Rapid Purification of Immunoglobulin G Using a Protein A-immobilized Monolithic Spin Column with Hydrophilic Polymers
A rapid purification method was developed for antibody production in Chinese hamster ovary (CHO) cells using a Protein A-immobilized monolithic silica spin column with hydrophilic polymers. Monolithic silica modified with copolymers of 2-hydroxyethylmethacrylate (HEMA) and glycidyl methacrylate (GMA...
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| Veröffentlicht in: | Analytical Sciences 2021/07/10, Vol.37(7), pp.985-990 |
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| creator | OTA, Shigenori YUI, Yuko SATO, Tsutomu YOSHIMOTO, Noriko YAMAMOTO, Shuichi |
| description | A rapid purification method was developed for antibody production in Chinese hamster ovary (CHO) cells using a Protein A-immobilized monolithic silica spin column with hydrophilic polymers. Monolithic silica modified with copolymers of 2-hydroxyethylmethacrylate (HEMA) and glycidyl methacrylate (GMA) showed lower non-specific protein absorption than that modified with a silane reagent. The epoxy group of GMA was converted to an amino group, and Protein A was modified by the coupling reagent. The amount of immobilized Protein A was controlled by changing the ratio of GMA to HEMA and the mesopore size of monolith. A modified monolith disk was fixed to a spin column for rapid antibody purification. The linear curves (for the antibody concentrations over 10 – 300 μg/mL) had a correlation coefficient of >0.999. Our column had various analytical advantages over previously reported columns, including a shorter preparation time ( |
| doi_str_mv | 10.2116/analsci.20P378 |
| format | Article |
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Monolithic silica modified with copolymers of 2-hydroxyethylmethacrylate (HEMA) and glycidyl methacrylate (GMA) showed lower non-specific protein absorption than that modified with a silane reagent. The epoxy group of GMA was converted to an amino group, and Protein A was modified by the coupling reagent. The amount of immobilized Protein A was controlled by changing the ratio of GMA to HEMA and the mesopore size of monolith. A modified monolith disk was fixed to a spin column for rapid antibody purification. The linear curves (for the antibody concentrations over 10 – 300 μg/mL) had a correlation coefficient of >0.999. Our column had various analytical advantages over previously reported columns, including a shorter preparation time (<10 min) and smaller sample volumes for purification with Protein A-immobilized agarose.</description><identifier>ISSN: 0910-6340</identifier><identifier>EISSN: 1348-2246</identifier><identifier>DOI: 10.2116/analsci.20P378</identifier><identifier>PMID: 33281136</identifier><language>eng</language><publisher>Singapore: The Japan Society for Analytical Chemistry</publisher><subject>affinity chromatography ; Analytical Chemistry ; Animals ; Antibodies ; Chemistry ; CHO Cells ; Copolymers ; Correlation coefficient ; Correlation coefficients ; Cricetinae ; Cricetulus ; Hydrophilicity ; Hydrophobic and Hydrophilic Interactions ; IgG antibody ; Immunoglobulin G ; Monolithic silica ; non-specific adsorption ; Original Paper ; Polyhydroxyethyl methacrylate ; Polymers ; Protein A ; Protein purification ; Proteins ; Purification ; Reagents ; Silica ; Silicon dioxide</subject><ispartof>Analytical Sciences, 2021/07/10, Vol.37(7), pp.985-990</ispartof><rights>2021 by The Japan Society for Analytical Chemistry</rights><rights>The Japan Society for Analytical Chemistry 2021</rights><rights>Copyright Japan Science and Technology Agency 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c484t-e4887a355448285367063bb0dc86cfecfd641f1bd6cecb85d61a52cdc3e0cc303</citedby><cites>FETCH-LOGICAL-c484t-e4887a355448285367063bb0dc86cfecfd641f1bd6cecb85d61a52cdc3e0cc303</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.2116/analsci.20P378$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.2116/analsci.20P378$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,1883,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33281136$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>OTA, Shigenori</creatorcontrib><creatorcontrib>YUI, Yuko</creatorcontrib><creatorcontrib>SATO, Tsutomu</creatorcontrib><creatorcontrib>YOSHIMOTO, Noriko</creatorcontrib><creatorcontrib>YAMAMOTO, Shuichi</creatorcontrib><title>Rapid Purification of Immunoglobulin G Using a Protein A-immobilized Monolithic Spin Column with Hydrophilic Polymers</title><title>Analytical Sciences</title><addtitle>ANAL. SCI</addtitle><addtitle>Anal Sci</addtitle><description>A rapid purification method was developed for antibody production in Chinese hamster ovary (CHO) cells using a Protein A-immobilized monolithic silica spin column with hydrophilic polymers. Monolithic silica modified with copolymers of 2-hydroxyethylmethacrylate (HEMA) and glycidyl methacrylate (GMA) showed lower non-specific protein absorption than that modified with a silane reagent. The epoxy group of GMA was converted to an amino group, and Protein A was modified by the coupling reagent. The amount of immobilized Protein A was controlled by changing the ratio of GMA to HEMA and the mesopore size of monolith. A modified monolith disk was fixed to a spin column for rapid antibody purification. The linear curves (for the antibody concentrations over 10 – 300 μg/mL) had a correlation coefficient of >0.999. Our column had various analytical advantages over previously reported columns, including a shorter preparation time (<10 min) and smaller sample volumes for purification with Protein A-immobilized agarose.</description><subject>affinity chromatography</subject><subject>Analytical Chemistry</subject><subject>Animals</subject><subject>Antibodies</subject><subject>Chemistry</subject><subject>CHO Cells</subject><subject>Copolymers</subject><subject>Correlation coefficient</subject><subject>Correlation coefficients</subject><subject>Cricetinae</subject><subject>Cricetulus</subject><subject>Hydrophilicity</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>IgG antibody</subject><subject>Immunoglobulin G</subject><subject>Monolithic silica</subject><subject>non-specific adsorption</subject><subject>Original Paper</subject><subject>Polyhydroxyethyl methacrylate</subject><subject>Polymers</subject><subject>Protein A</subject><subject>Protein purification</subject><subject>Proteins</subject><subject>Purification</subject><subject>Reagents</subject><subject>Silica</subject><subject>Silicon dioxide</subject><issn>0910-6340</issn><issn>1348-2246</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kU9vEzEQxS0EoqFw5YgsceGyqf-t1xyrAG2lIiKgZ8trexNHXnuxdwXpqXe-JZ8ERxuChMTFI8383htrHgAvMVoSjPmFCspn7ZYErWkjHoEFpkxUhDD-GCzQW4wqThk6A89y3iGEiSDkKTijlAiMKV-AH5_V4AxcT8l1TqvRxQBjB2_6fgpx42M7eRfgFbzLLmyggusUR1s6l9Wvh5-u72PrvLu3Bn6MIXo3bp2GX4YCrKKf-gC_lxa83psUh20hNVxHv-9tys_Bk6583b441nNw9-H919V1dfvp6mZ1eVtpJthYWSZEo2hdMyaIqClvEKdti4wWXHdWd4Yz3OHWcG11K2rDsaqJNppapDVF9By8mX2HFL9NNo-yd1lb71WwccqyXKoRjNSUFfT1P-guTulwYEnquiGcllKo5UzpFHNOtpNDcr1Ke4mRPGQij5nIOZMieHW0ndremhP-J4QCXMxALqOwsenv3v9avpsVuzyqjT1ZqjQ67e0Jp41sDs8sO431ViVpA_0Nnm6z-Q</recordid><startdate>20210710</startdate><enddate>20210710</enddate><creator>OTA, Shigenori</creator><creator>YUI, Yuko</creator><creator>SATO, Tsutomu</creator><creator>YOSHIMOTO, Noriko</creator><creator>YAMAMOTO, Shuichi</creator><general>The Japan Society for Analytical Chemistry</general><general>Springer Nature Singapore</general><general>Japan Science and Technology Agency</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SE</scope><scope>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>FR3</scope><scope>H8G</scope><scope>JG9</scope><scope>L7M</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20210710</creationdate><title>Rapid Purification of Immunoglobulin G Using a Protein A-immobilized Monolithic Spin Column with Hydrophilic Polymers</title><author>OTA, Shigenori ; 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| subjects | affinity chromatography Analytical Chemistry Animals Antibodies Chemistry CHO Cells Copolymers Correlation coefficient Correlation coefficients Cricetinae Cricetulus Hydrophilicity Hydrophobic and Hydrophilic Interactions IgG antibody Immunoglobulin G Monolithic silica non-specific adsorption Original Paper Polyhydroxyethyl methacrylate Polymers Protein A Protein purification Proteins Purification Reagents Silica Silicon dioxide |
| title | Rapid Purification of Immunoglobulin G Using a Protein A-immobilized Monolithic Spin Column with Hydrophilic Polymers |
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