”

“…To investigate the humoral response in the mixing or matching of vaccines, it is important to adopt a high-throughput platform to measure all the VOCs. We recently developed a SARS-CoV-2 Variant (CoVariant) protein array that included the spike proteins from wild-type, alpha, beta, gamma, and delta variants ( 11 , 12 ). In this study, we expanded the VOCs collection to include omicron and used it for measuring the humoral immunity in the mixing or matching of AZD1222 and mRNA-1273.…”

Recent Progress in Development and Application of DNA, Protein, Peptide, Glycan, Antibody, and Aptamer Microarrays

“…With the advancement of robotic array spotting systems, several thousands of spots can be achieved simultaneously. Several DNA microarrays [ 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 ], glycan microarrays [ 60 , 61 , 62 , 63 , 64 ], peptide microarrays [ 65 , 66 ], and protein microarrays [ 36 , 67 , 68 , 69 , 70 ] are fabricated using commercially available instruments, e.g., Smart Arrayer, OmniGrid contact microarrayer, Gene chip, Bio odyssey, BioRobotics MicroGRID II microarray printer, etc.…”

“…In the second step, the use of a reducing agent, such as sodium borohydride, converts the imine into an amine ( Scheme 1 B). In the case of microarrays, aldehyde-functionalized surfaces (glass and silicon) that are either commercially available or prepared in-house are used [ 69 , 70 ]. Amine-functionalized DNA [ 52 , 54 , 57 , 58 , 59 ] or antibodies with naturally existing amine groups [ 82 ] are immobilized in the above-mentioned two-step process to form DNA or antibody microarrays.…”