Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces
A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional crosslinkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG a...
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| Veröffentlicht in: | Analytical biochemistry 1989-05, Vol.178 (2), p.408-413 |
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| container_title | Analytical biochemistry |
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| creator | Bhatia, Suresh K. Shriver-Lake, Lisa C. Prior, Kimberly J. Georger, Jacque H. Calvert, Jeffrey M. Bredehorst, Reinhard Ligler, Frances S. |
| description | A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional crosslinkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG antibody immobilized was determined to be in the range of 0.66 to 0.96 ng/mm
2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2–6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immonilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components. |
| doi_str_mv | 10.1016/0003-2697(89)90662-3 |
| format | Article |
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2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2–6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immonilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(89)90662-3</identifier><identifier>PMID: 2546467</identifier><identifier>CODEN: ANBCA2</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Antibodies ; Antigen-Antibody Reactions ; Biological and medical sciences ; Biotechnology ; Cross-Linking Reagents ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Immobilization of enzymes and other molecules ; Immobilization techniques ; Methods. Procedures. Technologies ; Molecular immunology ; Silicon Dioxide ; Sulfhydryl Compounds ; Techniques</subject><ispartof>Analytical biochemistry, 1989-05, Vol.178 (2), p.408-413</ispartof><rights>1989</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c483t-c64dc2e56b4ffe2b23fdaf228268c78549e1b38e262af49337be0a95e51316a3</citedby><cites>FETCH-LOGICAL-c483t-c64dc2e56b4ffe2b23fdaf228268c78549e1b38e262af49337be0a95e51316a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0003269789906623$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7295346$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2546467$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bhatia, Suresh K.</creatorcontrib><creatorcontrib>Shriver-Lake, Lisa C.</creatorcontrib><creatorcontrib>Prior, Kimberly J.</creatorcontrib><creatorcontrib>Georger, Jacque H.</creatorcontrib><creatorcontrib>Calvert, Jeffrey M.</creatorcontrib><creatorcontrib>Bredehorst, Reinhard</creatorcontrib><creatorcontrib>Ligler, Frances S.</creatorcontrib><title>Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional crosslinkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG antibody immobilized was determined to be in the range of 0.66 to 0.96 ng/mm
2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2–6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immonilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components.</description><subject>Antibodies</subject><subject>Antigen-Antibody Reactions</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cross-Linking Reagents</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Immobilization of enzymes and other molecules</subject><subject>Immobilization techniques</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular immunology</subject><subject>Silicon Dioxide</subject><subject>Sulfhydryl Compounds</subject><subject>Techniques</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9rFTEUxYMo9bX6DRRmIVIXo_k_yaYgRa1QcFPXIZO5oVdnJjWZEdpPb6bv8ZZ2Fbjndw-55xDyhtGPjDL9iVIqWq5td27sB0u15q14RnaMWt1SQe1zsjsiL8lpKb8oZUwqfUJOuJJa6m5H1p8FmhSb5RbT2C6QJ5z92BQc_Qyl8fPQ3EIdpx7jOocF0yaHnEoZcf4NuTQx5QanqRIjPviN2Az9vGCfBqwmdVD9MPimrDn6AOUVeRH9WOD14T0jN1-_3Fxetdc_vn2__HzdBmnE0gYth8BB6V7GCLznIg4-cm64NqEzSlpgvTDANfdRWiG6Hqi3ChQTTHtxRt7vbe9y-rNCWdyEJcC43ZbW4jpbszKGPwkyJUT1VBWUe_AxgQzR3WWcfL53jLqtFbdF7rbInbHusRUn6trbg__aTzAclw41VP3dQfcl-DFmPwcsR6zjVgmpK3axx6Bm9hchuxIQ5gADZgiLGxL-_x__ACOrquM</recordid><startdate>19890501</startdate><enddate>19890501</enddate><creator>Bhatia, Suresh K.</creator><creator>Shriver-Lake, Lisa C.</creator><creator>Prior, Kimberly J.</creator><creator>Georger, Jacque H.</creator><creator>Calvert, Jeffrey M.</creator><creator>Bredehorst, Reinhard</creator><creator>Ligler, Frances S.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><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>7T5</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19890501</creationdate><title>Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces</title><author>Bhatia, Suresh K. ; Shriver-Lake, Lisa C. ; Prior, Kimberly J. ; Georger, Jacque H. ; Calvert, Jeffrey M. ; Bredehorst, Reinhard ; Ligler, Frances S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c483t-c64dc2e56b4ffe2b23fdaf228268c78549e1b38e262af49337be0a95e51316a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Antibodies</topic><topic>Antigen-Antibody Reactions</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cross-Linking Reagents</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Immobilization of enzymes and other molecules</topic><topic>Immobilization techniques</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular immunology</topic><topic>Silicon Dioxide</topic><topic>Sulfhydryl Compounds</topic><topic>Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bhatia, Suresh K.</creatorcontrib><creatorcontrib>Shriver-Lake, Lisa C.</creatorcontrib><creatorcontrib>Prior, Kimberly J.</creatorcontrib><creatorcontrib>Georger, Jacque H.</creatorcontrib><creatorcontrib>Calvert, Jeffrey M.</creatorcontrib><creatorcontrib>Bredehorst, Reinhard</creatorcontrib><creatorcontrib>Ligler, Frances S.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bhatia, Suresh K.</au><au>Shriver-Lake, Lisa C.</au><au>Prior, Kimberly J.</au><au>Georger, Jacque H.</au><au>Calvert, Jeffrey M.</au><au>Bredehorst, Reinhard</au><au>Ligler, Frances S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1989-05-01</date><risdate>1989</risdate><volume>178</volume><issue>2</issue><spage>408</spage><epage>413</epage><pages>408-413</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><coden>ANBCA2</coden><abstract>A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional crosslinkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG antibody immobilized was determined to be in the range of 0.66 to 0.96 ng/mm
2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2–6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immonilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>2546467</pmid><doi>10.1016/0003-2697(89)90662-3</doi><tpages>6</tpages></addata></record> |
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| subjects | Antibodies Antigen-Antibody Reactions Biological and medical sciences Biotechnology Cross-Linking Reagents Fundamental and applied biological sciences. Psychology Fundamental immunology Immobilization of enzymes and other molecules Immobilization techniques Methods. Procedures. Technologies Molecular immunology Silicon Dioxide Sulfhydryl Compounds Techniques |
| title | Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces |
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