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US 12,258,587 B2Granted
Compositions and methods for internal controls of microvesicle isolations
AssigneeExosome Diagnostics, Inc.
InventorsGeorg Stoll, Daniel Enderle, Mikkel Noerholm, Johan Karl Olov Skog
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Abstract
The present disclosure provides compositions, methods and kits for internal controls of microvesicle isolations. The compositions, methods and kits can comprise enveloped viruses, including, but not limited to, inactive mouse hepatitis virus (MHV).
Claims19
A method comprising:
- (a) adding a known quantity of control particles comprising at least one control nucleic acid and/or at least one control protein to a biological sample, wherein the control particles comprise a non-human animal enveloped virus, wherein the non-human animal enveloped virus is selected from mouse hepatitis virus (MHV), bovine diarrhea virus (BVDV) and transmissible gastroenteritis coronavirus (TGEV);
- (b) isolating microvesicles and the control particles from the biological sample;
- (c) extracting nucleic acids and/or protein from the microvesicles and the control particles;
- (d) assaying the amount of the at least one control nucleic acid and/or the at least one control protein recovered, thereby determining the amount of control particles recovered;
- (e) determining that the amount of control particles calculated in step (d) is within a predetermined range of values;
- and
- (f) subjecting the nucleic acids and/or protein from step (c) to further analysis if the amount of control particle calculated in step (d) is within the predetermine range of values;
- wherein the isolating of the microvesicles and control particles from the biological sample comprises contacting the biological sample with a capture surface under conditions sufficient to retain cell-free DNA, microvesicles, and control particles on the capture surface, and
- wherein the capture surface comprises one or more beads that are positively charged, one or more beads that are an anion exchanger functionalized with quaternary ammonium (R—CH2—N+(CH3)3), or one or more beads that are positively charged and functionalized with quaternary ammonium.
6 more elements
A method comprising:
- (a) isolating microvesicles from a biological sample;
- (b) adding a known quantity of control particles comprising at least one control nucleic acid and/or at least one control protein to the isolated microvesicles, wherein the control particles comprise a non-human animal enveloped virus, wherein the non-human animal enveloped virus is selected from mouse hepatitis virus (MHV), bovine diarrhea virus (BVDV) and transmissible gastroenteritis coronavirus (TGEV);
- (c) extracting nucleic acids and/or protein from the isolated microvesicles and the control particles;
- (d) assaying the amount of the at least one control nucleic acid and/or the at least one control protein recovered, thereby determining the amount of control particles recovered;
- (e) determining that the amount of control particles calculated in step (d) is within a predetermined range of values;
- and
- (f) subjecting the nucleic acids and/or protein from step (c) to further analysis if the amount of control particle calculated in step (d) is within the predetermine range of values;
- wherein the isolating of the microvesicles and control particles from the biological sample comprises contacting the biological sample with a capture surface under conditions sufficient to retain cell-free DNA, microvesicles, and control particles on the capture surface, and
- wherein the capture surface comprises one or more beads that are positively charged, one or more beads that are an anion exchanger functionalized with quaternary ammonium (R—CH2—N+(CH3)3), or one or more beads that are positively charged and functionalized with quaternary ammonium.
6 more elements
Dependencies are inferred from the claim text.