Claims for Patent: 4,226,847

Summary for Patent: 4,226,847

Title:	Latex polymer sensitized with steroid-serum albumin conjugate, preparation and test therewith
Abstract:	Latex reagents for quantitative assay of a variety of steroid hormones or metabolites thereof which are contained in body fluid or excreted fluid of human beings comprise latex particles which are immunologically sensitized with a conjugate selected from a variety of steroid-serum albumin conjugates in which the steroid to be detected is bonded to serum albumin in a very small ratio ranging from 0.5 to 7 molecules per 1 molecule of the serum albumin used. Sensitization of latex particles with such a conjugate is performed using a limited amount dependent on the bonding ratio of steroid molecules per molecule of serum albumin in the conjugate and the particle size of the latex used. The latex reagents show very high sensitivity amounting to 0.2-0.04 nmole steroid equivalent/ml.
Inventor(s):	Ogasa; Katsuhiro (Yokohama, JP), Kuboyama; Morio (Tokyo, JP), Saito; Minoru (Komae, JP), Kudo; Tsutomu (Kawasaki, JP), Harada; Yoshitsugu (Yokohama, JP), Kawashiri; Akio (Fujimi, JP), Takahashi; Eiji (Tokyo, JP)
Assignee:	Morinaga Milk Industry Co., Ltd. (Tokyo, JP)
Application Number:	05/917,254
Patent Claims:	1. A method for preparation of a latex reagent containing latex particles sensitized with a steroid-serum albumin conjugate and suitable for immunochumical detection of steroid contained in human body fluid or excreted fluid, which comprises preparing steroid-serum albumin conjugate by reacting a steroid with a serum albumin in a ratio between 0.5 mole/mole serum albumin-7.0 mole/mole serum albumin, and preparing sensitized latex particles by sensitizing immunologically inert latex particles with said steroid-serum albumin conjugate. 2. The method of claim 1 wherein the sensitization of the latex particles with the conjugate is performed using a quantity of said conjugate which does not cause non-specific agglutination due to the sensitized latex particles themselves. 3. The method of claim 1, wherein said serum albumin is selected from the group consisting of bovine serum albumin, equine serum albumin, sheep serum albumin, rabbit serum albumin and human serum albumin. 4. The method of claim 1, wherein said latex particles are selected from the group consisting of polystyrene, polybutadiene and styrene-butadiene copolymer latex particles. 5. The method of claim 1 wherein said steroid used to prepare the conjugate can provoke an antigen-antibody reaction with the antibody for which a compound selected from the group consisting of estrogen and its metabolites, progesterone and its metabolites, androgen and its metabolites, and 17-hydroxycorticosterone and its metabolites can act as a hapten. 6. The method of claim 5, wherein said steroid used for preparation of the conjugate is selected from the group consisting of estrogen, progesterone, androgen, 17-hydroxycorticosterone and their metabolites. 7. The method of claim 1, wherein the particle size of said latex particles is within the range 0.234 .mu.m-0.721 .mu.m, and the quantity of said conjugate utilized for sensitization of said latex particles is within the range 0.3 .mu.mole/1 g latex particles-1.1 .mu.mole/1 g latex particles. 8. A latex reagent comprising latex particles sensitized with steroid-serum albumin conjugate and suitable for immunochemical detection of steroid contained in human body fluid or excreted fluid, wherein said conjugate has a steroid bonding number in the range 0.5-7.0 molecules per molecule of serum albumin, and said latex particles are sensitized with an effective amount of said conjugate to avoid non-specific agglutination but to cause agglutination due to antigen-antibody reaction between said latex reagent and the antibody for which the steroid to be detected may act as a hapten. 9. The latex reagent of claim 8, wherein the serum albumin moiety of said steroid-serum albumin conjugate is selected from the group consisting of bovine serum albumin, equine serum albumin, sheep serum albumin, rabbit serum albumin, and human serum albumin. 10. The latex reagent of claim 8, wherein said latex particles are selected from the group consisting of polystyrene, polybutadiene and styrene-butadiene copolymer latex particles. 11. The latex reagent of claim 8, wherein the steroid moiety of said steroid-serum albumin conjugate is a steroid capable of provoking an antigen-antibody reaction with the antibody for which a compound selected from the group consisting of estrogen and its metabolites, progesterone and its metabolites, androgen and its metabolites, and 17-hydroxycorticosterone and its metabolites can act as a hapten. 12. The latex reagent of claim 11, wherein said steroid is selected from the group consisting of estrogen, progesterone, androgen, 17-hydroxycorticosterone and their metabolites. 13. The latex reagent of claim 8, wherein the particle size of said latex is within the range 0.234 .mu.m-0.721 .mu.m, and the quantity of said conjugate utilized for sensitization of said latex particles is within the range 0.3 .mu.mole/1 g latex particles-1.1 .mu.mole/ 1 g latex particles. 14. In a method for the immunochemical detection of a steroid compound and/or metabolites thereof present in human body fluid or excreted fluid by forming a conjugate of a steroid and a protein material, forming sensitized latex particles by absorbing said conjugate on latex particles, obtaining an antiserum or antibody by injecting an animal with said conjugate and thereafter separating said antiserum or antibody from the blood of said animal and determining the presence of said steroid compound in a sample of body fluid by an agglutination reaction in the presence of said antiserum or antibody and said sensitized latex particles, the improvement comprising: increasing the sensitivity of said method by employing a conjugate having a steroid to serum albumin mole ratio ranging from 0.5 to 7.0. 15. The method of claim 14, wherein said serum albumin is selected from the group consisting of bovine serum albumin, equine serum albumin, sheep serum albumin, rabbit serum albumin and human serum albumin. 16. The method of claim 14, wherein said steroid used for preparation of the conjugate is selected from the group consisting of estrogen, progesterone, androgen, 17-hydroxycorticosterone and their metabolites.

Details for Patent 4,226,847

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		1997-10-07
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		1997-10-07
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		1997-10-07