{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T10:56:03Z","timestamp":1790592963641,"version":"4.1.0"},"reference-count":31,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22004097"],"award-info":[{"award-number":["22004097"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Separation Science"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Immobilization of functional proteins onto solid surfaces is pivotal to propel the advancement of biomaterials in both biomedical and industrial technologies. However, the process has been hindered when it comes to G protein\u2010coupled receptors due to the homogeneity of the seven transmembrane proteins. Herein, we exploited the efficient ultrahigh affinity of colicin E7 (CL7) and its cognate immunity protein IM7 for conjugation of functional protein onto macro\u2010porous silica gel and established a robust chromatographic system for separation of compounds from complex matrices. Benefiting from the ultrahigh affinity nature of the protein pair, conjugation of \u03b1\n                    <jats:sub>1A<\/jats:sub>\n                    \u2010adrenoceptor (\u03b1\n                    <jats:sub>1A<\/jats:sub>\n                    \u2010AR) was achieved at ambient conditions in less than 20\u00a0min with high specificity and stability. Feasibility of the platform was tested by determining the binding parameters between drugs and receptor under various conditions (different pHs and concentration of methanol). Under the desired condition, carvedilol gave the association constant of (11.56\u00b10.7) \u00d7\u00a010\n                    <jats:sup>5<\/jats:sup>\n                    M\n                    <jats:sup>\u22121<\/jats:sup>\n                    indicating immobilization of \u03b1\n                    <jats:sub>1A<\/jats:sub>\n                    \u2010AR via this method still enhances its binding activity to drugs. Furthermore, stachydrine and leonurine were obtained as the bioactive compounds targeting the immobilized \u03b1\n                    <jats:sub>1A<\/jats:sub>\n                    \u2010AR. Our study is expected to offer a versatile platform to achieve effective and specific immobilization of other functional proteins, thus enabling easy separation of the target compound from complex systems.\n                  <\/jats:p>","DOI":"10.1002\/jssc.70524","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T10:17:18Z","timestamp":1788776238000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Construction of\n                    <i>\u03b1<\/i>\n                    <sub>1A<\/sub>\n                    \u2010Adrenoceptor\u2010Based Stationary Phase Triggered by the Ultrahigh Affinity Protein Pair Colicin E7 Dnase \/IM7 for Targeted Bioactive Compound Screening in\n                    <i>Herba leonuri<\/i>"],"prefix":"10.1002","volume":"49","author":[{"given":"Xiaofang","family":"Qiao","sequence":"first","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China Northwest University  Xi'an China"},{"name":"Henan Provincial Drug Review and Inspection Center (Henan Provincial Vaccine Inspection Center)  Zhengzhou Henan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haosen","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China Northwest University  Xi'an China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China Northwest University  Xi'an China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Liu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China Northwest University  Xi'an China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Guo","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China Northwest University  Xi'an China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China Northwest University  Xi'an China"},{"name":"Yunnan Characteristic Plant Extraction Laboratory Co. Ltd  Kunming Yunnan China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,7]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010019\u201010696\u2010x"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsabm.2c00709"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.9b02096"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.4c00509"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.3c06930"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.R120.001936"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010021\u201003258\u2010z"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200801711"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc060125e"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/cr8004668"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cclet.2024.109792"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsami.1c14670"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc200396r"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc500443h"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.3390\/s24092805"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc500347b"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.3751"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4080"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2015.07.014"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.13.6437"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/acssuschemeng.1c06647"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2008.11.026"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2025.124478"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2019.105524"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2009.267"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2010.10.003"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja01238a017"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/prp2.602"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216\u2010011\u20105026\u2010z"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1002\/jms.4978"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2013.12.052"}],"container-title":["Journal of Separation Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jssc.70524","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jssc.70524","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jssc.70524","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T10:01:36Z","timestamp":1790589696000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jssc.70524"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9]]},"references-count":31,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2026,9]]}},"alternative-id":["10.1002\/jssc.70524"],"URL":"https:\/\/doi.org\/10.1002\/jssc.70524","archive":["Portico"],"relation":{},"ISSN":["1615-9306","1615-9314"],"issn-type":[{"value":"1615-9306","type":"print"},{"value":"1615-9314","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9]]},"assertion":[{"value":"2026-06-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-28","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-07","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70524"}}