United States of America as Represented by the Secretary of the Air Force

Conceptual Mindworks, Inc.

Other

Johnathan Lloyd Kiel

Jill E Parker

Eric A Holwitt

Jeevalatha Vivekananda

David N Erwin

John L Alls

John G Bruno

Mark A Sloan

David M Simmons

Homer Gifford

Pedro J Morales, Legal Representative

Ronald M Cook

Richard A Weber

Kary B Mullis

Harvey A Schwertner

Charles R Batishko

Melanie Woitaske

Kurt A Stahl

Maomian Fan

Michael (Maomian) Fan

Gerald J O'Brien

Christopher McQueen

John P Kilian

Shelly D Roper

Amanda Tijerina

Embodiments herein relate to compositions and methods for making and using aptamers, for example, DNA aptamers (DCEs) and/or RNA aptamers. In some embodiments, methods relate to making and amplifying target DCEs. In certain embodiments, methods for making capture elements or aptamers concern using a reporter moiety and signal reducing moiety prior to amplifying a target-specific capture element. In some embodiments, methods disclosed herein may be used to rapidly generate large quantities of aptamers such as DCEs directed to a particular target agent. Some embodiments relate to systems for performing automated generation of aptamers.

Methods and compositions for processes of rapid selection and production of nucleic acid aptamers

Embodiments herein report compositions, systems and methods for making and using plasmid vectors and nanotube complexes. In certain embodiments, compositions, systems and methods herein include making plasmid vectors having aptamer inserts. In some embodiments, methods disclosed herein may be used to rapidly generate large quantities of plasmid vectors having aptamer inserts directed to a particular target agent. Other aspects concern plasmid constructs associated with organic semiconductors. Yet other aspects concern complexes of nanotubes associated with dsDNA aptamers and tracking molecules.

Compositions, methods and uses for biosynthetic plasmid integrated capture elements

Methods and compositions for immediately immunizing an individual against any molecule or compound are provided. The present invention is directed to an immunity linker with at least two sites; (1) at least one first binding site that binds to an immune response component in an individual, and (2) at least one second binding site that binds specifically to a desired compound or molecule, the target. The second binding sites are preferably thiolated aptamers that have the benefit of increased stability, resistance to degradation and longer circulating half life. Methods of making and using pharmaceutical compositions including immunity linker molecules having a thiolated aptamer are also provided.

Chemically programmable immunity

The present invention concerns methods, compositions and apparatus for neutralizing bioagents, wherein bioagents comprise biowarfare agents, biohazardous agents, biological agents and/or infectious agents. The methods comprise exposing the bioagent to an organic semiconductor and exposing the bioagent and organic semiconductor to a source of energy. Although any source of energy is contemplated, in some embodiments the energy comprises visible light, ultraviolet, infrared, radiofrequency, microwave, laser radiation, pulsed corona discharge or electron beam radiation. Exemplary organic semiconductors include DAT and DALM. In certain embodiments, the organic semiconductor may be attached to one or more binding moieties, such as an antibody, antibody fragment, or nucleic acid ligand. Preferably, the binding moiety has a binding affinity for one or more bioagents to be neutralized. Other embodiments concern an apparatus comprising an organic semiconductor and an energy source. In preferred embodiments, the methods, compositions and apparatus are used for neutralizing anthrax spores.

Methods for neutralizing anthrax or anthrax spores

We describe examples using aptamers for capturing and reporting the presence of a target, such as a pathogen. Examples described here include a set of aptamers that are specific to. Other examples described here include an Aptamer-Linked Immobilized Sorbent Assay (ALISA) and dot blot assay. An example of a method provided here comprises: providing a set of DNA sequences that exhibit high binding affinity to target antigen, placing the DNA sequences in a sandwich aptamer-linked immobilized sorbent assay (ALISA), contacting the DNA sequences with a sample, and detecting whether the target is present in the sample. Some alternative implementations may include dot blots and different reporters. Quantum dot sandwich assays and quantum dot de-quenching reporters can be used.

Aptamer-based assays

A new strain ofderived from the Sterne vaccine strain ofby growth on a high-nitrate-concentration, 3-amino-L-tyrosine growth medium.

Medium and method of deriving and using a mutated bacteria strain

Methods and compositions for neutralizing anthrax and other bioagents

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A PCR-based method for the identification ofis described. The method utilizes novel primer sets; designated 2Xlg3F (SEQ ID NO 3), 2Xlg3R (SEQ ID NO 4), 2Xlg3F(SEQ ID NO 5), 2Xlg3R(SEQ ID NO 6), 4XH1aF (SEQ ID NO 7), and 4XH1aR (SEQ ID NO 8).

DNA probes for specific genes of anthrax

The present invention concerns methods of preparing nucleic acid ligands against anthrax spores, compositions comprising anthrax specific nucleic acid ligands and methods of use of such ligands for detection and/or neutralization of anthrax spores.

Methods and compositions for aptamers against anthrax

The cell line HeLa is transformed with the chromosomal insertion of the plasmid pSV,neoNR10,, ATCC No. 69617. The transformed cells, HeLaNR1, produce diazoluminomelanin (DALM) intra cellularly when provided with nitrate, luminol and 3-amino-L-tyrosine&bull;HC1 (3AT). The modified cells can be used to study mechanisms for radiofrequency and light radiation interactions with carcinoma of the cervix. The effects of drugs, hormones, and cytokines that affect the expression of nitric oxide synthase and its activity can also be studied to understand the effects of these materials on cervix cells.

Nitrate reductase-transfected HeLa cells for cancer and microwave bioeffects research

In a recognition complex system, nucleic acid ligands comprising random DNA sequences are operatively coupled to an organic semiconductor and distributed so as to form an array of recognition complexes. When an unknown chemical or biological analyte is applied to the array, the electrical and/or photochemical properties of one or more of the recognition complexes are altered upon binding of the nucleic acid ligand to the analyte. The degree to which the electrical and/or photochemical properties change is a function of the affinity of the nucleic acid ligand sequence for the analyte. The electrical and photochemical changes associated with the array, as a whole, can be used as a unique signature to identify the analyte. In certain embodiments, an iterative process of selection and amplification of nucleic acid ligands that bind to the analyte can be used to generate a new array with greater affinity and specificity for a target analyte, or to produce one or more nucleic acid ligands with high binding affinity for an analyte. The present invention also provides methods for preparing nucleic acid ligands that bind with high affinity to an analyte and using such nucleic acid ligands to neutralize the analyte.

Organic semiconductor recognition complex and system

The cell line EMT-6 are transformed with the chromosomal insertion of the plasmid pSV.sub.2 neoNR10.sub.1, ATCC No. 69617. The transformed cells, EMT-6/pSV.sub.2 neoNR10.sub.1, produce diazoluminomelanin (DALM) intracellularly when provided with nitrate, luminol and 3-amino-L-tyrosine.multidot.HCl (3AT). The modified cells can be used to study mechanisms for radiofrequency and light radiation interactions with breast tumor cells in vitro and in mice. The effects of drugs, hormones, and cytokines that affect the expression of nitric oxide synthase and its activity can also be studied to understand the effects of these materials on breast tumor cells.

Breast tumor cells for study of nonionizing radiation effects

An apparatus and method for immunomagnetic separation and concentration of target biological materials is disclosed. The immunomagnetic separation is performed by a magnetic flow cell, or filter block, as part of an automated mostly continuous immunomagnetic assay system. The magnetic flow cell has two bundles of ferromagnetic rods or pins positioned inside an internal chamber so that a fluid sample flowing through the flow cell passes through the pins. A pair of cobalt magnets flank the flow cell so that the pins concentrate and sufficiently increase the magnetic fields so that even nanometer size magnetic beads can be captured. The overall system combines a reaction subsystem for reacting coated magnetic beads with a sample, a collection subsystem for capturing magnetic beads, a rinsing subsystem for removing debris and a filtering subsystem for removing captured magnetic beads from the collection subsystem. The new magnetic flow filter is the key component for the collection and filtering subsystems.

Immunomagnetic assay system for clinical diagnosis and other purposes

There is provided a method for producing diazoluminomelanin (DALM) which comprises culturing in a medium containing nitrate, 3-amino-L-tyrosine (3-AT) and luminol under suitable metabolic conditions, a microorganism containing nitrate reductase. Also provided is a method for directly detecting microorganisms containing nitrate reductase or those into which nitrate reductase can be introduced by recombinant DNA technology, which comprises culturing the microorganism in a medium containing nitrate, 3-amino-L-tyrosine (3-AT) and luminol under suitable metabolic conditions, transferring the medium to a microtiter plate or tube coated with antibody or an antiligand to which the microorganism would specifically bind, washing the plate or tube and activating luminescence. Further, there is provided a method for producing diazomelanin (DM) which comprises culturing in a medium containing nitrate and 3-amino-L-tyrosine (3-AT) under suitable metabolic conditions, a microorganism containing nitrate reductase. Yet further, there is provided a method for directly detecting microorganisms containing nitrate reductase or those into which nitrate reductase can be introduced by recombinant DNA technology, which comprises culturing the microorganism in a medium containing nitrate and 3-amino-L-tyrosine (3-AT) under suitable metabolic conditions, transferring the medium to a microtiter plate or tube coated with antibody or an antiligand to which the microorganism would specifically bind, washing the plate or tube, adding luminol and activating luminescence.

Growing community of inventors

Universal City, TX, United States of America

Johnathan Lloyd Kiel