Integrated membrane for preservation of biomolecules
Inventors
Menon, Naresh • Bui, Peter • Tan, Cheryl
Assignees
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Publication Number
US-9933343-B2
Publication Date
2018-04-03
Expiration Date
Abstract
The apparatus of the present invention comprises an integrated membrane for separation and preservation of biomolecules. Biomolecules, such as those in blood, can be separated and preserved. The apparatus includes a housing having a matrix portion, and a fluid collection portion disposed within an inner cavity. The housing further includes an aperture permitting access to the inner cavity. The apparatus also includes a matrix disposed within the matrix portion. The matrix includes a first layer for collecting cells from the fluid, a second layer for protein adsorption, and a third layer for nucleic acid adsorption. Fluid enters the housing through the aperture, passes through the matrix, and into the fluid collection portion. Cells from the fluid in a layer of the matrix are collected; adsorbing protein from the fluid in a layer of the matrix; and adsorbing nucleic acid from the fluid in a layer of the matrix.
Core Innovation
The invention relates to an apparatus for separating and preserving biomolecules of a biological fluid using an assembly having sides forming a hollow shape with an opening at each of two ends. Matrix layers are positioned within the hollow shape between the ends, and the sides are reversibly and hydrophobically sealed with the matrix layers to actuate a flow of the biological fluid in a direction from a first matrix layer toward a second matrix layer and, in some embodiments, toward a third matrix layer.
In the described flow-through arrangement, the first matrix layer is configured to receive the biological fluid and a lysis buffer, and includes a first membrane layer configured to trap proteins and to allow nucleic acids to flow through onto the next matrix layer. The next matrix layer is configured to selectively bind nucleic acids and to separate from the assembly in a direction away from the first matrix layer.
In embodiments with three matrix layers, the first and second membrane layers trap proteins while allowing proteins and nucleic acids to flow onto subsequent layers until nucleic acids are selectively bound on the third matrix layer. The matrix layers and membranes are configured to enable moiety-specific separation while preserving biomolecules, including nucleic acids, and embodiments include lines of weakness forming at least two sections to allow independent removal of at least one section.
Claims Coverage
The document contains two independent claims, each directed to an apparatus for separating and preserving biomolecules from a biological fluid using reversibly and hydrophobically sealed, flow-through membrane matrix layers with selective nucleic-acid capture. Claim coverage focuses on two-layer and three-layer architectures, including structural and material refinements to enable independent removal of membrane portions and specified membrane properties.
Two-layer sealed flow-through nucleic-acid selective separation
An apparatus for separating and preserving biomolecules of a biological fluid with an assembly having sides forming a hollow shape with openings at two ends; two matrix layers positioned within the hollow shape, the first matrix layer receiving the biological fluid and a lysis buffer and including a first membrane layer configured to trap proteins and allow nucleic acids to flow through; and a second matrix layer including a second membrane layer configured to selectively bind nucleic acids, with the sides reversibly and hydrophobically sealed to actuate flow from the first matrix layer to the second and with the second matrix layer configured to separate in a direction away from the first matrix layer.
Three-layer sequential protein trapping and nucleic-acid selective binding
An apparatus for separating and preserving biomolecules of a biological fluid with an assembly having sides forming a hollow shape with openings at two ends; three matrix layers positioned within the hollow shape, the first matrix layer receiving the biological fluid and a lysis buffer and including a first membrane layer configured to trap proteins and allow proteins and nucleic acids to flow through onto the second matrix layer; the second matrix layer including a second membrane layer configured to trap proteins and allow nucleic acids to flow through onto the third matrix layer; and the third matrix layer including a third membrane layer configured to selectively bind nucleic acids, with the sides reversibly and hydrophobically sealed to actuate flow from the first matrix layer to the second matrix layer to the third matrix layer and with the second and/or third matrix layers configured to independently separate in a direction away from the first matrix layer.
Across the independent claims, the inventive concept is a sealed, hydrophobic assembly that actuates controlled flow through sequential membrane matrix layers where early membranes trap proteins and allow nucleic acids to pass, and a later membrane layer selectively binds nucleic acids. The two-layer and three-layer versions further emphasize separability of the nucleic-acid-binding matrix layer(s), including independent separation where structural features such as lines of weakness divide layers into sections.
Stated Advantages
Eliminating protein denaturants/solvent steps interfering with downstream PCR.
Rapid processing.
Minimal sample volumes.
Reduction of contamination/cross-contamination.
Compatibility with ambient storage.
Documented Applications
Separating and preserving biomolecules, including moiety-specific separation and ambient preservation of biomolecules from blood.
Downstream analysis including PCR referenced as being affected by protein denaturants/solvent steps.
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