Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Microfluidic System and Method for Real-Time Measurement of Antibody-Antigen Binding and Analyte Detection

a microfluidic system and antibody technology, applied in fluorescence/phosphorescence, laboratory glassware, instruments, etc., can solve the problems of inability to monitor the continuous analyte immunoassay and pharmacokinetic characterization of biomolecules in real-time, laborious and time-intensive procedures, and methods that are impractical for real-time monitoring

Inactive Publication Date: 2017-03-16
THE GENERAL HOSPITAL CORP +1
View PDF3 Cites 30 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides systems and methods for optical detection and quantification of analytes in samples using microfluidic devices. The devices include a microfluidic chip with a detector region that can detect the analyte and measure the binding affinity of an antibody to the analyte. The devices also include a light source, a light sensor, and a processor for analyzing the light sensor data. The invention allows for continuous flow optical detection and quantification of analytes in samples, with high sensitivity and accuracy. The invention also includes a system for delivering samples to the microfluidic device and a method of determining the concentration of analyte in a liquid sample.

Problems solved by technology

A major challenge in the detection of soluble molecules such as cytokines, protein antigens and antibodies is the ability to monitor time-varying or dynamic concentrations in real-time.
Currently there are no available online monitoring approaches for continuous analyte immunoassays and pharmacokinetic characterization of biomolecules in real-time.
These diagnostic methods are performed on samples obtained at pre-defined times and are therefore laborious and time-intensive procedures.
Additionally, these methods are impractical for real-time monitoring since they cannot be performed rapidly enough to assess dynamic fluctuation of analyte concentration in vivo.
This limits their utility in clinical settings where it is of critical importance to generate real-time profile of analytes such as cytokines or administered drugs in vivo (Crowther, 2001; Mannerstedt et al., 2010; Mao et al., 2009; Wild, 2001).
Nevertheless, most of these methods still require incubation and are unable to measure the dynamic changes in the analyte concentration in real time (Hu and Gao, 2007; Singhal et al., 2010).
Although diffusion distances in microchannels are significantly reduced in comparison to conventional microtiter well plate formats, analytes are still transport-limited in micro channels at low sample concentrations (Parsa et al., 2008).

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
  • Microfluidic System and Method for Real-Time Measurement of Antibody-Antigen Binding and Analyte Detection
  • Microfluidic System and Method for Real-Time Measurement of Antibody-Antigen Binding and Analyte Detection
  • Microfluidic System and Method for Real-Time Measurement of Antibody-Antigen Binding and Analyte Detection

Examples

Experimental program
Comparison scheme
Effect test

example 1

Materials and Methods

[0138]Microfluidic Device Fabrication.

[0139]The polydimethylsiloxane (PDMS) microfluidic device was fabricated using well-established soft lithography method. Negative photo resist SU-8 2100 (MicroChem, Newton, Mass.) was spin-coated on Silicon wafers to a thickness of 150 μm, and patterned by exposure to UV light through a transparency photomask (CAD / Art Services, USA). PDMS (Sylgard 184, Dow Corning, MI) was mixed with the crosslinker (Sylgard 184 curing agent) in a ratio of 10:1, poured onto the photoresist patterns, degassed thoroughly and cured for 12 hours at 65° C. Next, the PDMS was peeled off the wafer and placed in oxygen-plasma chamber in order to bond with the glass slide. The device consisted of three inlets and two mixing channels. Tygon Micro Bore PVC Tubing 0.010″ ID, 0.030″ OD, 0.010″ Wall (Small Parts Inc., FL, USA) was connected to the channels and to 1 mL syringes. Syringe pumps (Harvard Apparatus, USA) were used to maintain a flow rate of 5 ...

example 2

Flow Dynamics in a Microfluidic Device

[0185]FIGS. 1A-1C schematically illustrates the developed flow-through LOC device. The device consisted of three inlets and two mixing regions. The inlets were connected with syringe pumps that were operated individually to obtain desired flow rates for the detection of microspheres downstream. First, a solution containing the analyte molecules was introduced into inlet 1 (110) and mixed with a suspension of functionalized microspheres that were introduced via inlet 2 (120) to capture the target analyte in the first mixing channel (140) (FIGS. 1A-1C). The specific interaction that occurs between the conjugated microspheres and the target analyte in the first mixing channel leads to the analyte recognition and capture. A detection (reporter) antibody against the analyte, conjugated with a specific fluorophore, was then introduced to the flow stream via inlet 3 (150) just before the second mixing channel (170). The sandwich complex formation, comp...

example 3

Anti-TNF-α Antibody Immunoassay

[0187]To demonstrate the real-time detection capabilities of the LOC device, efforts were focused on detecting anti-TNF-α antibody. FIG. 12A describes the microsphere-based assay that was introduced into microfluidic format for anti-TNF-α detection. Avidinilated microspheres were conjugated off-chip to biotinylated human TNF-α protein via avidin-biotin bridge (Konry et al., 2009; Diamdandis et al. 1991) as described in Example 1. Next, the generated anti-TNF-α microsphere-based sensors were introduced into the microfluidic device via inlet 2 while the analyte, mouse monoclonal anti-human TNF-α antibody, was introduced via inlet 1. The interaction of the two components resulted in the capture of anti-TNF-α antibodies by microsphere-based sensors in the first mixing channel of the device. Next, the detection antibody, FITC-labeled anti-mouse IgG, was introduced into inlet 3. The fluorescent signal on the microsphere sensor, generated by the conjugation o...

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
areaaaaaaaaaaa
cross-sectional areasaaaaaaaaaa
fluidic path lengthaaaaaaaaaa
Login to View More

Abstract

Microfluidic devices for use with reagents bound to microspheres for determination of the concentration of an analyte in a liquid sample are provided. The devices include two sequential mixing channels that promote rapid binding of microsphere-bound reagents with reagents in solution and a means for detecting labeled microsphere-bound reaction products. Also provided are methods for using the devices with microsphere-bound reagents to determine the concentration of an analyte in a liquid sample and to measure the binding affinity of antibody for an antigen.

Description

BACKGROUND[0001]Rapid, sensitive and quantitative detection methods of disease markers are necessary for timely and effective diagnosis and therapy (Martinez et al., 2008). A major challenge in the detection of soluble molecules such as cytokines, protein antigens and antibodies is the ability to monitor time-varying or dynamic concentrations in real-time. Currently there are no available online monitoring approaches for continuous analyte immunoassays and pharmacokinetic characterization of biomolecules in real-time. At present, state-of-the-art analyte detection techniques for biomolecules include immunoassays such as enzyme-linked immunosorbent assays (ELISA), which are based on specific recognition of clinical antigens by the respective antibodies (Reichert, 2001). These diagnostic methods are performed on samples obtained at pre-defined times and are therefore laborious and time-intensive procedures. Additionally, these methods are impractical for real-time monitoring since the...

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
Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/543B01F5/06B01F13/00B01L3/00G01N21/64
CPCG01N33/54313B01L3/502715B01L3/502776B01L3/502761G01N33/54366G01N21/6428G01N2201/061B01F5/0647B01L2300/0654B01L2200/0605B01L2200/0647B01L2300/0883G01N2021/6439B01F13/0059B01L3/5027B01L2300/0816B01L2300/0867B01L2300/087B01L2400/0487G01N33/5304B01F25/4331B01F33/30
Inventor KONRY, TANIAYARMUSH, MARTIN L.
Owner THE GENERAL HOSPITAL CORP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com