{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T18:07:25Z","timestamp":1783102045823,"version":"3.54.6"},"reference-count":27,"publisher":"American Society for Microbiology","issue":"10","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clin. Vaccine Immunol."],"published-print":{"date-parts":[[2011,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Rabies is a fatal zoonotic disease of serious public health and economic significance worldwide. The rabies virus glycoprotein (RVG) has been the major target for subunit vaccine development, since it harbors domains responsible for induction of virus-neutralizing antibodies, infectivity, and neurovirulence. The glycoprotein (G) was cloned using the baculovirus expression vector system (BEVS) and expressed in<jats:named-content content-type=\"genus-species\">Spodoptera frugiperda<\/jats:named-content>(Sf-9) cells. In order to obtain a soluble form of G suitable for experimentation in mice, 18 different combinations of buffers and detergents were evaluated for their ability to solubilize the insect cell membrane-associated G. The combination that involved 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) detergent in lysis buffer 1, formulated with Tris, NaCl, 10% dimethyl sulfoxide (DMSO), and EDTA, gave the highest yield of soluble G, as evidenced by the experimental data. Subsequently, several other parameters, such as the concentration of CHAPS and the duration and temperature of the treatment for the effective solubilization of G, were optimized. The CHAPS detergent, buffered at a concentration of 0.4% to 0.7% (wt\/vol) at room temperature (23 to 25\u00b0C) for 30 min to 1 h using buffer 1, containing 10% DMSO, resulted in consistently high yields. The G solubilized using CHAPS detergent was found to be immunogenic when tested in mice, as evidenced by high virus-neutralizing antibody titers in sera and 100% protection upon virulent intracerebral challenge with the challenge virus standard (CVS) strain of rabies virus. The results of the mice study indicated that G solubilized with CHAPS detergent retained the immunologically relevant domains in the native conformation, thereby paving the way for producing a cell-free and efficacious subunit vaccine.<\/jats:p>","DOI":"10.1128\/cvi.05258-11","type":"journal-article","created":{"date-parts":[[2011,8,4]],"date-time":"2011-08-04T00:58:00Z","timestamp":1312419480000},"page":"1673-1679","source":"Crossref","is-referenced-by-count":23,"title":["Expression and Solubilization of Insect Cell-Based Rabies Virus Glycoprotein and Assessment of Its Immunogenicity and Protective Efficacy in Mice"],"prefix":"10.1128","volume":"18","author":[{"given":"R.","family":"Ramya","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B.","family":"Mohana Subramanian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V.","family":"Sivakumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. 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J."},{"key":"key20180919073535_B2","doi-asserted-by":"crossref","first-page":"1917","DOI":"10.1089\/104303401753153965","article-title":"Preexposure efficacy of a novel combination DNA and inactivated rabies virus vaccine","volume":"12","author":"Biswas","year":"2001","journal-title":"Hum. Gene Ther."},{"key":"key20180919073535_B3","first-page":"45","article-title":"Structure and function of rabies virus glycoprotein","volume":"40","author":"Dietzschold","year":"1978","journal-title":"Dev. Biol. Stand."},{"key":"key20180919073535_B4","doi-asserted-by":"crossref","first-page":"e515","DOI":"10.1371\/journal.pntd.0000515","article-title":"Novel vaccines to human rabies","volume":"3","author":"Ertl","year":"2009","journal-title":"PLoS Negl. Trop. Dis."},{"key":"key20180919073535_B5","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1016\/0264-410X(93)90379-C","article-title":"Oral vaccination of raccoons (Procyon lotor) with baculovirus-expressed rabies virus glycoprotein","volume":"11","author":"Fu","year":"1993","journal-title":"Vaccine"},{"key":"key20180919073535_B6","doi-asserted-by":"crossref","first-page":"32403","DOI":"10.1074\/jbc.R100031200","article-title":"Detergents as tools in membrane biochemistry","volume":"276","author":"Garavito","year":"2001","journal-title":"J. Biol. Chem."},{"key":"key20180919073535_B7","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.vetmic.2005.04.007","article-title":"Immunogenic and antigenic properties of recombinant soluble glycoprotein of rabies virus","volume":"108","author":"Gupta","year":"2005","journal-title":"Vet. Microbiol."},{"key":"key20180919073535_B8","doi-asserted-by":"crossref","first-page":"10453","DOI":"10.1021\/jp073113e","article-title":"Modulating the structure and properties of cell membranes: the molecular mechanism of action of dimethyl sulfoxide","volume":"111","author":"Gurtovenko","year":"2007","journal-title":"J. Phys. Chem. B"},{"key":"key20180919073535_B9","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1016\/0076-6879(79)56066-2","article-title":"Properties of detergents","volume":"56","author":"Helenius","year":"1979","journal-title":"Methods Enzymol."},{"key":"key20180919073535_B10","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/0076-6879(90)82023-U","article-title":"Removal of detergents from membrane proteins","volume":"182","author":"Hjelmeland","year":"1990","journal-title":"Methods Enzymol."},{"issue":"4","key":"key20180919073535_B11","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1111\/j.1745-7254.2005.00078.x","article-title":"Baculovirus as a highly efficient expression vector in insect and mammalian cells","volume":"26","author":"Hu","year":"2005","journal-title":"Acta Pharmacol. Sin."},{"key":"key20180919073535_B12","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1007\/s00705-010-0909-4","article-title":"Construction and immunogenicity of a recombinant pseudotype baculovirus expressing the glycoprotein of rabies virus in mice","volume":"156","author":"Huang","year":"2011","journal-title":"Arch. Virol."},{"key":"key20180919073535_B13","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1007\/BF01309439","article-title":"Characterization of rabies glycoprotein expressed in yeast","volume":"128","author":"Klepfer","year":"1993","journal-title":"Arch. Virol."},{"key":"key20180919073535_B14","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1111\/j.1432-1033.1983.tb07668.x","article-title":"Topography of the total protein population from cultured cells upon fractionation by chemical extractions","volume":"135","author":"Lenstra","year":"1983","journal-title":"Eur. J. Biochem."},{"key":"key20180919073535_B15","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1016\/S0076-6879(09)63035-4","article-title":"Purification of membrane proteins","volume":"463","author":"Lin","year":"2009","journal-title":"Methods Enzymol."},{"key":"key20180919073535_B16","first-page":"946","article-title":"Preparation of mammalian plasma membranes by aqueous two-phase partitioning","volume":"7","author":"Morre","year":"1989","journal-title":"Biotechniques"},{"key":"key20180919073535_B17","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.biologicals.2005.07.004","article-title":"A simple immune-capture ELISA to estimate rabies viral glycoprotein antigen in vaccine manufacture","volume":"34","author":"Nagarajan","year":"2006","journal-title":"Biologicals"},{"key":"key20180919073535_B18","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/0042-6822(89)90551-5","article-title":"Immunogenic and protective properties of rabies virus glycoprotein expressed by baculovirus vectors","volume":"173","author":"Prehaud","year":"1989","journal-title":"Virology"},{"key":"key20180919073535_B19","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1007\/s11248-009-9278-8","article-title":"Expression of rabies virus G protein in carrots (Daucus carota)","volume":"18","author":"Rojas-Anaya","year":"2009","journal-title":"Transgenic Res."},{"key":"key20180919073535_B20","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/S0264-410X(98)00196-0","article-title":"Studies on the structures and antigenic properties of rabies virus glycoprotein analogues produced in yeast cells","volume":"17","author":"Sakamoto","year":"1999","journal-title":"Vaccine"},{"key":"key20180919073535_B21","first-page":"181","article-title":"Rapid fluorescent focus inhibition test for determining rabies virus neutralizing antibodies","volume-title":"Laboratory techniques in rabies","author":"Smith","year":"1996","edition":"4th ed."},{"key":"key20180919073535_B22","first-page":"263","article-title":"The rabies virus genome: an overview","volume":"60","author":"Tordo","year":"1993","journal-title":"Onderstepoort J. 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