production and characterization of monoclonal antibody against recombinant virus coat protein cp42

Authors

naeimeh shibaei department of agricultural biotechnology, national institute of genetic engineering and biotechnology (nigeb), tehran, iran

jafar majidi department of immunology, faculty of medicine, tabriz university of medical sciences, tabriz, iran and immunology research center (irc), tabriz university of medical sciences, tabriz, iran

khadijeh razavi department of agricultural biotechnology, national institute of genetic engineering and biotechnology (nigeb), tehran, iran

ali asghar karkhane department of industrial and environmental biotechnology, national institute of genetic engineering and biotechnology (nigeb), tehran, iran

abstract

there are many studies related to the production of a elisa kit for diagnosing virus infections. however, production of most kits depends on purification of whole virus particles, which involves the use of costly equipment and reagents. the purpose of this study was to check out if the anti-cp42 antibodies could be used as a diagnostic assay for detection of grapevine fanleaf virus (gflv). in this study, recombinant gflv coat protein gene related to selected antigenic determinants was inserted into pet-28a bacterial expression vector and the construct (pet-28a cp42 ) was cloned into e. coli strain (de3). expressed protein was verified with western blotting assay by the use of commercially available anti-gflv antibody. the recombinant protein was purified using nickel–nitrilotriacetic acid (ni–nta) resin. balb/c mice were immunized with purified protein and splenocytes of hyperimmunized mice were fused with murine myeloma sp2/0 cells. positive hybridomas were selected by elisa using cp42 as coating antigen. the results showed that monoclonal antibody (mab) specific to cp42 has been successfully generated. efficiency of produced antibody was analyzed by elisa and western blotting assay using some confirmed grapevine samples. the infection was confirmed previously based on morphological features and elisa assay, performed using commercial anti-gflv antibody. the monoclonal antibody reacted with antigen in elisa and immunoblot method. our results demonstrated that anti recombinant cp42 monoclonal antibodies are able to diagnose whole virus in infected grapevine sample using elisa test.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Production of monoclonal antibody against recombinant NS3 protein of bovine viral diarrhea virus (NADL strain)

Bovine Viral Diarrhea virus (BVDV) is an important viral pathogen of cattle causing several clinical syndromes. There are usually no pathognomonic clinical signs of BVDV infection. Diagnostic investigations therefore rely on serological detection and virus isolation. Nonstructural protein 3 (NS3) as immunogenic protein of BVDV is genetically and antigenically conserved among different isolates....

full text

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

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 ...

full text

Production and Characterization of Monoclonal Antibody against Saffron Pollen Profilin, Cro s 2

Background: Allergy to Saffron (Crocus sativus) pollen has been described in people involved in processing of saffron flower stamens. Profilins have been identified as a pan-allergen in different plant pollens and foods. This molecule is an actin-binding pro-tein with a molecular weight of 12-16 kDa found in eukaryotic species. Objective: The aim of this study was to generate monoclonal antibod...

full text

preparation of polyclonal antibody against recombinant coat protein of cucumber mosaic virus isolate b13

cucumber mosaic virus (cmv) is one of widely-spread viruses of plants with the broadest host range encompassing over 1200 species. one major limiting factor for detection of the virus is unavailability of the virus-specific antibodies especially in developing countries. recombinant dna technology facilitates antibody preparation without requiring special equipment. in this study, coat protein (...

full text

Production of monoclonal antibody against recombinant NS3 protein of bovine viral diarrhea virus (NADL strain)

This study was conducted to investigate the prevalence of subclinical mastitis caused by Staphylococcus spp. in ewes in West-Azerbaijan province of Iran. Molecular characterization of isolated Staphylococcus spp. from diseased ewes were performed using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) and DNA sequencing of glyceraldehyde-3-phosphate deh...

full text

Prokaryotic Expression, Purification, and Polyclonal Antibody Production of a Truncated Recombinant Rabies Virus L Protein

Background: Rabies virus (RABV) is a deadly neurotropic virus that causes the disease of rabies in humans and animals. L protein is one of the large structural protein of rabies virus, which displays multiple enzymatic activities, and is required for viral transcription and replication. Objectives: A truncated L protein of Rabies virus is being cloned, expressed and purified to produce relevant...

full text

My Resources

Save resource for easier access later


Journal title:
iranian journal of allergy, asthma and immunology

جلد ۱۶، شماره ۱، صفحات ۶۰-۷۱

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023