Peptide or protein microassay method and apparatus

• US 7,332,286 B2
• Filed: 11/07/2001
• Issued: 02/19/2008
• Est. Priority Date: 02/02/2001
• Status: Expired due to Fees

• Current Assignee: University of Pennsylvania
• Original Assignee: University of Pennsylvania
• Inventors: Diamond, Scott L.
• Primary Examiner(s): LAM, ANN Y
• Application Number:
  US10/036,066
• Publication Number:
  US 20020142351A1
• Time in Patent Office: 2,295 Days
• Field of Search: 435/7.1, 435/4, 435/6, 435/287.1, 435/287.2, 435/287.3, 435/283, 422/63, 436/514, 436/517, 347/56, 347/2, 347/54, 427/466, 427/248.1, 427/457
• US Class Current: 435/7.1
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: G01N 33/552 Glass or silica

Abstract

A peptide or protein microassay method and apparatus in which a wide variety of chromogenic or fluorogenic peptide or protein substrates of interest are individually suspended or dissolved in a hydrophilic carrier, with aliquots of each substrate being deposited in an array or microarray of reaction loci, or “dots.” Each dot, therefore, provides an individual reaction vessel containing the peptide or protein of interest, to which a biological sample may be applied for assay purposes. The sample is applied to the array or microarray of dots by one of a variety of focused sample application techniques, including aerosolizing or misting of the sample, or target application of the sample, onto each dot without creating fluid channels between the dots which would cause cross-contamination.

Claims

1. An assay system, comprising:

a computer and a set of operating instructions resident in computer software of the computer for operating;

a set of reactant dot applicator pins;

an xy positioner operatively connected to the reactant dot applicator pins, wherein said reactant dot applicator pins create a microarray of liquid hydrophilic reaction dots on a planar surface, each of said reactant dots adhering to said planar surface in a non-covalent manner and having a diameter ranging from 10 microns to 100 microns and being separated by a center to center distance of 50 microns to 500 microns, and having one or more constituents therein; and

a separate device for biological sample aerosol mist generation, wherein the aerosolized biological sample mist droplets are applied simultaneously to said microarray by said separate device for sample aerosol mist generation, without forming a wetting film, for computer-enhanced assay of any reaction between the sample mist droplets and said constituents.

2. The assay system of claim 1, wherein said device for biological sample aerosol mist generation comprises one or more subcomponents and wherein said operating instructions send signals, via serial or parallel port, to start, to stop, to establish operating set points and to control said one or more subcomponents of the device, whereby each of said one or more subcomponents may have an internal or external standing controller or driver.

3. The assay system of claim 2, wherein said one or more subcomponents further comprises at least one device selected from the group consisting of multiple positive displacement microsyringe pumps, pressure nozzles, ultrasonic nozzles, ink-jet printheads, position-actuated ink-jet printheads, surface-actuated ink-jet printheads, fluid-contacting or fluid-noncontacting ultrasound transducers;

gas flow meter and controller; and

exhaust and filtration fan.

4. The assay system of claim 1, wherein said pins hold 1.0 microliters to 1000 microliters of biological sample.

5. The assay system of claim 3, wherein said microsyringe pumps deliver samples to said microarray at a constant flow rate.

6. The assay system of claim 1, wherein said device for aerosol generation is an ultrasonic nebulizer.

7. The assay system of claim 1, wherein said reaction dots comprise a carrier selected from the group consisting of dextran, pluronic acid, carbohydrates of the pentose, ribose or hexose families, polysaccharides, polyethylene glycol polymer, 1,2-ethanediol, 2,3-butanediol, and 1,2,3-propanetriol (glycerol).

8. The assay system of claim 7, wherein said reaction dots further comprise enzyme reaction components selected from the group consisting of cofactors, inhibitors, antibodies, activators, and buffer elements.

9. The assay system of claim 7, wherein said reaction dots comprise a biological molecule or fraction selected from the group consisting of proteins, peptides, nucleic acids, enzymes, antibodies, lipids, cell lysates, and vesicles.

10. The assay system of claim 7, wherein said reaction dots further comprise fluorogenic substrates, chromogenic substrates, or other reporter substrates.