Cyberscan ph meter

The preliminary studies were conducted to select culture conditions such as temperature, pH, inoculum volume, and medium volume that have an influence on biomass, adenosine, and cordycepin yield in CS, CS1197. Liquid cultures were inoculated with CS1197 and incubated statically for 14 days at temperatures 4, 12, 18, 24, and 28°C to determine the effect of temperature on mycelia growth. The effect of media pH on mycelia growth was estimated by setting different pH values. The pH of the medium was adjusted to 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0 with 1N HCl or 1N NaOH solutions and incubated at 24°C for 14 days. The pH was measured using the Cyberscan pH meter (Eutech Instruments, Singapore). Further, the different volumes of seed culture at 2%, 5%, 10%, 15%, or 20% (volume ratio) were inoculated into a 250-ml flask containing 50 ml medium to study the effect of inoculum volume on mycelia growth, and the growth response of CS1197 to levels of medium volume was investigated by filling 50, 75, 100, 125, 150 or 200 ml medium into a 250-ml Erlenmeyer flask incubated at 24°C for 14 days.

Particle morphology and size were determined by field emission scanning electron microscopy (FE-SEM) at Icon Analytical Equipment Pvt. Ltd., Mumbai, India. Dynamic light scattering (DLS) was further used to determine particle size of the nanovaccine using Zeta Sizer Nano ZS, Worcestershire, UK (Malvern Zetasizer 3000 HS and He/Ne laser at scattering angles of 173° at 25 °C). The Zeta Sizer Nano ZS instrument was also used to determine the surface charge by measuring zeta potential of the nanoparticles. Other physicochemical parameters such as viscosity (Ostwald viscometer), appearance and pH (Cyberscan pH meter, Eutech Instruments, Thermo Fisher Scientific, Inc.) were evaluated. The protocols for content determination of the antigen and immunoadjuvant are described in the supplementary material.