Production of Monoclonal Antibody against Prokaryotically Expressed G1 Protein of Bovine Ephemeral Fever Virus

محورهای موضوعی : Camel

ر. پسندیده ¹ (Department of Animal Science, Faculty of Animal and Food Science, Khuzestan Agricultural Science and Natural Resources University, Ahvaz, Iran)
م.ر. صیفی آباد شاپوری ² (Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran)
م.ت. بیگی نصیری ³ (Department of Animal Science, Faculty of Animal and Food Science, Khuzestan Agricultural Science and Natural Resources University, Ahvaz, Iran)

تاریخ دریافت : 1396/02/21 تاریخ پذیرش : 1396/05/09 تاریخ انتشار : 1397/12/10

کلید واژه: bovine ephemeral fever virus, hybridoma, monoclonal antibody, recombinant G1 protein,

چکیده مقاله :

Epitope-G1 of bovine ephemeral fever virus (BEFV) G glycoprotein has been genetically and antigenically conserved among various isolates of BEFV and only reacts with anti-BEFV neutralising antibodies. Therefore, it is a candidate antigen for development of the enzyme linked immunosorbent assay (ELISA) for serological identification bovine ephemeral fever (BEF)-infected animals. The aim of this study was to produce a monoclonal antibody (MAb) against recombinant G1 antigen expressed into Escherichia coli. For this purpose, somatic cell hybrids between SP2/0 myeloma cells and spleen cells derived from Balb/c mice immunized with maltose-binding protein (MBP)-G1 fusion protein were established. After three rounds of cloning, the stability of antibody secretion in the positive clones was confirmed by ELISA and the reactivity of the MAbs against recombinant G1 was verified by Western blot analysis. The specific MAbs produced against recombinant G1 antigen in this study could be used for establishing BEFV diagnostic experiments in the future.

چکیده انگلیسی:

اپیتوپ G1 از گلیکوپروتئین G ویروس تب بی ‏دوام گاوی (BEFV) از لحاظ ژنتیکی و آنتی ‏ژنیکی در بین جدایه‏ های مختلف از این ویروس حفاظت شده است و تنها با آنتی ‏بادی‏ های خنثی‏ کننده ضد این ویروس واکنش می‏ دهد. بنابراین این آنتی‏ژن، یک نامزد مناسب برای توسعه آزمون ایمونوسوربنت متصل به آنزیم (ELISA) برای شناسایی سرولوژیکی حیوانات آلوده است. هدف از این مطالعه، تولید آنتی ‏بادی مونوکلونال علیه آنتی‏ژن G1 نوترکیب بیان شده در اشرشیاکلی بود. به این منظور، هیبرید‏های سلولی سوماتیک بین سلول‏ های میلومای SP2/0 و سلول‏ های طحال حاصل از موش ‏های Balb/c ایمن شده با پروتئین هم جوش MBP-G1 ایجاد شد. پس از سه مرتبه کلونینگ، پایداری ترشح آنتی‏ بادی درکلون ‏های مثبت با روش الایزا و واکنش آنتی‏ بادی‏‏ های مونوکلونال علیه G1 نوترکیب توسط آزمایش وسترن بلات تأیید شد. آنتی‏ بادی‏ های اختصاصی تولید شده علیه آنتی ­ژن نوترکیب G1 ممکن است به ­منظور طراحی آزمایش‏ های تشخیصی برای ویروس BEF در آینده مناسب باشند.

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