Rapid solid phase extraction device and methods
Inventors
Nelson, William M. • Armantrout, Kyle • Almassian, David R. • de Vazeille, Aymeric Randanne • Fecteau, Tracy Calvin • Kuserk, Colin
Assignees
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Publication Number
US-11590433-B2
Publication Date
2023-02-28
Expiration Date
Abstract
A method and system for solid phase extraction of a compound of interest from a sample matrix using a syringe having a barrel and a plunger, a sorbent for use with the syringe, and a desalting purification column having an end configured to receive liquid from the syringe body.
Core Innovation
The invention relates to a method for extracting a compound of interest from a sample matrix using a syringe having a plunger and a sorbent. A sample matrix is drawn into the syringe and contacted with a sorbent to bind the compound of interest to the sorbent. An elution buffer is drawn into the syringe containing the sorbent, and the elution buffer containing the compound of interest is processed without centrifugation and micropipettes by pushing through the sorbent.
Subsequent to pushing the elution buffer containing the compound of interest through the sorbent, the elution buffer containing the compound of interest is expressed through a desalting column. The desalting step removes contaminants/PCR inhibitors and enables the compound of interest to be provided in a desalted form suitable for downstream analysis such as real-time PCR assays. The approach includes system structures such as a syringe hub with a sorbent membrane and a desalting column connector/exit, including selectable mating via fittings.
The disclosed workflows further include optional steps and variations oriented to polynucleotide-containing samples, including nucleic acids such as DNA. The method includes sample lysis/incubation with a lysis buffer, optional washing, and desalting using a desalting column that can include gel filtration/size-exclusion chromatography media such as Sephadex. Example results described using real-time PCR Ct values versus QIAamp controls support that PCR-ready samples can be obtained while processing larger volumes and enabling point-of-care/remote use.
Claims Coverage
One independent claim is provided in the relevant claims list. The claim defines four main inventive aspects: syringe plunger draw-in of the sample matrix, contacting with a sorbent to bind the compound of interest, drawing an elution buffer into the syringe, and expressing the elution buffer through a desalting column subsequent to pushing through the sorbent.
Syringe plunger draw-in and sorbent binding
drawing a sample matrix into a syringe having a plunger; contacting the sample matrix with a sorbent to bind the compound of interest to the sorbent
Elution buffer draw-in into the syringe containing the sorbent
drawing an elution buffer into the syringe containing the sorbent
Pushing through the sorbent followed by expressing through a desalting column
expressing the elution buffer containing the compound of interest through a desalting column subsequent to pushing the elution buffer containing the compound of interest through the sorbent
The claim coverage centers on a syringe-with-plunger extraction workflow in which a sorbent binds the compound of interest, an elution buffer is drawn into the syringe, and the eluted material is then expressed through a desalting column after being pushed through the sorbent.
Stated Advantages
Eliminates centrifugation and micropipettes.
Removes contaminants/PCR inhibitors during desalting.
Enables PCR-ready samples.
Supports processing larger volumes.
Enables point-of-care/remote use.
Documented Applications
Real-time PCR assay analysis using PCR-ready desalted samples, supported by example Ct comparisons versus QIAamp controls.
Use for polynucleotide-containing samples requiring desalting to remove PCR inhibitors, including nucleic acids such as DNA.
Point-of-care/remote use scenarios.
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