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Simplified extraction methods for the rapid determination of species content of adipose tissue based on the detection of tni in immunoassays

a technology of tni and immunoassay, applied in the field of soluble protein extraction from adipose tissue, can solve the problems that the existing methods of extracting proteins from adipose tissue are often not particularly easy or quick to perform

Inactive Publication Date: 2015-08-20
FLORIDA STATE UNIV RES FOUND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a method for separating solidified fat and other solid materials from a mixture containing soluble proteins and insoluble / immiscible substances. The method involves separating the aqueous phase containing the soluble proteins from the mixture by separating the solid materials from the aqueous phase and filtering the aqueous phase through a filter to obtain a protein extract. This technique allows for efficient separation and purification of proteins from samples containing fat.

Problems solved by technology

Existing methods of extracting proteins from adipose tissue are often not particularly easy or quick to perform.

Method used

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  • Simplified extraction methods for the rapid determination of species content of adipose tissue based on the detection of tni in immunoassays
  • Simplified extraction methods for the rapid determination of species content of adipose tissue based on the detection of tni in immunoassays

Examples

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Effect test

example 1

[0069]Detection of pork fat in fat or meat mixtures with the sandwich ELISA using pork specific anti-TnI MAbs 5H9 and 8F10.

Methodology

Sample Preparation

[0070]All visible muscle or blood is trimmed from the white adipose tissue. The surface of the sample is then rinsed by clean pure water. The sample is then patted dry and then ground twice using a household meat grinder. Lean meat samples are ground the same way as the fat samples.

Extracting Pork Fat Proteins from Pork Fat Fortified in Beef Fat (or Chicken Fat) Mixture

[0071]For raw pork fat in beef fat (or chicken fat), a 10% (w / w) pork fat in beef fat (or chicken fat) is prepared by mixing 4.5 g of ground beef fat (or chicken fat) with 0.5 g of ground pork fat. The mixture is pre-heated in an oven for 30 minutes at 65° C. Ten percent (w / w) cooked pork fat in beef fat (or chicken fat) and autoclaved pork fat in beef fat (or chicken fat) are similarly prepared by mixing 4.5 g of beef fat (or chicken fat) with 0.5 g pork fat and then ...

example 2

Detection of Pork Fat in Fat or Meat Mixtures Using Lateral Flow Assay

[0086]Sample protein extraction was performed in the same manner as described above, then laboratory mixed sample extracts are prepared as follows for the determination of the detection limit using a lateral flow test. The lateral flow assay strips are prepared using published procedures. To make sure the homogeneity of the sample extracts containing low levels of porcine fat proteins, artificially adulterated samples are prepared from 10% (w / w) adulterated sample extracts diluted by the extract of the matrix material as described below.[0087]1. To 10% (w / w) pork fat in beef fat, pork fat in chicken fat, pork fat in beef meat, and pork fat in chicken meat adulterated sample extracts is added the appropriate amount of beef fat, chicken fat, beef meat and chicken meat extracts, respectively, to obtain 5% down to 0.3% (v / v) adulterated samples.[0088]2. To 200 μL of the diluted samples in (1) above is added 100 μL, of...

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Abstract

A method for extracting protein from a fat-containing sample is described comprising the steps of separating solidified fat and solid residues from a centrifuged mixture of protein, fat, other solid materials and aqueous solution of phosphate buffered saline to form an aqueous phase containing the protein. The aqueous phase is then filtered through a filter to separate a clear protein extract from the mixture.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims benefit of priority to U.S. Provisional Patent Application No. 61 / 693,839 filed Aug. 28, 2012, entitled “SIMPLIFIED EXTRACTION METHODS FOR THE RAPID DETERMINATION OF SPECIES CONTENT OF ADIPOSE TISSUE BASED ON THE DETECTION OF TNI IN IMMUNOASSAYS”, which is incorporated herein by reference in its entirety.BACKGROUND[0002]1. Field of the Invention[0003]The present invention relates to a method of soluble protein extraction from adipose tissue.[0004]2. Related Art[0005]Existing methods of extracting proteins from adipose tissue are often not particularly easy or quick to perform.SUMMARY[0006]According to a first broad aspect, the present invention provides a method comprising the following steps: (a) separating solidified fat and other solid materials from a mixture to thereby form an aqueous phase containing soluble proteins, and (b) filtering the aqueous phase through a filter to thereby separate a protein extract f...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N33/68
CPCC07K1/145G01N1/28G01N33/68
Inventor HSIEH, YUN-HWA PEGGY
Owner FLORIDA STATE UNIV RES FOUND INC