{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:57:54Z","timestamp":1759971474323,"version":"build-2065373602"},"reference-count":23,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2021,7,3]],"date-time":"2021-07-03T00:00:00Z","timestamp":1625270400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2021,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>\u03b2\u2010Defensins are a family of conserved small cationic antimicrobial peptides with different significant biological functions.\u00a0The majority of mammalian \u03b2\u2010defensins are expressed in epididymis, and many of them are predicted to have post\u2010translational modifications. However, only a few of its members have been well studied due to the limitations of expressing and purifying bioactive proteins with correct post\u2010translational modifications efficiently.\u00a0Here we developed a novel Fc tagged lentiviral system and Fc tagged prokaryotic expression systems provided new options for \u03b2\u2010defensins expression and purification.\u00a0The novel lentiviral system contains a secretive signal peptide, an N\u2010terminal IgG Fc tag, a green fluorescent protein (GFP), and a puromycin selection marker to facilitate efficient expression and fast purification of \u03b2\u2010defensins by protein A magnetic or agarose beads.\u00a0It also enables stable and large\u2010scale expression of \u03b2\u2010defensins with regular biological activities and post\u2010translational modification.\u00a0Purified \u03b2\u2010defensins such as Bin1b and a novel human \u03b2\u2010defensin hBD129 showed antimicrobial activity, immuno\u2010regulatory activity, and expected post\u2010translational phosphorylation, which were not found in <jats:italic>Escherichia coli<\/jats:italic> (<jats:italic>E. coli<\/jats:italic>) in expressed form.\u00a0Furthermore, we successfully applied the novel system to identify mBin1b interacting proteins, explaining Bin1b in a better way. These results suggest that the novel lentiviral system is a powerful approach to produce correct post\u2010translational processed \u03b2\u2010defensins with bioactivities and is useful to identify their interacting proteins. This study has laid the foundation for future studies to characterize function and mechanism of novel \u03b2\u2010defensins.<\/jats:p>","DOI":"10.1002\/biot.202100023","type":"journal-article","created":{"date-parts":[[2021,5,30]],"date-time":"2021-05-30T13:00:50Z","timestamp":1622379650000},"source":"Crossref","is-referenced-by-count":1,"title":["An improved lentiviral system for efficient expression and purification of \u03b2\u2010defensins in mammalian cells"],"prefix":"10.1002","volume":"16","author":[{"given":"Shiraz","family":"Ahmed","sequence":"first","affiliation":[{"name":"Department of Endocrinology and Metabolism Shanghai Tenth People's Hospita, School of Medicine, Tongji University  No. 301 Middle Yanchang Road Shanghai 200072 China"},{"name":"State Key Laboratory of Molecular Biology Shanghai Key Laboratory of Molecular Andrology CAS Center for Excellence in Molecular Cell Science Shanghai Institute of Biochemistry and Cell Biology Chinese Academy of Sciences University of Chinese Academy of Sciences  No.320 Yueyang Road Shanghai 200031 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiang","family":"Tao","sequence":"additional","affiliation":[{"name":"Department of General dentistry, Shanghai Ninth People's Hospita, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases Shanghai Key Laboratory of Stomatology  No. 639 Zhizaoju Road Shanghai 200011 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miaochen","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of General dentistry, Shanghai Ninth People's Hospita, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases Shanghai Key Laboratory of Stomatology  No. 639 Zhizaoju Road Shanghai 200011 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yue","family":"Zhai","sequence":"additional","affiliation":[{"name":"Department of General dentistry, Shanghai Ninth People's Hospita, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases Shanghai Key Laboratory of Stomatology  No. 639 Zhizaoju Road Shanghai 200011 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenhai","family":"Liu","sequence":"additional","affiliation":[{"name":"Immunomic Therapeutics  No.15010 Broschart Road Rockville Maryland 20850 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Muthukumaran","family":"Jayachandran","sequence":"additional","affiliation":[{"name":"Department of Endocrinology and Metabolism Shanghai Tenth People's Hospita, School of Medicine, Tongji University  No. 301 Middle 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