Anti‐S2 Protection in COVID‐19 Infection and SARS‐CoV‐2 Spike Vaccination.

R4586 --> 524.8 --> The overall goal of this project is to define the magnitude, quality, and duration of the primary immune response elicited against SARS‐CoV‐2 Spike by measuring domain‐specific antibody abundance and binding characteristics in plasmas after infection and vaccination. This investigation has enabled initiation of the screening of convalescent plasma polyclonal antibody (pAb) abundance and specificity through the IMPACC (Immunophenotyping Assessment in a COVID‐19 Cohort) at Drexel U College of Medicine (DUCOM) in collaboration with Tower Health Hospitals. We measured the active concentration of pAbs specific for RBD, S1 and S2 domains using SPR (surface plasmon resonance) molecular interaction analysis. By adopting a kinetic format, a complementary SPR analysis step was optimized to determine the binding rates and affinities of elicited antibodies targeting each domain of the Spike using the same plasma dilution aliquot. Most importantly, we found that the abundance of S2 reactive antibodies was comparable to that of anti‐S1 and RBD in convalescent plasmas. Plasmas obtained up to 6 months post‐vaccination are also becoming available through the TTC (Vaccination TetraCore cohort) assessment at DUCOM, and screening for these has demonstrated that anti‐S2 pAbs are also elicited, though intriguingly in lower abundance than after infection. To assess the importance anti‐S2 antibodies from convalescent plasmas, we purified anti‐S2 fractions by an SPR‐based microaffinity method and used the recovered antibodies in pseudovirus infection inhibition assays of ACE2 expressing cells to measure neutralization activity. Evidence for sustained generation of S2 antibodies up to 6 months post‐infection and occurrence of neutralizing anti‐S2 pAbs has begun to emerge with the possibility that antibodies targeting the S2 domain of the SARS‐CoV‐2 spike protein complex could provide pan‐coronavirus protection against COVID‐19, emerging variants, and other coronaviruses with conserved spike structures. Targeting the more conserved fusion machinery in the virus spike ultimately can lead to therapeutic antibodies or small molecule inhibitors effective on escape variants that occur mainly in S1 as well as other coronaviruses. [ABSTRACT FROM AUTHOR]