Unlock instant, AI-driven research and patent intelligence for your innovation.

Compiled Methods for Analysing and Sorting Samples

a sorting and sample technology, applied in the field of assaying, can solve the problems of data acquisition not being adequately optimized with the specific goal, sample preparation, analysis or sorting,

Inactive Publication Date: 2010-09-30
DAKOAS
View PDF100 Cites 25 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The present invention is about methods for detecting, analyzing, isolating, and manipulating entities such as cells, particles, and supra-molecular structures. The method involves preparing a sample by contacting it with marker molecules specific for the entities, labelling molecules specific for the entities, and a detection molecule comprising a marker molecule and a labelling molec assaying the sample for the presence of the entities. The method can also include data processing and sample manipulations such as cultivation and expansion of isolated cells. The patent is particularly interesting because it discusses the use of MHC molecules and MHC-multimers as marker molecules."

Problems solved by technology

However, often the process of sampling, sample preparation, analysis or sorting, and data acquisition is not adequately optimized with the specific goal in mind.
As an example, flow cytometry analyses often include reagents that have not been properly design to achieve the best possible result: The individual reagents and their binding affinity or target abundance is not seriously considered and corrected for, and likewise, the fluorochromes chosen are not well enough separated as regards wavelength of maximum emission, and therefore, the spectra of the fluorochormes significantly overlap, blurring the data further.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Compiled Methods for Analysing and Sorting Samples
  • Compiled Methods for Analysing and Sorting Samples
  • Compiled Methods for Analysing and Sorting Samples

Examples

Experimental program
Comparison scheme
Effect test

example 1

“Matrix Protocol for MHC Class I Specific T-cell Quantization Performed on Human Blood”

[1455]In this example the following sub-processes are used:[1456]Sampling: Blood retrieval from person using a vacutainer, containing an anticoagulation agent.[1457]Sample preparation: Fructose / Trehalose (matrix) and fluorescent markers were present in the assay container in a Matrix. No-lyse protocol was followed, i.e. the red blood cells were not lysed.[1458]Assaying: The analysis involves the use of the following marker molecules: anti-CD45 antibody (binds to the CD45 membrane protein of leucocytes), anti-CD3 antibody (binds to CD3 protein on surface of T cells), anti-CD8 antibody (binds to CD8 receptor on surface of cytotoxic T cells and NK cells), and MHCCMVDex, binds to HLD protein on the surface on antigen specific cytotoxic T cells. CytoCOunt™ beads, for quantitation of positive cell / volume sample.[1459]Acquisition: Data was acquired using the Summit™ software of the CyAn ADP™ flow cytomet...

example 2

“A Lyse and Wash (LW) Protocol has been Employed for Chemical Contra Selection of the RBC”

[1469]In this example the following sub-processes are used:[1470]Sampling: Primary Vacutainer®, containing EDTA as anticoagulant Sample[1471]Preparation: Lyse and wash, with light fixation, using Utilyse™[1472]Assaying: Markers was identical to example 1, but here using Scatter parameters as primary gate criteria.[1473]Acquisition: Data was acquired using the Summit™ software of the CyAn ADP™ flow cytometer[1474]Sorting: ND[1475]Interpretation of data: Data were analyzed using the Summit™ software, Quantitation of the concentration of antigen specific Cytotoxic T Cells[1476]Manipulation of entities: ND

[1477]The sample, 100 μl peripheral human blood was added 100 μl UtiLyse™ solution A (Dako Cat. No. S3350), for a light sample fixation. After 10 min. incubation 2 ml UtiLyse™ solution B, the lysing reagents containing ammonium, were added and the sample were incubated 10 min at room temperature. ...

example 3

“Physical Selected Mononucleated Cells and Subsequent Analyzed Using Flowcytometre”

[1479]In this example the following sub-processes are used:[1480]Sampling: Peripheral blood form a human being, in a open blood glass.[1481]Sample preparation: Mononucleated cell fractioning using ficoll density centrifugation.[1482]Assaying: The analysis involves the use of the following marker molecules: anti-CD3 antibody (binds to the CD3 T cell receptor), anti-CD8 antibody (binds to CD8 receptor on surface of cytotoxic T cells and NK cells), and MHCCMVDex, binds to HLD protein on the surface on antigen specific cytotoxic T cells.[1483]CytoCount™ was added in the sample for enumeration of a antigen specific T-Cell population.[1484]Acquisition: Data was acquired using the Summit™ software of the CyAn ADP™ flow cytometer.[1485]Sorting: ND.[1486]Interpretation of data: Data were analyzed using the Summit™ software, and Quantitation of the concentration of antigen specific Cytotoxic T Cells.[1487]Manip...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
wavelengthaaaaaaaaaa
supra-molecular structuresaaaaaaaaaa
fluorescenceaaaaaaaaaa
Login to View More

Abstract

The present invention relates to the fields of analysis, diagnostics, prognostics, standardization, characterization and enumeration of entities such as biological cells, as well as therapeutical applications. Accordingly, the present invention in preferred aspects is directed to methods for the detection and / or analysis and / or isolation and optionally further manipulation of entities, such as cells, particles, and supra-molecular structures.

Description

[0001]This application claims priority under 35 U.S.C. §120 as a continuation of PCT Application No. PCT / DK2008 / 050168 filed Jul. 3, 2008, which claims priority to the following Danish Patent applications Nos.—PA 2007 00972, filed Jul. 3, 2007, PA 2007 00973, filed Jul. 3, 2007, PA 2007 00974, filed Jul. 3, 2007, and PA 2007 00975, filed Jul. 3, 2007, and also claims priority to the following U.S. Provisional Patent Applications Nos.—U.S. 60 / 929,581, filed Jul. 3, 2007, U.S. 60 / 929,582, filed Jul. 3, 2007, U.S. 60 / 929,583, filed Jul. 3, 2007, and U.S. 60 / 929,586, Jul. 3, 2007, the contents of each of which are hereby incorporated by reference.[0002]All patent and non-patent references cited in this application are hereby incorporated by reference in their entirety. All patent and non-patent references cited in U.S. 60 / 929,583, U.S. 60 / 929,586, U.S. 60 / 929,582, U.S. 60 / 929,581, PA 2007 00974, PA 2007 00972 and PA 2007 00975 are hereby incorporated by reference in their entirety. U.S....

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/53
CPCA61K38/00A61K47/4823C07K14/70539A61K47/48353B82Y5/00A61K47/48276A61K47/61A61K47/6425A61K47/665A61P31/00
Inventor JACOBSEN, KIVINPEDERSEN, HENRIKBRIX, LISELOTTELOHSE, JESPERSCHOLLER, JORGENJAKOBSEN, TINA
Owner DAKOAS