Affinity binding-based system for detecting particulates in a fluid

• US 6,953,550 B2
• Filed: 11/30/2003
• Issued: 10/11/2005
• Est. Priority Date: 12/20/1996
• Status: Expired due to Fees

• Current Assignee: Tecan Trading AG (Tecan Group AG)
• Original Assignee: Tecan Trading AG (Tecan Group AG)
• Inventors: Sheppard, Norman F. Jr., Carvalho, Bruce L., Mian, Alec, Kellogg, Gregory, Kieffer-Higgins, Stephen G.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): Do, Pensee T.
• Application Number:
  US10/726,337
• Publication Number:
  US 20040142494A1
• Time in Patent Office: 681 Days
• Field of Search: 432/63, 432/64, 432/68.1, 432/50, 432/72, 432/101, 432/102, 432/67, 436/45, 436/63, 436/177, 436/180, 210/787, 210/380.1, 210/374, 210/780, 210/109, 210/781, 435/7.1, 435/7.2, 435/7.21
• US Class Current: 422/63
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 35/71725 using centrifugal forces
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/024 Storing results with means integrated into the container
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0806 Standardised forms, e.g. compact disc [CD] format
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0661 shape memory polymer valves
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 3/5027 : by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 7/00 : Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• CPC Class Codes: C07K 14/79 : Transferrins, e.g. lactoferrins, ovotransferrins
• CPC Class Codes: C12N 9/2462 : Lysozyme (3.2.1.17)
• CPC Class Codes: G01N 15/14 : Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 21/07 : Centrifugal type cuvettes G01N21/09 takes precedence
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/569 : for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 33/68 : involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 35/00069 : whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk
• CPC Class Codes: Y10S 366/03 : Micromixers: variable geometry from the pathway influences mixing/agitation of non-laminar fluid flow
• CPC Class Codes: Y10T 436/111666 : Utilizing a centrifuge or compartmented rotor
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

This invention provides methods and apparatus for detecting and quantifying particulate matter suspended in a fluid. Specifically, the invention provides an integrated, affinity-binding based, analytical system comprising a platform for performing an affinity-binding based assay for specifically binding particulates including microbial cells, and a detection means for detecting the particulates specifically bound to a defined surface or chamber comprising the platform. Methods for using the analytical systems of the invention are also provided.

Claims

1. An apparatus for detecting a particulate in a fluid, the apparatus comprising

a platform comprising a substrate having a surface, wherein said surface defines a detection chamber comprising an area that is coated with a specific binding reagent that specifically binds to the particulate to be detected;

a fluid sample input means in fluid communication with the detection chamber that is coated with the specific binding reagent;

a wash buffer reservoir containing a wash buffer in fluid communication with the detection chamber that is coated with the specific binding reagent;

a fluid waste receptacle in fluid communication with the detection chamber that is coated with the specific binding reagent;

wherein an amount of a fluid sample comprising a particulate is moved from the fluid sample input means to the detection chamber and incubated thereon for a time sufficient to result in specific binding between the particulate in the fluid sample and the specific binding reagent in the detection chamber; and

wherein the fluid sample is replaced with the wash buffer and displaced into the fluid waste receptacle; and

wherein the wash buffer is further displaced into the fluid waste receptacle; and

wherein a particulate specifically bound to the detection chamber is specifically bound to the specific binding reagent and is detected thereupon, further comprising a device, wherein the device has a surface or cavity that accommodates the platform, and a light source positioned to illuminate the platform and wherein the detection chamber further comprises alternating transparent and reflective regions.

2. An apparatus according to claim 1, wherein the alternating transparent and reflective regions define a pattern in the detection chamber.

3. An apparatus according to claim 1 wherein the specific binding reagent is present on the surface of the platform on a transparent region thereof.

4. An apparatus for detecting a particulate in a fluid, the apparatus comprising

a platform comprising a substrate having a surface, wherein said surface defines a detection chamber comprising an area that is coated with a specific binding reagent that specifically binds to the particulate to be detected;

a fluid sample input means in fluid communication with the detection chamber that is coated with the specific binding reagent;

a wash buffer reservoir containing a wash buffer in fluid communication with the detection chamber that is coated with the specific binding reagent;

a fluid waste receptacle in fluid communication with the detection chamber that is coated with the specific binding reagent;

wherein an amount of a fluid sample comprising a particulate is moved from the fluid sample input means to the detection chamber and incubated thereon for a time sufficient to result in specific binding between the articulate in the fluid sample and the specific binding reagent in the detection chamber;

wherein the fluid sample is replaced with the wash buffer and displaced into the fluid waste receptacle; and

wherein the wash buffer is further displaced into the fluid waste receptacle; and

wherein a particulate specifically bound to the detection chamber is specifically bound to the specific binding reagent and is detected thereupon, wherein the detection chamber that is coated with a specific binding reagent is further treated with a blocking agent that prevents non-specific binding to the surface of the platform, and wherein the specific binding reagent is present in the detection chamber on transparent regions thereof, and the blocking reagent is present in the detection chamber on transparent and reflective regions thereof.