Synthetic resins or natural rubbers -- part of the class 520 ser – Synthetic resins – Mixing of two or more solid polymers; mixing of solid...
Patent
1982-07-20
1994-03-08
Jagannathan, Vasu S.
Synthetic resins or natural rubbers -- part of the class 520 ser
Synthetic resins
Mixing of two or more solid polymers; mixing of solid...
525302, 525304, 525305, 525308, 436 67, C08F26504, G01N 3372
Patent
active
052928187
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a production method of a carrier appropriate for cation exchange liquid chromatography and a method for determining glycosylated hemoglobins utilizing the carrier obtained by the production method.
BACKGROUND ART
A liquid chromatography is used for a separation or a detection of various materials, and especially for a separation or a detection of a hydrophilic material, for example, for a separation of protein from a biological sample, an ion exchange chromatography is used. Ion exchange chromatography is a method in which a carrier having an ion exchange group is utilized for separating materials depending upon a difference in the strength of an ionic bond of the material to be separated to the carrier. A weak cation exchange chromatography utilizing a carrier having a carboxyl group as the ion exchange group is effective for an analysis of protein or peptide.
For example, glycosylated hemoglobins in blood are measured with ion exchange chromatography utilizing such a carrier.
The glycosylated hemoglobins are formed by a non-enzymatical reaction of hemoglobin in an erythrocyte with glucose in the blood. Since a measurement of glycosylated hemoglobins reveals an average concentration of glucose in the blood, the measurement of the glycosylated hemoglobins is widely used for the diagnosis of diabetes. The glycosylated hemoglobins are now determined by a high performance liquid chromatography (hereinafter referred to as the "HPLC") mainly utilizing the above-mentioned carrier. The HPLC provides rapid measurement as compared with conventional column chromatography, electrophoresis, colorimetric analysis and the like.
An organic polymer carrier or an inorganic carrier is generally used as a carrier for weak cation exchange chromatography. A carrier gel of organic polymers used most frequently is a carrier in which carboxyl groups are introduced on a surface of a particle of a styrene-divinylbenzene crosslinked copolymer. Such a carrier is obtained by introducing carboxyl groups to a particle of a styrene-divinylbenzene crosslinked copolymer by a chemical reaction. For example, alkyl halide is introduced to a benzene ring of styrene in the copolymer by treating the particle of the copolymer with chloromethyl ether and the like, and the resultant material is hydrolyzed and oxidized, thereby introducing the carboxyl groups.
An example of the organic polymer carrier, besides the above-mentioned gel, includes particles of a crosslinked copolymer of styrene, divinylbenzene and a monomer having a carboxyl group. Moreover, Japanese Laid-Open Patent Publication No. 58-221164 discloses a carrier made of a copolymer of an acrylate or a methacrylate such as tetramethylolmethane triacrylate and acrylic acid or methacrylic acid. Such a carrier is generally prepared in a method disclosed in the foregoing Japanese Laid-Open Patent Publication No. 58-221164: for example, the crosslinked copolymer particles are prepared by adding a polymerization initiator to a mixture of a crosslinkable monomer and a monomer having a carboxyl group for a suspension polymerization. Alternatively, particles obtained by copolymerizing styrene, divinylbenzene and a monomer having a functional group which can be converted into a carboxyl group by a hydrolytic reaction (hereinafter referred to as the "hydrolyzable group") and then converting the functional group into a carboxyl group by a hydrolytic reaction can be used. It is necessary to increase the degree of the crosslink in order to improve the pressure resistance of such a carrier. However, when the degree of the crosslink is increased, the hydrophobicity of the gel is also increased since a crosslinked portion is hydrophobic, resulting in a nonspecific adsorption of protein. Therefore, the amount of a crosslinking agent to be used is limited, and it is difficult to obtain a satisfactory pressure resistance. Moreover, since the carrier obtained in the above-mentioned manner contains carboxyl groups in the entire copolymer particle, the carri
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Kawabe Toshiki
Oishi Kazuyuki
Takahara Makoto
Takechi Masahiro
Yamazaki Kazutoshi
Jagannathan Vasu S.
Sekisui Kagaku Kogyo Kabushiki Kaisha
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