Claims for Patent: 5,576,219

Summary for Patent: 5,576,219

Title:	Standard solutions for determination of thyroxine-binding capacity in serum
Abstract:	The present invention provides a process for the determination of the thyroxine-binding capacity in serum, wherein, for the calibration, there are used at least two different standard solutions which contain substantially the same amount of thyroxine-binding globulin but a differing content of thyroxine. The present invention also provides a standard solution for the determination of the thyroxine-binding capacity containing thyroxine-binding globulin dissolved in a buffer system.
Inventor(s):	Bienhaus; Gerhard (Worthesee, DE), Jering; Helmut (Tutzing, DE)
Assignee:	Boehringer Mannheim GmbH (Mannheim-Waldhof, DE)
Application Number:	08/190,705
Patent Claims:	1. In a reagent kit for the determination of thyroxine binding capacity in serum wherein the kit contains reagents effective for the determination of thyroxine binding protein comprising components selected from the group consisting of antibodies against thyroxine, labeled antibodies against thyroxine, thyroxine, labeled thyroxine, effective buffers, effective standards, and detection reagents the improvement consisting of two separate serum-free standard solutions, the first standard solution containing 10 to 60 .mu.g/ml thyroxine-binding globulin and 100-500 ng/ml thyroxine dissolved in a buffer system having a pH range of 6 to 8 together with 40-80 mg/ml albumin and the second standard solution containing 10 to 60 .mu.g/ml thyroxine-binding globulin dissolved in a buffer system having a pH range of 6 to 8 together with 40-80 mg/ml albumin. 2. In a process for the determination of thyroxine-binding capacity in serum wherein a characteristic related to the capacity is determined by effective methods of determination, the improvement consisting essentially of calibrating said effective determination by means of two different serum-free standard solutions wherein a first standard solution contains a buffer system having a pH range of 6-8, 40-80 mg/ml albumin, 10 to 60 .mu.g/ml thyroxine-binding globulin thereby corresponding to the highest thyroxine-binding capacity of human serum and a second standard solution contains a buffer system having a pH rang of 6-8, 40-80 mg/ml albumin, 10 to 60 .mu.g/ml thyroxine-binding globulin and 100-500 ng/ml thyroxine thereby corresponding to the lowest thyroxine-binding capacity of human serum and adding each standard to a separate container having T4 antibody, further adding a predetermined amount of a reagent containing T4 and enzyme-conjugated T4 to each container, incubating the containers to permit competitive binding of the T4 and enzyme-conjugated T4 to thyroxine-binding protein and the anti-T4, removing unbound immunoreactants, adding substrate for the enzyme, determining, the amount of enzyme bound by measuring the colored product formed to produce a calibration curve from the values as determined above for the two standards, and determining the amount of thyroxine-binding capacity by the effective method for determination with reference to the above calibration curve. 3. The process of claim 2 wherein said albumin is human serum albumin, bovine serum albumin or horse serum albumin. 4. The process of claim 3 wherein said albumin is bovine serum albumin. 5. The process of claim 2 wherein the buffer is a phosphate or a GOOD buffer. 6. The process of claim 5 wherein the GOOD buffer is a HEPES or a TRIS buffer. 7. In a process for the determination of thyroxine-binding capacity in serum wherein a characteristic related to the capacity is determined by effective methods of determination, the improvement consisting essentially of calibrating the determination by means of two different serum-free standard solutions wherein a first standard solution contains a buffer system having a pH range of 6-8, a physiological amount of albumin, 10 to 60 .mu.g/ml thyroxine-binding globulin thereby corresponding to the highest thyroxine-binding capacity of human serum and a second standard solution contains a buffer system having a pH rang of 6-8, a physiological amount of albumin, 10 to 60 .mu.g/ml thyroxine-binding globulin and 100-500 ng/ml thyroxine thereby corresponding to the lowest thyroxine-binding capacity of human serum and adding each standard to a separate container having T4 antibody, further adding a predetermined amount of a reagent containing T4 and enzyme-conjugated T4 to each container, incubating the containers to permit competitive binding of the T4 and enzyme-conjugated T4 to thyroxine-binding protein and the anti-T4, removing unbound immunoreactants, adding substrate for the enzyme, determining, the amount of enzyme bound by measuring the colored product formed to produce a calibration curve from the values as determined above for the two standards, determining the amount of thyroxine-binding capacity by the effective method for determination with reference to the aforesaid calibration curve. 8. The process of claim 7 wherein said standard solutions contain bovine serum albumin.

Details for Patent 5,576,219

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Grifols Therapeutics Llc	ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5	albumin (human)	For Injection	101138	10/21/1942		2013-11-19
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		2013-11-19
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		2013-11-19
Grifols Biologicals Llc	ALBUTEIN	albumin (human)	Injection	102478	08/15/1978		2013-11-19
Instituto Grifols, S.a.	HUMAN ALBUMIN GRIFOLS	albumin (human)	Injection	103352	02/17/1995		2013-11-19