All-in-one sample preparation device and method
View Patent ↗Sample preparation device that allows for a complete bind, wash, elute, buffer-exchange and concentration process to be carried out without sample transfer between multiple devices. The device includes a reservoir, a column for holding chromatography media, a holder region for holding a filtration device, and an outlet. The filtration device plugs into the holder region of the centrifugal device, and the assembly can be placed in an optional holder. The assembly, with or without the optional holder, can be placed in a conventional centrifuge tube for centrifugation. The entire bind, wash, elute, buffer exchange and concentration steps can be carried out with the apparatus without any pipette transfers (and the associated sample losses. The sample preparation device also can be used for binding and washing steps, in which case the filtration device is not needed, and for buffer exchange and concentration steps, in which case the media is not needed.
1. A sample preparation device, comprising:
a. a reservoir/exchange member comprising a sample reservoir;
b. a column in fluid communication with said sample reservoir, said column having a holder region and a region having a finned geometry tapering radially from a relatively thick upper portion to a relatively thin lower portion that converges in an axially extending stem having an open bottom;
c. a filter sealingly engaged to said holder region, or capable of being sealing engaged to said holder region, said filter comprising spaced membranes;
d. wherein when said filter is sealingly engaged to said holder region, said region having a finned geometry positions between said spaced membranes.
2. The sample preparation device of claim 1 , wherein said finned region has a uniform wall thickness.
3. The sample preparation device of claim 1 , wherein said column houses chromatography media.
4. The sample preparation device of claim 3 , wherein said chromatography media comprises beads.
5. The sample preparation device of claim 3 , wherein said chromatography media comprises media that bind a member selected from the group consisting of metal chelate, protein A, glutathione and albumin.
6. The sample preparation device of claim 3 , wherein said column houses a frit.
7. The sample preparation device of claim 1 , wherein said spaced membranes define between them a volume, and wherein said region having a finned geometry occupies said volume and is offset from each said membrane.
8. A method of carrying out a buffer exchange, comprising:
a. providing a sample preparation device comprising a reservoir/exchange member comprising a sample reservoir; a column in fluid communication with said sample reservoir, said column having a holder region and a region having a finned geometry tapering radially from a relatively thick upper portion to a relatively thin lower portion that converges in an axially extending stem having an open bottom;
b. sealing engaging a filter device to said holder region;
c. introducing a sample comprising a first buffer into said sample reservoir;
d. concentrating said sample by applying centrifugal force to said sample;
e. introducing a second buffer into said sample reservoir;
f. applying centrifugal force to said sample again; and
g. recovering the resulting sample from said filtration device.
9. A method of purifying a sample, comprising:
a. providing a sample preparation device comprising a reservoir/exchange member comprising a sample reservoir; a column in fluid communication with said sample reservoir, said column having a holder region and a region having a finned geometry tapering radially from a relatively thick upper portion to a relatively thin lower portion that converges in an axially extending stem having an open bottom;
b. introducing media to said column;
c. introducing a sample to said sample reservoir;
d. causing a fraction of said sample to bind to said media;
e. sealing engaging a filter device to said holder region; and
f. eluting said bound fraction from said media.
10. The method of claim 9 , wherein the fraction of sample bound to said media is a sample of interest.
11. The method of claim 10 , wherein said fraction is eluted with an elution buffer, and wherein said sample of interest is collected and concentrated in said filter.
12. The sample preparation device of claim 1 , wherein said axially extending stem is cylindrical.
13. The sample preparation device of claim 1 , wherein said stem tapers radially inwardly towards said open bottom.
14. The sample preparation device of claim 7 , wherein said region having a finned geometry occupies substantially all of said volume.