Method and materials for isolation of nucleic acid materials

Inventors

Brahmasandra, Sundaresh • Mastronardi, Michelle • Craig, Elizabeth • Carey, Maureen

Assignees

NeumoDx Molecular Inc

Abstract

A method for nucleic acid isolation comprising: receiving a binding moiety solution within a process chamber; mixing the binding moiety solution with a biological sample, within the process chamber, in order to produce a moiety-sample mixture; incubating the moiety-sample mixture during a time window, thereby producing a solution comprising a set of moiety-bound nucleic acid particles and a waste volume; separating the set of moiety-bound nucleic acid particles from the waste volume; washing the set of moiety-bound nucleic acid particles; and releasing a nucleic acid sample from the set of moiety-bound nucleic acid particles. The method preferably utilizes a binding moiety comprising at least one of poly(allylamine) and polypropylenimine tetramine dendrimer, both of which reversibly bind and unbind to nucleic acids based upon environmental pH.

Core Innovation

A method for nucleic acid isolation receives a biological sample and incubates affinity moiety-coated microparticles with the biological sample to reversibly bind nucleic acid material. The affinity moiety-coated microparticles comprise magnetic microparticles bonded to a moiety comprising at least one of Poly(allylamine) and Polypropylenimine tetramine dendrimer, and incubation produces moiety-bound nucleic acid particles within a moiety-sample mixture.

The method occludes a fluidic pathway at a first set of occlusion positions to define a first truncated pathway upon actuating a cam card relative to a first subset of a set of pins corresponding to the first set of occlusion positions. Occluding the fluidic pathway at the first set of occlusion positions defines a fluidic path through a magnetic field and a heating region, and separating comprises delivering the moiety-bound nucleic acid particles into the first truncated pathway and capturing the moiety-bound nucleic acid particles within the magnetic field.

After separating the moiety-bound nucleic acid particles from the moiety-sample mixture, the method releases the nucleic acid material from the moiety of the moiety-bound nucleic acid particles. The document further describes covalent coupling of affinity moieties to magnetic microparticles and performance examples spanning different sample matrices and targets using pH-modulated elution with moieties including PAA and DABAM.

Claims Coverage

The document provides one independent claim directed to a nucleic acid isolation method using reversible moiety-coated magnetic microparticles plus a cam-card/occlusion routed truncated fluidic path through a magnetic field and heating region, followed by release of nucleic acids from the moiety. Dependent claims refine the independent claim with additional target/sample specificity, quantitative constraints, and added fluidic routing.

Across the independent claim and its dependents, the coverage centers on reversible binding of nucleic acids using PAA and/or polypropylenimine tetramine dendrimer moieties on magnetic microparticles, separating and capturing those moiety-bound nucleic acid particles by routing through an occlusion-defined truncated fluidic pathway that passes through a magnetic field and heating region, and releasing nucleic acid material from the moiety after separation.

Stated Advantages

Documented Applications