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Discover how PEPperPRINT Peptide Microarray products have been used in different fields of research.

Non-Neutralizing Antibody Functions Predict Susceptibility to SARS-CoV-2 Infection after mRNA Booster Vaccination

Levy, Shlomia; Trifkovic, Sanja; Mielke, Dieter; Oppenheimer, Hannah; Goodman, Derrick; Ostrovsky, Daniel; Sanfield-Oakley, Sherry A.; Brackett, Caroline; Friedman, Lilach M; Kerkau, Melissa; Webby, Richard; Tomaras, Georgia; Guido, Ferrari; Nesher, Lior; Hertz, Tomer
Previous studies have shown that neutralizing and binding antibody titers are correlates of protection for symptomatic SARS-CoV-2 infection. We previously reported that individuals with low IgG and IgA baseline immune history (BIH) to SARS-CoV-2 variants were at increased risk of symptomatic infection in study of healthcare workers that received 3 or 4 doses of the Pfizer BNT-1262b2 vaccine. We also found that 1-month post-vaccination with the 4th booster dose, individuals with low-BIH mounted significant rises in binding and neutralizing antibody titers, to levels comparable to those of individuals with high-BIH, demonstrating that their increased risk was not due to inability to respond to vaccination. To further study the underlying factors that are associated with increased risk, we conducted a systems serology study of 40 low-BIH and 40 high-BIH individuals across 7 months of follow-up. We found that individuals with low BIH exhibited a significantly higher risk of symptomatic infection (HR=2.691, p=0.0065) and mounted weaker IgA and antibody-dependent cellular cytotoxicity (ADCC) responses compared to high-BIH individuals, particularly against Omicron and Delta variants. Baseline levels of the chemokine CXCL-11 were elevated in the low-BIH group. We then showed that baseline immune profiles can be used to train a prediction model of infection risk across 7 months of follow-up with 76% accuracy. IgA, ADCC and ADCP baseline features were dominant predictors of susceptibility. Our findings suggest that non-neutralizing antibody functions, especially IgA and ADCC, contribute to protection against symptomatic SARS-CoV-2 infection and that serology-guided stratification can enhance discovery of immune correlates of risk informing future vaccine design and deployment strategies.

A Novel Combinatorial Approach to High-Density Peptide Arrays

Beyer, Mario; Block, Ines; König, Kai; Nesterov, Alexander; Fernandez, Simon; Felgenhauer, Thomas; Schirwitz, Christopher; Leibe, Klaus; Bischoff, Ralf F.; Breitling, Frank; Stadler, Volker
Combinatorial synthesis of peptides on solid supports (1), either as spots on cellulose membranes (2) or with split-pool-libraries on polymer beads (3), substantially forwarded research in the field of peptide-protein interactions. Admittedly, these concepts have specific limitations, on one hand the number of synthesizable peptide sequences per area, on the other hand elaborate decoding/encoding strategies, false-positive results and sequence limitations. We recently established a method to produce high-density peptide arrays on microelectronic chips (4). Solid amino acid microparticles were charged by friction and transferred to defined pixel electrodes onto the chip’s surface, where they couple to a functional polymer coating simply upon melting (Fig. 16.1 A-D,F). By applying standard Fmoc chemistry according to Merrifield, peptide array densities of up to 40,000 spots per square centimetre were achieved (Fig. 16.1G). The term Merrifield synthesis describes the consecutive linear coupling and deprotecting of L-amino acids modified with base-labile fluorenylmethoxy (Fmoc) groups at the N-terminus and different acid-sensitive protecting groups at their side chains. Removing side chain protecting groups takes place only once at the very end of each synthesis and generates the natural peptide sequence thereby.

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