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Affinity particle and affinity separation methodRelated Patent Categories: Liquid Purification Or Separation, Processes, ChromatographyAffinity particle and affinity separation method description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20070181503, Affinity particle and affinity separation method. Brief Patent Description - Full Patent Description - Patent Application Claims
TECHNICAL FIELD [0001] The present invention relates to affinity particles and an affinity separation method. More specifically, it relates to affinity particles utilizing inorganic particles and an affinity separation method that allows easy and highly precise separation of the target substance. The present invention is very useful in various separation, purification, and testing methods including latex agglutination methods and immunoprecipitation methods that allow easy and highly sensitive detection of the target substance. BACKGROUND ART [0002] Conventionally, column chromatography has been used for separation and purification of biological substances. However, column separation has some fatal problems as described in the following (1) to (3): [0003] (1) Many kinds of columns have to be used to obtain the target substance, resulting in a poor purification efficiency. [0004] (2) A verification test is required to make sure the target substance is contained in the fractionated ingredients, which means purification is time consuming. [0005] (3) Because of the large purification loss, a large quantity of the sample is required. [0006] On the other hand, for separation and purification of the target substances, affinity particles and affinity columns supporting ligands are used (Patent Document 1 and Patent Document 2). [0007] However, separation and purification using affinity columns have the following problems: [0008] (1) The desired target substance is not selectively separated. That is, in addition to the target substance captured by the ligand, unwanted substances are also adsorbed onto the column. [0009] (2) The capture efficiency is low, which means a large quantity of the liquid sample is required. [0010] The affinity separation method in which affinity particles are dispersed in a liquid sample for separation uses agarose and such (Non-patent Document 1), but this method has the following problems: [0011] (1) The desired target substance is not selectively separated. That is, in addition to the target substance captured by the ligand, unwanted target substances are also adsorbed onto the affinity particles. [0012] (2) The specific gravity is small, which makes the separation of the affinity particles difficult. [0013] (3) The carrier is easily disintegrated, which leads to poor durability. [0014] On the other hand, inorganic particles adsorb more substances than organic particles do, therefore those skilled in the art didn't think of using inorganic particles for affinity particles. [0015] Patent Document 1: Japanese Patent Publication H8-26076 [0016] Patent Document 2: Japanese Patent Laid-Open 2002-511141 bulletin [0017] Non-patent Document 1: Bioconjugate Chem.; 2002; 13(2); 163-166 DISCLOSURE OF INVENTION [Problem that the Present Invention Aims to Solve] [0018] The object of the present invention is to provide a groundbreaking affinity separation method that uses affinity particles utilizing inexpensive inorganic particles and is capable of separating the target substance easily and with high accuracy. The present invention is very useful in various separation, purification, and testing methods including latex agglutination methods and immunoprecipitation methods that allow easy and highly sensitive detection of the target substance. [Means to Solve the Problem] [0019] That is, the present invention provides affinity particles that are characterized by having phosphorylcholine groups represented by the following formula (1) covalently bonded onto the surface of inorganic particles. [0020] Also, the present invention provides affinity particles that are characterized by having phosphorylcholine groups represented by the following formula (1) covalently bonded onto the surface of inorganic particles and also by having reactive groups or adsorptive groups, which are capable of bonding with ligands having specific affinity with a certain target substance, covalently bonded or adsorbed onto the surface of inorganic particles. [0021] Furthermore, the present invention provides affinity particles that are characterized by having phosphorylcholine groups represented by the following formula (1) covalently bonded onto the surface of inorganic particles and also by having ligands having specific affinity with a certain target substance covalently bonded or adsorbed onto the surface of inorganic particles. [Chemical formula 6] [0022] Also, the present invention provides the aforementioned affinity particles wherein said inorganic particles are selected from a group consisting of silica, titanium oxide, zinc flower, alumina, iron oxide, talc, mica, sericite, and gold colloid, and having an average particle size of 20 nm to 500 .mu.m and a specific gravity of 1.0 g/cm.sup.3 or higher. [0023] Furthermore, the present invention provides the aforementioned affinity particles wherein said ligands are one, two, or more types of ligands chosen from a group consisting of various antibodies, antigens, enzymes, substrates, lectin, receptors, peptides, DNA, RNA, aptamers, protein A, protein G, avidin, biotin, chelating compounds, and various metal ions. [0024] Also, the present invention provides a method of affinity separation of a target substance by using inorganic particles that includes (1) a first process whereby arbitrary ligands are bonded to the affinity particles of claim 1 or 2, (2) a second process whereby the affinity particles prepared in the first process are dispersed in a liquid sample containing a target substance selectively captured by the arbitrary ligands, and (3) a third process whereby the target substance captured is recovered from the affinity particles. [0025] Furthermore, the present invention provides a method of affinity separation of a target substance by using inorganic particles that includes (1) a first process whereby the affinity particles of claim 3 are dispersed in a liquid sample containing a target substance selectively captured by the arbitrary ligands, and (2) a second process whereby the target substance captured is recovered from the affinity particles. When the affinity particles of the present invention are used for detection of antibodies and protein, such as in the immunoprecipitation method and the latex agglutination method, the recovery process (2) is not required; detection can be done easily by visually observing changes in the dispersion state. [Effects of the Invention] [0026] The affinity particles of the present invention use ligands to capture only a certain target substance (the substance desired to be separated) and suppresses adsorption of other substances onto the particles, resulting in a very high separation selectivity. They also exhibit superior dispersion properties and make separation from liquid samples very easy, which makes it possible to separate the target substance easily and with high accuracy by using inexpensive inorganic powder particles for affinity particles. Continue reading about Affinity particle and affinity separation method... Full patent description for Affinity particle and affinity separation method Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Affinity particle and affinity separation method patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. 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