# US 8,999,646 B2 · Quality control bioassays for nutriceutical and medicinal products

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- Number: US 8,999,646 B2
- Document ID: US8999646B2
- Status: Granted
- Kind: B2
- Application number: 13/264,086
- Filed: 2010-04-16
- Published: 2012-05-31
- Granted: 2015-04-07
- Earliest claimed: 2009-04-17
- Assignees: President and Fellows of Harvard College
- Applicants: Jose A. Halperin; Huseyin Aktas
- Inventors: Jose A. Halperin; Huseyin Aktas
- Web page: https://patentgrep.com/patent/US8999646B2

## CPC

- G01N 33/53: Immunoassay; Biospecific binding assay; Materials therefor
- C12Q 1/6897: involving reporter genes operably linked to promoters
- G01N 33/68: involving proteins, peptides or amino acids {(involving lipoproteins G01N33/92)}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

Bioassays for detecting the ability of one sample of a food substance, nutritional supplement, therapeutic agent and/or disease preventive agent relative to that of a second sample of such a substance, supplement and/or agent to inhibit, upregulate or otherwise modulate translation initiation, and thereby demonstrate a disease curative and/or preventive effect in a human and/or animal that consumes a such substance, supplement and/or agent or to whom a such substance, supplement and/or agent is administered are provided.

## Claims

### Claim 1

An in vitro method for identifying whether a fish oil sample contains n-3 polyunsaturated fatty acid (PUFA) activity comprising the steps of:
providing a first test system including a first mRNA sequence encoded by a first biomarker gene having a coding region for a first reporter protein operably linked to a first biomarker promoter;
providing a second test system including a second mRNA sequence encoded by a second biomarker gene having a coding region for a second reporter protein operably linked to a second biomarker promoter;
contacting said second test system with the fish oil sample; and
detecting transcription levels of said first and said second mRNA sequences, and
identifying the fish oil sample as containing PUFA activity if the sample mediates transcriptional upregulation of the second biomarker gene or wherein the transcription level of said second mRNA sequence is greater than the transcription level of said first mRNA sequence, and wherein said first and said second biomarker genes are members selected from the group consisting of nucleic acid sequences that encode pro-apoptotic C/EBP homologous protein (CHOP), ER chaperone binding protein (BiP), Activating Transcription Factor 4 (ATF-4), and X-box binding protein 1 (Xbp-1).

### Claim 2 (depends on 1)

The method of claim 1, wherein said first and second test systems are selected from the group consisting of a cell and a cell free assay.

### Claim 3 (depends on 1)

The method of claim 1, wherein said first test system is contacted with a standard substance or a control substance.

### Claim 4 (depends on 1)

The method of claim 1, wherein transcription levels are determined by real time PCR.

### Claim 5 (depends on 4)

The method of claim 4, wherein the detecting step comprises real time PCR performed in vitro.

### Claim 6 (depends on 4)

The method of claim 4, wherein the detecting step comprises real time PCR performed in cells.

### Claim 7 (depends on 1)

The method of claim 1, wherein transcriptional activity is determined by detecting one or more reporter protein activities.

### Claim 8 (depends on 1)

The method of claim 1, wherein transcription of said second mRNA sequence is at least 150% of the transcription level of said first mRNA sequence.

### Claim 9 (depends on 1)

The method of claim 1, wherein the fish oil sample is selected from the group consisting of a food product sample, a nutriceutical product sample and a pharmaceutical product sample.

### Claim 10 (depends on 1)

The method of claim 1, wherein the fish oil sample contains eicosapentaenoic acid (EPA).

### Claim 11 (depends on 1)

The method of claim 1, wherein said n-3 PUFA activity is a member selected from the group consisting of eicosapentaenoic acid (EPA) activity, a-linoleic acid (ALA) activity and docosahexaenoic acid (DHA) activity.

### Claim 12 (depends on 10)

The method of claim 10 wherein the fish oil sample is from a fish selected from the group consisting of cold water fish, warm water fish, fresh water fish, salt water fish, brackish water fish, wild fish and farm-raised fish.

### Claim 13 (depends on 12)

The method of claim 12 wherein the fish is a salmon, mackerel or sardine.