Akeda et al. (2018). Vaccine. doi: 10.1016/j.vaccine.2018.02.011
…at 1:8 in dilution buffer (Hanks buffer with Ca2+ and Mg2+, supplemented with 2% BHI medium, 10 mg/ml NAD and 10 mg/ml hemin). H. influenzae (103 bacteria/well), followed by 25 ml of rabbit complement (Pel-Freez) and 25 ml of alamar Blue solution (16% alamar Blue [Trek Diagnostics], 64% dilution buffer, and 2% BHI medium), were added to each of the microtiter plate wells before incubation at 37 C for 6 h in a…
Pel-Freez products cited: Baby Rabbit Complement (31062)
Abstract
Haemophilus influenzae type b (Hib) conjugate vaccines have drastically reduced disease incidence worldwide. Protection against Hib infection has relied on the serum bactericidal activity (SBA) of antibodies to the Hib capsular polysaccharide (polyribosylribitol phosphate). However, licensure usually relies on measuring induction of antibodies to PRP as a surrogate for SBA. In a phase III clinical trial we compared a PRP-conjugate vaccine using the nontoxic diphtheria toxin mutant, CRM<sub>197</sub>, as carrier protein with the licensed tetanus toxoid conjugate when administered subcutaneously as a three dose primary series in Japanese infants. As an addition to the phase III study, we have now evaluated SBA and show PRP-CRM<sub>197</sub> induces higher levels of SBA than PRP-T four weeks after the primary series, with a statistically significant correlation with anti-PRP titers. This data confirms the superior immunogenicity of PRP-CRM<sub>197</sub> compared with PRP-T assessed as SBA following a three-dose primary series by subcutaneous administration. Clinical trial registry: Registered on ClinicalTrials.gov (NCT01379846).