Combinatorial multidomain mesoporous chips and a method for fractionation, stabilization, and storage of biomolecules

• US 8,685,755 B2
• Filed: 07/20/2010
• Issued: 04/01/2014
• Est. Priority Date: 07/20/2009
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Ferrari, Mauro, Liu, Xuewu, Tasciotti, Ennio, Bouamrani, Ali, Hu, Ye
• Primary Examiner(s): Haq, Shafiqul
• Assistant Examiner(s): Hollinden, Gary E
• Application Number:
  US12/839,606
• Publication Number:
  US 20110065207A1
• Time in Patent Office: 1,351 Days
• Field of Search: None
• US Class Current: 436/527
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 33/552 Glass or silica
• CPC Class Codes: Y10T 428/2982 Particulate matter [e.g., sphere, flake, etc.]
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A new fractionation device shows desirable features for exploratory screening and biomarker discovery. The constituent MSCs may be tailored for desired pore sizes and surface properties and for the sequestration and enrichment of extremely low abundant protein and peptides in desired ranges of the mass/charge spectrum. The MSCs are effective in yielding reproducible extracts from complex biological samples as small as 10 μl in a time as short as 30 minutes. They are inexpensive to manufacture, and allow for scaled up production to attain the simultaneous processing of a large number of samples. The MSCs are multiplexed, label-free diagnostic tools with the potential of biological recognition moiety modification for enhanced specificity. The MSCs may store, protect and stabilize biological fluids, enabling the simplified and cost-effective collection and transportation of clinical samples. The MSC-based device may serve as a diagnostic tool to complement histopathology, imaging, and other conventional clinical techniques. The MSCs mediated identification of disease-specific protein signatures may help in the selection of personalized therapeutic combinations, in the real-time assessment of therapeutic efficacy and toxicity, and in the rational modulation of therapy based on the changes in the protein networks associated with the prognosis and the drug resistance of the disease.

Claims

1. A method of analyzing a biological sample, the method comprising the steps of:

a. providing the sample;

b. providing a substrate comprising a silica-based mesoporous material, wherein;

the pores of said mesoporous material have a pre-determined pore morphology;

a surface of the substrate is functionalized with a phosphate moiety to which an inorganic metal ion is immobilized; and

the inorganic metal ion is selected from the group consisting of gallium, titanium, and zirconium;

c. exposing the mesoporous material to the sample such that a fraction of the sample is retained by the mesoporous material; and

d. analyzing a fraction of the sample retained by the mesoporous material.

2. A method of detecting a marker of a physiological condition in a sample, the method comprising the steps of:

a. providing a sample from a subject affected by the physiological condition;

b. providing a substrate comprising a silica-based mesoporous material, wherein;

the pores of said mesoporous material are substantially uniform and have a pre-determined pore morphology;

a surface of the substrate is functionalized with a phosphate moiety to which an inorganic metal ion is immobilized; and

the inorganic metal ion is selected from the group consisting of gallium, titanium, and zirconium;

c. exposing the mesoporous material to the sample;

d. analyzing a fraction of the sample retained by the mesoporous material; and

e. comparing a result of the analyzing with a result of analyzing a control sample to detect the marker of the physiological condition.

3. A probe comprising a substrate that comprises a mesoporous material having a pre-determined morphology to separate a first component from a second component, wherein said probe is configured and arranged to be inserted into a mass spectrometer, wherein a surface of the substrate is functionalized with a phosphate moiety to which an inorganic metal ion is immobilized;

and

the inorganic metal ion is selected from the group consisting of gallium, titanium, and zirconium.

4. A mesoporous silica chip comprising multiple pores, wherein a surface of the mesoporous silica chip is functionalized with a phosphate moiety to which an inorganic metal ion is immobilized;

and

the inorganic metal ion is selected from the group consisting of gallium, titanium, and zirconium and wherein the pores on said chip are of different sizes and physiochemical properties.

5. The chip of claim 4, wherein said physiochemical properties are selected from the group consisting of surface area, pore size, the presence of functional groups, and the presence of an electrical charge.

6. The chip of claim 5, wherein said pores have a shape comprising cubic, hexagonal, honeycomb-like, tubular, circular, oblong, and combinations thereof.