Reagents and methods for the detection and quantification of thyroxine in fluid samples

• US 5,691,456 A
• Filed: 05/18/1995
• Issued: 11/25/1997
• Est. Priority Date: 03/30/1992
• Status: Expired due to Term

• Current Assignee: Abbott Laboratories
• Original Assignee: Abbott Laboratories
• Inventors: Adamczyk, Maciej, Mattingly, Phillip G., Tyner, Joan D., Johnson, Donald D., Perkowitz, Mary M., Clarisse, Diana E.
• Primary Examiner(s): CEPERLEY, MARY
• Application Number:
  US08/444,128
• Time in Patent Office: 922 Days
• Field of Search: 435/188, 530/807, 530/405
• US Class Current: 530/405
• CPC Class Codes: C07D 493/10 Spiro-condensed systems
• CPC Class Codes: C07F 9/6561 containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C09B 11/08 Phthaleins; Phenolphthaleins; Fluorescein
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/78 Thyroid gland hormones , e.g. T3, T4, TBH, TBG or their receptors
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 530/807 Hapten conjugated with peptide or protein

Abstract

Immunoassay methods and reagents for the specific quantification of thyroxine in a test sample are disclosed employing antibodies prepared with thyroxine derivatives of the formula: ##STR1## wherein P is an immunogenic carrier material and X is a linking moiety. The present invention also describes the synthesis of unique labelled reagent of the formula: ##STR2## wherein Q is a detectable moiety and W is a linking moiety, preferably fluorescein or a fluorescein derivative.

Claims

1. A method of making an immunogen of the formula:

##STR10## wherein BSA denotes bovine serum albumin, the degree of substitution of BSA by the thyroxine derivative is between 1% to 100%, the thyroxine is an L-isomer, and antibodies directed against the immunogen immunoreact with an L-thyroxine conjugate of the formula;

##STR11## where Q is a detectable moiety and W is a linking moiety, the method comprising the steps of;

(a) N-acetylating L-thyroxine to form α

-N-acetyl L-thyroxine;

(b) conjugating the α

N-acetyl L-thyroxine to 6-aminocaproic acid to form a L-thyroxine conjugate; and

(c) conjugating the L-thyroxine conjugate of step (b) to bovine serum albumin.