Journal: Biomarker Research

Article Title: Selective isolation of extracellular vesicles from minimally processed human plasma as a translational strategy for liquid biopsies

doi: 10.1186/s40364-022-00404-1

Figure Lengend Snippet: Magnetic beads (Miltenyi Biotec)

Article Snippet: To eliminate antibodies in excess, antibody-coated streptavidin beads were loaded in magnetized pre-equilibrated MACS μColumns (Miltenyi Biotec), washed 3x with 200 μL of PBS-Tween20 0,1% v/v and 2x with 400 μL of PBS.

Techniques: Magnetic Beads, Isolation

Journal: Biomarker Research

Article Title: Selective isolation of extracellular vesicles from minimally processed human plasma as a translational strategy for liquid biopsies

doi: 10.1186/s40364-022-00404-1

Figure Lengend Snippet: Differential recovery of EV subpopulations in simple and complex matrices, between streptavidin-coated and covalently-conjugated beads. ( A ) CFSE-stained HEK293 EVs were isolated from PBS-BSA with MACS-STV and MACS. CD9 was the IP target and ISO/CD61 were negative controls. Recovery was plotted as % of input, obtained using fluorescent signals of samples and input. Fluorescence of IP flow-throughs (FT) allowed for an indirect calculation of recovery (yellow), whereas fluorescence on beads provided a direct recovery measure (blue). Specificity represents the difference (in percentage) between target and negative control signal. ( B ) Cryo-TEM images of triple-coated MACS beads (1), HEK293 EVs (2) and the IP complex formed between both in PBS-BSA (3). Respective scale bars are shown on the top right corner of each image. ( C ) S/N ratios of CFSE-stained HEK293 EVs recovered from plasma (donor 6) on MACS-STV and MACS. The IP target was CD9 and the negative controls were ISO/CD61. ( D ) After RNA isolation from MACS CD9 and CD61 samples obtained in (C), GAPDH and CA9 mRNA copies were measured by ddPCR. ( E ) S/N ratios of CFSE-stained HEK293 EVs recovered from plasma (donor 6) on MACS beads, coated with anti-CD9 and anti-PE antibodies. IP specificity was 78%

Article Snippet: To eliminate antibodies in excess, antibody-coated streptavidin beads were loaded in magnetized pre-equilibrated MACS μColumns (Miltenyi Biotec), washed 3x with 200 μL of PBS-Tween20 0,1% v/v and 2x with 400 μL of PBS.

Techniques: Staining, Isolation, Fluorescence, Negative Control