Methods for reducing the range in concentrations of analyte species in a sample

a technology of analyte and concentration range, which is applied in the field of combinatorial chemistry, protein chemistry and biochemistry, can solve the problems of inability to detect analyte species, inability to accurately measure the amount, and interference with the ability to detect less abundant analytes, so as to maintain the diversity of the population of analyte species

Inactive Publication Date: 2005-11-03
BIO RAD LAB INC +1
View PDF10 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0010] This invention provides a method to compress the range of concentrations between different analyte species in a complex sample while substantially maintaining the diversity of the population of analyte species in the sample. More specif...

Problems solved by technology

When the amount of an analyte in a sample is above the dynamic range, its signal saturates the detection system and the amount cannot be measured accurately.
When the amount of an analyte in a sample is below the sensitivity range of th...

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
  • Methods for reducing the range in concentrations of analyte species in a sample
  • Methods for reducing the range in concentrations of analyte species in a sample
  • Methods for reducing the range in concentrations of analyte species in a sample

Examples

Experimental program
Comparison scheme
Effect test

example 1

Reduction of Range of Concentrations of Human Serum Proteins

[0172] This example illustrates how one embodiment of the invention described above may be applied to a complex biological sample, in this case human serum. In this example, a reduction in the variance of serum protein concentrations is achieved by selectively adsorbing serum proteins to hexapeptides coupled to insoluble beads. More than 1×106 possible permutations of hexapeptide are represented in the binding moiety population of the example, in the form of a combinatorial library of split, recombine and pool beads. In this format, high abundance serum analytes, such as albumin, are bound to a hexapeptide binding moiety, but only to a level equal to saturation of the particular binding moiety. In contrast, low abundance serum analytes are bound almost in their entirety, as the amount of binding moiety recognizing the low abundance analyte is not limiting. The result of this selective binding is a reduction in the range of...

example 2

Incubation of Library with Unfractionated, Undiluted Human Pooled Plasma

[0177] To aid analysis of complex samples, this method is useful to decrease the concentration differential. Human plasma is one of the most complex and difficult to analyze materials: proteins are present in concentration range greater than 1010 (Anderson and Anderson); decreasing this range will aid in the analysis of trace proteins. Under the conditions of this method, incubation of plasma with the ligand library will increase the number of proteins that can be detected and subsequently analyzed as compared with analysis of the unprocessed starting material.

[0178] A. Sample Preparation

[0179] Frozen, pooled, human platelet-poor plasma (PPP) was thawed at 37° C. and filtered through 0.8 and 0.45 μm filters. Four replicates of approximately 1 ml of a library of hexamer peptide ligands on Toyopearl 650 M amino resin (65 μm average diameter, ˜2×106 beads / ml; Tosoh Biosciences, Montgomeryville, Pa.) with EACA-Al...

example 3

Reduction of Concentration Variance After Removal of IgG.

[0185] In many proteomic applications, one of the first steps of sample preparation is removal of albumin and IgGs, as these high abundance proteins mask the detection of lower abundance species. Removal of these proteins, however, also often removes trace species associated with them, and also involves loss of sample. It would be advantageous to have a method of sample preparation that does not require IgG depletion before analysis. This example demonstrates that removal of IgGs is not required to visualize protein species that are not detected in intact plasma. The pattern of proteins detected in LDS-PAGE is compared in plasma that has and has not been depleted of IgGs.

[0186] A. Sample Preparation

[0187] Frozen, pooled, human platelet-poor plasma (PPP) was thawed at 37° C. and filtered through 0.8 and 0.45 μm filters. IgG was removed from the plasma as follows: 5 ml Protein G Sepharose Fast-flow resin (Amersham, T&S) was p...

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
Diameteraaaaaaaaaa
Massaaaaaaaaaa
Poweraaaaaaaaaa
Login to view more

Abstract

The present invention relates to the fields of molecular biology, combinatorial chemistry and biochemistry. Particularly, the present invention describes methods and kits for dynamically reducing the variance between analyte taken from complex mixtures.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS [0001] This is a continuation-in-part application claiming the benefit of provisional application Ser. No. 60 / 559,108, filed Apr. 2, 2004, provisional application Ser. No. 60 / 582,650, filed Jun. 23, 2004, provisional application Ser. No. 60 / 587,585, filed Jul. 12, 2004, provisional application Ser. No. 60 / 643,483, filed Jan. 12, 2005, and European patent application Ser. No. 04290775.8, filed Mar. 23, 2004, the disclosures of which are incorporated in their entireties herein by reference.FIELD OF THE INVENTION [0002] The present invention relates to the fields of combinatorial chemistry, protein chemistry and biochemistry. BACKGROUND OF THE INVENTION [0003] Proteomics seeks to generate an identity profile of the entire proteome of an organism and, through analysis of this information, to identify potential diagnostic and therapeutic entities. Current technologies for resolving protein mixtures include two-dimensional gel electrophoresis and m...

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
IPC IPC(8): C12Q1/68G01N33/53G01N33/543G01N33/68
CPCC40B30/04G01N33/6803G01N33/54306G01N33/543G01N33/6845
Inventor BOSCHETTI, EGISTOHAMMOND, DAVID
Owner BIO RAD LAB INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap