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Molecularly imprinted polymers for detecting microorganisms

A technology of molecular imprinting and polymers, applied in biological testing, analytical materials, measuring devices, etc., can solve problems that have not been used to detect or identify microorganisms

Inactive Publication Date: 2011-06-22
COLUMBIA BIOSYST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although molecularly imprinted methods have shown limited success in selectively binding macromolecules, these methods have not been used to detect or identify microorganisms

Method used

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  • Molecularly imprinted polymers for detecting microorganisms
  • Molecularly imprinted polymers for detecting microorganisms
  • Molecularly imprinted polymers for detecting microorganisms

Examples

Experimental program
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Effect test

Embodiment 1

[0113] as in figure 1As demonstrated in , MIPs of the invention described herein can be made by generating a template for a portion of the target molecule and polymerizing functional monomers in the presence of this template. Functional monomers are able to bind to active sites on template molecules and then polymerize in the presence of excess cross-linking agents. While polymerization can be performed in the presence of template molecules, subsequent removal of the template molecules can leave cavities with a shape and arrangement of functional groups that are complementary to those of the template molecules. Thus, the resulting MIPs are capable of exhibiting the ability and selectivity to tightly rebind the template molecule.

Embodiment 2

[0115] Figure 2A and 2B A schematic diagram showing the detection of bacteria using the MIP of the present invention. After identification of unique macromolecules associated with bacteria, templates for portions of macromolecules comprising one or more epitopes can be generated. Functional monomers can be polymerized in the presence of template molecules such that the monomers bind to active sites on the template molecules, and monomers can be further polymerized in the presence of excess cross-linking agents. Subsequent removal of the template molecule ( Figure 2A ) is able to leave a cavity with a shape and arrangement of functional groups complementary to those of the functional groups that are part of the macromolecules unique to the microbe. The resulting imprinted polymers can thus exhibit the ability and selectivity to tightly bind portions of macromolecules associated with microorganisms and identify said microorganisms ( Figure 2B ).

Embodiment 3

[0117] Using MIP to detect methicillin-resistant Staphylococcus aureus (MRSA)

[0118] One embodiment of the invention involves the detection of MRSA using the produced MIPs that bind PBP2A. In particular, MIPs can be generated using epitope amino acid sequences corresponding to different amino acid sequences within PBP2A as template molecules. For example, the template molecule can be designed to comprise the amino acid sequence (see Table 1 below) MKKIKIVPLILIVVVVGFGIYFYAS (SEQ ID NO: 1); KKIKIVPL (SEQ ID NO: 2); KIKIVPLI (SEQ ID NO: 3); QNWVQDDTF (SEQ ID NO : 4); KEYKGYKDDAVIGK (SEQ ID NO: 5); EYKGYKDD (SEQ ID NO: 6); YKGYKDDA (SEQ ID NO: 7); DKKEPLLNKFQITTS (SEQ ID NO: 8); KEPLLNKF (SEQ ID NO: 9); (SEQ ID NO: 10); PLLNKFQI (SEQ ID NO: 11); GYNVTRYEVVN (SEQ ID NO: 12); GVGEDIPSDYPFYNAQILD (SEQ ID NO: 13); DYPFYNAQ (SEQ ID NO: 14) and / or fragments thereof, They uniquely provide surface imprints with specificity for PBP2A. The MIP generated by using these amino acid sequen...

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Abstract

The invention described herein provides molecularly imprinted polymers (MIPs) that are capable of binding to a microorganism, and methods for detecting and / or identifying microorganisms utilizing Molecularly Imprinted Polymers (MIPs). The microorganisms of the invention include prokaryotes, eukaryotes, virus and prions. The methods of the invention comprise detecting all or part, including epitopes, of macromolecules associated with the microorganisms. The macromolecules of the invention include polysaccharides, proteins, glycoproteins, peptidoglycans, lipoproteins, peptides, polypeptides, and polynucleotides, associated with said microorganisms. The invention also provides for methods of diagnosing a subject infected with the microorganisms utilizing MIPs, in addition to diagnostic kits.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Application No. 61 / 076,353, filed June 27, 2008, and U.S. Provisional Application No. 61 / 169,450, filed April 15, 2009, the disclosures of which are both adopted by References are specifically incorporated herein in their entirety. field of invention [0003] The invention described herein relates generally to molecularly imprinted polymers (MIPs) capable of binding microorganisms, including methods and kits for detecting, identifying and / or quantifying microorganisms using MIPs. Background of the invention [0004] MIPs are engineered cross-linked polymers that exhibit high affinity and selectivity for a single compound or a family of related compounds. MIPs are capable of binding analytes even when the analytes are present in complex matrices (eg, plasma, urine, muscle tissue, food matrices, environmental samples, process solutions, etc.). An important strength o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K17/08G01N33/68
CPCG01N33/56938C07K17/08
Inventor 罗伯特·L·琼斯
Owner COLUMBIA BIOSYST
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