University of Kansas

Kansas State University Research Foundation

University of California

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Mei He

Yong Zeng

Amy E Herr

Steven K Libutti

Peng Zhang

Dohyun Kim

Andrew Godwin

Chenlu Hou

Zheng Zhao

Samuel Tia

Methods for producing engineered exosomes and other vesicle-like biological targets, including allowing a target vesicle-like structure to react and bind with immunomagnetic particles; capturing the immunomagnetic particle/vesicle complex by applying a magnetic field; further engineering the captured vesicles by surface modifying with additional active moieties or internally loading with active agents; and releasing the engineered vesicle-like structures, such as by photolytically cleaving a linkage between the particle and engineered vesicle-like structures, thereby releasing intact vesicle-like structures which can act as delivery vehicles for therapeutic treatments.

Methods for generating therapeutic delivery platforms

A graphene-based sandwich immunoassay for detecting whether a target biological substance is present in a sample, generally comprising contacting said sample with a plurality of particles coated with graphene nanosheets, each particle having at least one targeting receptor, such that the target biological substance, if present, associates with the targeting receptor, and detecting the presence of the target biological substance in the sample by subsequently contacting the sample with a detection antibody, wherein the detection antibody is capable of targeting and binding with the target biological substance if bound to the targeting receptor to yield a detectable complex. The targeting receptor can be an antibody or fragment thereof. The target biological substance can be an exosome.

Graphene oxide-based nanolab and methods of detecting of exosomes

A capture device for capturing a biological substance can include: a substrate; a graphene-oxide layer on the substrate; at least one polydopamine polymer coupled with the graphene-oxide; and at least one targeting receptor coupled to the polydopamine(s), wherein the targeting receptor is capable of targeting/binding with a target biological substance. The graphene-oxide may be covalently coupled with the substrate and polydopamine, and the polydopamine may be covalently coupled with the targeting receptor. The targeting receptor can be an antibody or fragment thereof. The target biological substance can be an exosome. The substrate can be a particle (e.g., magnetic, such as magnetically responsive) or a surface in a microfluidic channel. The surface can be a top surface of a post, the post having a Y-shaped cross-sectional profile. In one aspect, the substrate is a particle. The capture device can include the target biological substance bound to the targeting receptor.

A microfluidic exosome profiling platform integrating exosome isolation and targeted proteomic analysis is disclosed. This platform is capable of quantitative exosomal biomarker profiling directly from 30 &#x3bc;L plasma samples within approximately 100 minutes with markedly enhanced sensitivity and specificity. Identification of distinct subpopulation of patient-derived exosomes is demonstrated by probing surface proteins and multiparameter analyses of intravesicular biomarkers in the selected subpopulation. The expression of IGF-1R and its phosphorylation level in non-small cell lung cancer (NSCLC) patient plasma is assessed, as a non-invasive alternative to the conventional biopsy and immunohistochemistry. The microfluidic chip, which may be fabricated of a glass substrate and a layer of poly(dimethylsiloxane), can include a first capture chamber, a second capture chamber, a serpentine microchannel, a first microchannel, a second microchannel, a sample inlet, a buffer inlet, a bead inlet, at least a first connector channel, and a reagent inlet.

Non-invasive monitoring cancer using integrated microfluidic profiling of circulating microvesicles

A microfluidic exosome profiling platform integrating exosome isolation and targeted proteomic analysis is disclosed. This platform is capable of quantitative exosomal biomarker profiling directly from plasma samples with markedly enhanced sensitivity and specificity. Identification of distinct subpopulation of patient-derived exosomes is demonstrated by probing surface proteins and multiparameter analyzes of intravesicular biomarkers in the selected subpopulation. The expression of IGF-1R and its phosphorylation level in non-small cell lung cancer (NSCLC) patient plasma is assessed as a non-invasive alternative to the conventional biopsy and immunohistochemistry. Detection of ovarian cancer also is assessed. The microfluidic chip, which may be fabricated of a glass substrate and a layer of poly(dimethylsiloxane), includes a serpentine microchannel to mix a fluid and a microchamber for the collection and detection of exosomes.

Multi-directional microfluidic devices and methods for using the same are provided. Aspects of the present disclosure include microfluidic devices that include a chamber having a separation medium, a first binding medium, and a second binding medium. In addition, the devices include a flow field element configured to subject the chamber to two or more directionally distinct flow fields. Methods of using the devices, as well as systems and kits that include the devices are also provided. The devices, systems and methods find use in a variety of different applications, including diagnostic, research and validation assays.

Microfluidic devices and methods for assaying a fluid sample using the same

A gene profiling signature for diagnosis and prognosis of cancer patients is disclosed herein. In one embodiment, the gene signature includes 32 or 79 cancer survival factor-associated genes. Thus, provided herein is a method of determining the prognosis of a subject with a tumor by detecting expression of five of more cancer survival factor-associated genes in a tumor sample and comparing expression of the five or more cancer survival factor-associated genes in the tumor sample to a control. In some examples, an increase in expression of ABCF1, CORO1C, DPP3, PREB, UBE3A, and PTDSS1 in a tumor sample compared to a control sample indicates poor prognosis. Further provided are arrays including probes or antibodies specific for a plurality of cancer survival factor-associated genes or proteins.

Molecular-based method of cancer diagnosis and prognosis

Multi-directional microfluidic devices and methods for using the same are provided. Aspects of the invention include microfluidic devices that are configured to subject a sample to two or more directionally distinct flow fields, and include a separation medium and a binding medium, where the binding medium is in fluid communication with the separation medium. Also provided are methods of using the devices as well as systems and kits that include the devices. The devices, systems and methods find use in a variety of different applications, including diagnostic and validation assays.

Multi-directional microfluidic devices and methods

Microfluidic devices having a protein renaturation component and methods for using the same are provided. Aspects of the present disclosure include microfluidic devices that include a separation medium with a first flow path and a protein renaturation component in fluid communication with the separation medium and having a second flow path. Also provided are methods of using the devices as well as systems and kits that include the devices. The devices, systems and methods find use in a variety of different applications, including diagnostic and validation assays.

Protein renaturation microfluidic devices and methods of making and using the same

A gene profiling signature for diagnosis and prognosis of cancer patients is disclosed herein. In one embodiment, the gene signature includes 32 or 79 cancer survival factor-associated genes. Thus, provided herein is a method of determining the prognosis of a subject with a tumor by detecting expression of five of more cancer survival factor-associated genes in a tumor sample and comparing expression of the five or more cancer survival factor-associated genes in the tumor sample to a control. In some examples, an increase in expression of ABCF1, CORO1C, DPP3, PREB, UBE3A, and PTDSS1 in a tumor sample compared to a control sample indicates poor prognosis. Also provided is a method of treating a patient diagnosed with cancer by administering a therapeutically effective amount of an agent that alters expression or activity of one or more of the disclosed cancer survival factor-associated genes. Further provided are arrays including probes or antibodies specific for a plurality of cancer survival factor-associated genes or proteins.

Growing community of inventors

Olathe, KS, United States of America

Mei He