Method for facilitating extraction of a fraction from a biological sample

Patent metadata

Field	Value
Publication number	US-10640530-B2
Application number	US-201815891978-A
Country	US
Kind code	B2
Filing date	Feb 8, 2018
Priority date	Dec 15, 2011
Publication date	May 5, 2020
Grant date	May 5, 2020

Abstract

A method is provided for facilitating extraction of a fraction from a biological sample. The biological sample includes non-desired material and a fraction-bound solid phase substrate. The method includes the steps of capturing the fraction-bound solid phase substrate and bringing an isolation buffer and the fraction-bound solid phase substrate into contact to purify the captured fraction-bound solid phase substrate.

First claim

We claim:

1. A method for facilitating extraction of a fraction from a biological sample, the biological sample including non-desired material and a fraction-bound solid phase substrate, the method comprising the steps of:
positioning at least a portion of a plunger within a passageway extending along axis through a barrel of a syringe, the plunger being slidable along the axis and including:
an elongated stem having a first end received in the barrel; and
a resilient plunger head extending radially from first end of the stem and terminating at a radially outer edge, the radially outer edge of the plunger head forming an airtight seal with an inner surface of the barrel;

sliding the plunger along the axis of the barrel in a first direction such that the plunger head is positioned adjacent an opening in a first end of barrel;
positioning the opening in the first end of the barrel in fluid communication with an isolation buffer;
sliding the plunger along the axis of the barrel in a second direction, opposite to the first direction, for a first user-selected distance so as to draw the isolation buffer into the passageway in the barrel through the opening;
positioning the opening in the first end of the barrel in fluid communication with the biological sample;
sliding the plunger along the axis of the barrel in a second direction for a second user-selected distance so as to draw the biological sample into the passageway in the barrel through the opening;
positioning a magnet adjacent to an outer surface of the barrel in proximity to the first end of the barrel such that a magnetic field generated by the magnet captures the fraction-bound solid phase substrate at a fixed location within the passageway through the barrel of the syringe;
sliding the plunger along the axis of the barrel in the first direction so as to urge the biological sample from the passageway in the barrel through the opening while maintaining the captured fraction-bound solid phase substrate at the fixed location;
sliding the plunger along the axis of the barrel in the first direction such that at least a portion of the isolation buffer contacts the captured fraction-bound solid phase substrate at the fixed location and flows out of the passageway in the barrel through the opening;
removing the magnet from adjacent to an outer surface of the barrel such that the fraction-bound solid phase substrate is no longer captured at the fixed location within the passageway through the barrel of the syringe; and
sliding the plunger along the axis of the barrel in the first direction such that the fraction-bound solid phase substrate is urged from the passageway in the barrel through the opening;

wherein:
the isolation buffer and the biological sample are immiscible; and
prior to the step of positioning the opening in the first end of the barrel in fluid communication with the isolation buffer including the additional steps of:
positioning the opening in the first end of the barrel in fluid communication with an elusion buffer; and
sliding the plunger along the axis of the barrel in the second direction for a third user-selected distance so as to draw the elusion buffer into the passageway in the barrel through the opening.

2. The method of claim 1 wherein prior to the step of removing the magnet from adjacent to the outer surface of the barrel, comprising the additional step of sliding the plunger along the axis of the barrel in the first direction such that at least a portion of the elusion buffer contacts the captured fraction-bound solid phase substrate at the fixed location.

3. The method of claim 1 wherein the elusion buffer is urged from the passageway in the barrel through the opening as the fraction-bound solid phase substrate is urged from the passageway in the barrel through the opening.

4. The method of claim 1 wherein the elusion buffer is immiscible with the isolation buffer and the biological sample.

5. A method for facilitating extraction of a fraction from a biological sample, the biological sample including non-desired material and a fraction-bound solid phase substrate, the method comprising the steps of:
positioning at least a portion of a plunger within a passageway extending along axis through a barrel of a syringe, the plunger being slidable along the axis and including:
an elongated stem having a first end received in the barrel; and
a resilient plunger head extending radially from first end of the stem and terminating at a radially outer edge, the radially outer edge of the plunger head forming an airtight seal with an inner surface of the barrel;

sliding the plunger along the axis of the barrel in a first direction such that the plunger head is positioned adjacent an opening in a first end of the barrel;
positioning the opening in the first end of the barrel in fluid communication with an isolation buffer;
sliding the plunger along the axis of the barrel in a second direction, opposite to the first direction, for a first user-selected distance so as to draw the isolation buffer into the passageway in the barrel through the opening;
positioning the opening in the first end of the barrel in fluid communication with the biological sample;
sliding the plunger along the axis of the barrel in a second direction for a second user-selected distance, so as to draw the biological sample into the passageway in the barrel through the opening;
positioning a magnet adjacent to an outer surface of the barrel in proximity to the first end of the barrel such that a magnetic field generated by the magnet captures the fraction-bound solid phase substrate at a fixed location within the passageway through the barrel of the syringe;
sliding the plunger along the axis of the barrel in the first direction for a third distance to urge the biological sample and at least a portion of isolation buffer from the passageway in the barrel through the opening and the captured fraction-bound solid phase substrate are maintained in the passageway in the barrel at the fixed location;
removing the magnet from adjacent to an outer surface of the barrel such that the fraction-bound solid phase substrate is no longer captured at the fixed location within the passageway through the barrel of the syringe; and
sliding the plunger along the axis of the barrel in the first direction such that the fraction-bound solid phase substrate is urged from the passageway in the barrel through the opening;

wherein prior to the step of positioning the opening in the first end of the barrel in fluid communication with the isolation buffer including the additional steps of:
positioning the opening in the first end of the barrel in fluid communication with an elusion buffer; and
sliding the plunger along the axis of the barrel in the second direction for a third user-selected distance so as to draw the elusion buffer into the passageway in the barrel through the opening.

6. The method of claim 5 wherein prior to the step of removing the magnet from adjacent to the outer surface of the barrel, comprising the additional step of sliding the plunger along the axis of the barrel in the first direction such that at least a portion of the elusion buffer contacts the captured fraction-bound solid phase substrate at the fixed location.

7. The method of claim 5 wherein the elusion buffer is urged from the passageway in the barrel through the opening as the fraction-bound solid phase substrate is urged from the passageway in the barrel through the opening.

8. The method of claim 7 wherein the elusion buffer, the isolation buffer and the biological sample are immiscible.

9. A method for facilitating extraction of a fraction from a biological sample, the biological sample inc

Assignees

• Wisconsin Alumni Res Found

Inventors

• Beebe David
• Strotman Lindsay
• Berry Scott M

Classifications

• C07K1/145

  by extraction or solubilisation · CPC title

• C07K1/14 Primary

  Extraction; Separation; Purification · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title