Production and Easy One-Step Purification of Bluetongue Recombinant VP7 from Infected Sf9 Supernatant for an Immunoenzymatic Assay (ELISA)

被引：3

作者：

Ulisse, S. ^{[1
]}

Iorio, M. ^{[1
]}

Armillotta, G. ^{[1
]}

Laguardia, C. ^{[1
]}

Testa, L. ^{[1
]}

Capista, S. ^{[1
]}

Centorame, P. ^{[1
]}

Traini, S. ^{[1
]}

Serroni, A. ^{[1
]}

Monaco, F. ^{[1
]}

Caporale, M. ^{[1
]}

Mercante, M. T. ^{[1
]}

Di Ventura, M. ^{[1
]}

机构：

[1] Ist Zooprofilatt Sperimentale Abruzzo & Molise G, Teramo, Italy

来源：

MOLECULAR BIOTECHNOLOGY | 2021年 / 63卷 / 01期

关键词：

BTV; Recombinant VP7; Baculovirus; Supernatant; Affinity chromatography; ELISA; BACULOVIRUS EXPRESSION SYSTEM; LINKED-IMMUNOSORBENT-ASSAY; CAPSID PROTEIN; RAPID METHOD; VIRUS BTV; ANTIGEN; ARGININE; TITER; IDENTIFICATION; PARTICLES;

D O I：

10.1007/s12033-020-00282-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Bluetongue (BT) is non-contagious, vector-borne viral disease of domestic and wild ruminants, transmitted by midges (Culicoidesspp.) and is caused by Bluetongue virus (BTV). BTV is the type species of theOrbivirusgenus within theReoviridaefamily and possesses a genome consisting of 10 double-stranded RNA segments encoding 7 structural and 4 nonstructural proteins. Viral Protein 7 (VP7) is the major sera group-specific protein and is a good antigen candidate for immunoenzymatic assays for the BT diagnosis. In our work, BTV-2 recombinant VP7 (BTV-2 recVP7), expressed inSpodoptera frugiperda(Sf9) cells using a baculovirus system, was produced and purified by affinity chromatography from the supernatant of infected cell culture. The use of the supernatant allowed us to obtain a high quantity of recombinant protein with high purity level by an easy one-step procedure, rather than the multistep purification from the pellet. RecVP7-BTV2 was detected using a MAb anti-BTV in Western blot and it was used to develop an immunoenzymatic assay.

引用

页码：40 / 52

页数：13

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