purification of 146s antigen of foot-and-mouth disease (fmd) virus serotypes aby using the sucrose gradient procedure

نویسندگان

maryam shojaee department of biology, payame noor university of mashhad, mashhad, ir iran

saeid zibaei razi vaccine and serum research institute, agricultural research, education and extension organization (areeo) mashhad, ir iran; razi vaccine and serum research institute, agricultural research, education and extension organization (areeo) mashhad, ir iran. tel: +98-5138431780

چکیده

results the pellet of the 146s antigen of fmd virus serotype a was developed at a concentration of sucrose 50%. absorbance rate of the foot-and-mouth disease virus serotypes a at wavelengths of 240, 259 and 280 nm was 1.238, 1.573 and 1.157, respectively. moreover, the amount of 146s antigen at the same wavelengths was 163.416, 207.636 and 152.724 μg/ml, respectively. the amount of purified protein by nanodrop (nd-1000, the united states) was 0.275 mg/ml. the 146s antigen was observed with 26, 29 and 64 kda by sodium dodecyl sulfate polyacrylamide gel electrophoresis (sds-page) and was confirmed by the dot blot assay. objectives in this study, we aimed to introduce the sucrose gradient procedure as a convenient method for purification of 146s antigens. methods sucrose gradient procedure (20% - 50%) was used for purification of 146s antigen of foot-and-mouth disease virus serotypes a. then, preparation steps of the virus including concentration by polyethylene glycol, degreasing using trichloroethylene, centrifugation (30000 g for three hours) and washing the pellet using tris (0.05 m) were performed. spectrophotometer and nano-drop were used to measure the amount of the purified protein and purity evaluation, respectively. moreover, dot blot assay was used for the confirmation of 146s antigen. background foot-and-mouth disease (fmd) is an acute and contagious disease in domestic ruminants, which is currently the most economical viral disease that threatens the livestock industry. the virus that causes disease belongs to the aphthovirus genus from the family of picornaviridae. this family contains seven serotypes and is about 30 nanometers in diameter and has no external membrane, similar to other picornaviruses. conclusions the results exhibited that sucrose gradient procedure is a good method for purification of virus.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Comparison of two gel filtration chromatography resins for the purification of foot-and-mouth disease virus as a purified vaccine antigen

Introduction: Foot-and-mouth disease (FMD) is a highly contagious and economically devastating viral disease of livestock that is categorized in list A of animal diseases by the World Organisation for Animal Health (OIE). Vaccination is effective against FMD and the vaccine production centers largely use the industrial ultra-filtration and chromatography in order to remove the cellular proteins...

متن کامل

Receptor binding site-deleted foot-and-mouth disease (FMD) virus protects cattle from FMD.

Binding of foot-and-mouth disease virus (FMDV) to cells requires an arginine-glycine-aspartic acid (RGD) sequence in the capsid protein VP1. We have genetically engineered an FMDV in which these three amino acids have been deleted, producing a virus particle which is unable to bind to cells. Cattle vaccinated with these receptor binding site-deleted virions were protected from disease when chal...

متن کامل

Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development

The foot-and-mouth disease virus (FMDV) afflicts livestock in more than 80 countries, limiting food production and global trade. Production of foot-and-mouth disease (FMD) vaccines requires cytosolic expression of the FMDV 3C protease to cleave the P1 polyprotein into mature capsid proteins, but the FMDV 3C protease is toxic to host cells. To identify less-toxic isoforms of the FMDV 3C protease...

متن کامل

Assessment of the Immunogenicity of Foot and Mouth Disease Vaccine Produced by Razi Institute against Types of A13, A15 and O2010 of FMD Virus

Background and Aims: Foot-and-Mouth Disease (FMD) is a highly contagious infectious disease of livestock which has made a barrier to hygiene causing severe loss in livestock and their products. The aim of this study was the assessment of antibody response against foot and mouth disease virus  types A13, A15 , O2010, after injection of FMD vaccine candidate produced by Razi Institue. Mater...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
international journal of infection

جلد ۴، شماره ۱، صفحات ۰-۰

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023