TSH immunoassays and processes for performing TSH immunoassays in the presence of endogenous contaminants in restricted wash formats

• US 9,575,076 B2
• Filed: 10/29/2015
• Issued: 02/21/2017
• Est. Priority Date: 05/27/2011
• Status: Expired due to Fees

• Current Assignee: Abbott Point Of Care Incorporated (Abbott Laboratories)
• Original Assignee: Abbott Point Of Care Incorporated (Abbott Laboratories)
• Inventors: Wang, Dan, Collier, Gordon Bruce, Skinner, Joseph P., Ruan, Qiaoqiao, Tetin, Sergey Y.
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US14/926,375
• Publication Number:
  US 20160109465A1
• Time in Patent Office: 481 Days
• US Class Current: 1/1
• 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: C07K 16/26 against hormones ; against hormone releasing or inhibiting factors
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: G01N 2333/59 Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: G01N 33/76 Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors

Abstract

The invention relates to low wash Thyroid Stimulating Hormone (TSH) immunoassays using an ELISA sandwich assay having limited or no wash step between the antigen capture, detection antibody addition and substrate introduction steps. This invention exhibits low cross reactivity with biologically similar interfering cross reacting species, such as Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) and Chorionic Gonadotropin (CG).

Claims

1. A method of performing a single-wash step sandwich immunoassay for detecting thyroid stimulating hormone (TSH) comprising:

a) contacting a whole-blood sample with at least one signal antibody and at least one capture antibody to form a sandwich complex, wherein the at least one capture antibody is immobilized on an immunosensor;

(b) washing the whole-blood sample and uncomplexed signal antibody from the immunosensor with a wash fluid in no more than one washing step, wherein the wash fluid comprises a substrate; and

(c) detecting an electrochemical signal on the immunosensor generated from a reaction of the sandwich complex with the substrate as indicative of a level of the TSH in the whole-blood sample,

wherein a dissociation constant (Kd) of the at least one signal antibody and the at least one capture antibody for TSH is between 0.08 and 0.1 nM,

wherein a Kd of the at least one capture antibody for follicle stimulating hormone (FSH), luteinizing hormone (LH) and chorionic gonadotropin (CG) is substantially 1000 nM, 1000 nM, and 500 nM, respectively, and

wherein the at least one capture antibody is mouse monoclonal anti-TSH antibody 5409 and the at least one signal antibody is mouse monoclonal anti-TSH antibody M94206.

2. The method of claim 1, wherein a Kd of the at least one signal antibody for FSH, LH and CG is substantially 300 nM, 35 nM, and 40 nM, respectively.

3. The method of claim 1, wherein the at least one capture antibody and the at least one signal antibody, respectively, bind to at least two different epitopes on a TSH-β

subunit of TSH.

4. A method of performing a single-wash step sandwich immunoassay for detecting thyroid stimulating hormone (TSH), comprising:

a) contacting a whole-blood sample with at least one signal antibody and at least one capture antibody to form a sandwich complex, wherein the at least one capture antibody is immobilized on an immunosensor;

(b) washing the whole-blood sample and uncomplexed signal antibody from the immunosensor with a wash fluid in no more than one washing step, wherein the wash fluid comprises a substrate; and

(c) detecting an electrochemical signal on the immunosensor generated from a reaction of the sandwich complex with the substrate as indicative of a level of the TSH in the whole-blood sample,

wherein a dissociation constant (Kd) of the at least one signal antibody and the at least one capture antibody for TSH is between 0.04 and 0.1 nM,

wherein a Kd of the at least one capture antibody for follicle stimulating hormone (FSH), luteinizing hormone (LH) and chorionic gonadotropin (CG) is substantially 1000 nM, 1000 nM, and 500 nM, respectively, and

wherein the at least one capture antibody is mouse monoclonal anti-TSH antibody 5409 and the at least one signal antibody is mouse monoclonal anti-TSH antibody Ab9390.

5. The method of claim 4, wherein a Kd of the at least one signal antibody for FSH, LH and CG is substantially 70 nM, 54 nM, and 45 nM, respectively.

6. The method of claim 4, wherein the at least one capture antibody and the at least one signal antibody, respectively, bind to at least two different epitopes on a TSH-β

subunit of TSH.