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. 2014 May 15;9(5):e95137. doi: 10.1371/journal.pone.0095137

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© 2014 Nakao et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Shown are results of the assays using crude MV preparations from the following five different species: E. coli KP7600, N. meningitidis H44/76, P. aeruginosa PAO1 Holloway, C. perfringens strain 13, and P. gingivalis ATCC 33277. (A-D) Crude MVs from a single strain was incubated with the SB-Epoxy. Unbound components were collected in five washes and bound components were eventually eluted by SDS-PAGE loading buffer. Lanes denoted “1” are the starting material from conventional purification (Crude). Lanes denoted “2” are the first unbound fractions (Unbound, 1^st). Lanes denoted “3” are the fifth unbound fractions (Unbound, 5^th). Lanes denoted “4” are the elution fractions (Elu). (A: E. coli KP7600, B: N. meningitidis H44/76, C: P. aeruginosa PAO1-Tokai, D: C. perfringens strain 13). (E) Crude MVs from the five species were mixed with SB-Epoxy. The selective elimination of P. gingivalis MVs from the mixture is shown. Lanes 5 &12: the input of mixtures of crude MVs (Crude mixture). Lanes 6 & 13: the first unbound fractions (Unbound, 1^st). Lanes 12 & 13: five-fold diluted samples of lanes 5 & 6, respectively. Lanes 7, 8, & 9: the third unbound fraction (Unbound, 3^rd), the fifth unbound fraction (Unbound, 5^th), and the elution fraction (Elu-Mix) from the mixture of four different MV preparations, respectively. Lane 10: the molecular weight marker. Lane 11: the crude MVs of P. gingivalis (Pg, Crude). Lane 14: the elution fraction from crude MVs of P. gingivalis only (Elu-Pg). The same volume (10 µl) was applied in each well of each PAGE gel. P. gingivalis FimA is denoted by an asterisk (*). N. meningitidis PorB is denoted by a dagger (†).