Reveille et al. (2024). Vaccine. doi: 10.1016/j.vaccine.2024.126508
…strain lacking pora expression and expressing a hybridized porb (och) [18 ]. see table s3 for a complete description of pc reagents. 3.4 intrinsic sba growth assay igg/igm-depleted human complement (c′) pooled serum (pel-freez validation lot #03642) was screened for endogenous killing activity against ng strains f62, ms11, and fa1090 as previously described [19 ] with modifications. briefly, sialylated or non-sial…
Pel-Freez products cited: Human Complement, Pooled, IgG/IgM Depleted (34010)
Abstract
The recent entry of multiple vaccines targeting Neisseria gonorrhoeae (Ng) into the clinical development pathway has highlighted the need to establish methods of adequately assessing anti-gonococcal immune responses. Serum bactericidal activity (SBA) is utilized as a measurement of efficacy in licensure of meningococcal vaccines, but the importance of functional antibodies in preventing and/or clearing gonococcal infections remains largely unknown. In an effort to elucidate the utility of SBA as an immune correlate of protection, we sought to develop a standardized high-throughput human complement SBA (hSBA) assay for which any strain of interest could be tested under uniform conditions, with minimal screening of complement required. Utilization of IgG/IgM-depleted pooled human complement serum permitted testing of diverse serum resistant and sensitive gonococcal and unencapsulated meningococcal strains when bacteria were cultured to induce lipooligosaccharide sialylation; only one of nine randomly selected lots demonstrated intrinsic toxicity. The hSBA assay was well suited for use with multiple test serum sources, including rabbit, mouse, and human serum samples, suggesting widespread applicability.