Fig. 6
Nanotrap particle batch validation and batch to batch reproducibility. a Nanotrap particles are incubated with increasing amount of Bb Lysate spiked in 40 mL of urine; Lane 1: Ladder; Lane 2: Positive control Bb Lyme antigen Grade 2 1 ng; Lane 3: Positive control Bb Lyme antigen Grade 2 0.28 ng; Lanes 4–9: eluate of Nanotrap particles incubated with increasing concentrations of Bb antigen in 40 mL of urine: 0; 1.7; 3.5; 7; 15; 30 pg/mL, respectively b Performance comparison of two batches of Nanotrap particles, NT294 and NT306. Lane 1: Ladder; Lane 2: Positive control Bb Lyme antigen Grade 2 60 pg; Lane 3: eluate of Nanotrap particles batch NT296 incubated with 1 pg/mL Bb antigen urine solution (40 mL); Lane 4: eluate of Nanotrap particles batch NT296 incubated with 16 pg/mL Bb antigen urine solution (40 mL); Lane 5–10: eluate of Nanotrap particles batch NT306 incubated with increasing concentrations of Bb antigen urine solution (40 mL) 0; 2; 4; 8; 16; 32, respectively. c Performance comparison of multiple batches of Nanotrap particles (%CV = 9 % and 5 % at 2 pg/mL and 16 pg/mL Bb antigen in 40 mL of urine, respectively). Lane 1: ladder; lane 2: Positive control Bb Lyme antigen Grade 2 16 pg; lane 3: eluate of Nanotrap particles batch NT283 incubated with 40 mL of urine without Bb Lyme antigen Grade 2; lanes 4–5: eluates of Nanotrap particles batch NT283 incubated with 2 and 16 pg/mL Bb Lyme antigen Grade 2 urine solution (40 mL); lanes 6–7: eluates of Nanotrap particles batch NT294 incubated with 2 and 16 pg/mL Bb Lyme antigen Grade 2 urine solution (40 mL); lanes 8–9: eluates of Nanotrap particles batch NT306 incubated with 2 and 16 pg/mL Bb Lyme antigen Grade 2 urine solution (40 mL). d Yield of Nanotrap particle pre-processing is 93 %. Lane 1: ladder; lane 2: Bb antigen 320 pg, lane 3: eluate of Nanotrap particles batch NT283 incubated with 40 mL of urine without Bb Lyme antigen; lane 4: supernatant; lane 5: eluate of Nanotrap particles batch NT283 incubated with 8 pg/mL Bb Lyme antigen Grade 2 urine solution (40 mL)