Extended Data Fig. 1: Optimization of the amount of β2GPI nanoparticles, as well as incubation and magnetic separation times for microbial enrichment.
From: One-day rapid sterility test for human-derived biopharmaceuticals
a-b, Capture efficiency based on the amount of β2GPI nanoparticles injected. Varying volumes of β2GPI nanoparticle solution (1.9 × 1010 particles/μL) were mixed with 5 mL of MSC culture medium containing 5 CFU/mL of P. aeruginosa (a) or either 10 CFU/mL of C. albicans (b) (N = 3). The samples were incubated for 60 mins for microbial cell binding and magnetically separated. The resuspended particles were streaked onto TSA plates, overnight cultured, and quantified to calculate the capture efficiency. Above a particle solution volume of 30 μL, the overall efficiency was saturated. c-d, Capture efficiency based on the incubation time for nanoparticle-to-microorganism binding. The enrichment performance was again tested over different incubation times for P. aeruginosa (c) and C. albicans (d), using the optimized β2GPI nanoparticle solution volume of 30 μL (N = 3). The total counts of the microorganisms after the enrichment procedure started to saturate around 15 mins which, in result, was chosen for our final protocols. e-f, Capture efficiency based on the magnetic separation time. Utilizing the optimized nanoparticle solution volume of 30 μL and an incubation time of 15 min, the enrichment performance was reassessed using P. aeruginosa (e) and C. albicans (f) by varying the duration of magnetic separation (N = 3). The capture efficiency plateaued after 5 min, which was established as the optimal time for magnetic isolation. g, Micrograph illustrating the aggregation of β2GPI nanoparticles in solution as a function of magnetic separation time.
