For immunization, a two-month male rabbit (2 Kg weight) was selected 120 μL of purified antigen 146S along with 880 μL Tris buffer 0.05 M and 1 mL of the CFA adjuvant in a total volume of 2 mL was prepared as an injection suspension. Laboratory animals were provided by production and research center of Razi institute. Immunizing a White New Zealand Rabbit to Produce Specific Antibody The virus was inactivated by ethylene amine double method and purified by sucrose gradient procedure.ģ.2. The current study aimed to introduce ion exchange chromatography as a convenient method to purify specific antibody against 146S antigen.įoot-and-mouth disease virus serotype A was provided by research and production of foot-and-mouth disease vaccine center of Razi institute, Tehran, Iran. The current study aimed to introduce ion exchange chromatography as a convenient method to purify specific antibody against 146S antigen of FMDV serotype A. At present, reverse transcription polymerase chain reaction (RT-PCR) along with ELISA and cell culture are used for early detection and determination of the type of FMD virus in the world reference laboratory (WRL) ( 24). Complement fixation test (CFT) was widely used to detect the FMD virus, but it was alternated by ELISA due to its sensitivity and assessment of a lot of samples. Since FMD is on top of the world organization for animal health (OIE) list, early and rapid virus detection has a great importance. One case reported that FMD killed two children supposedly due to infected milk in England in 1884 ( 23). FMD in humans has symptoms including malaise, fever, vomiting, red ulcerative lesions of the oral tissue and sometimes vesicular lesions of the skin. Moreover, only a few other cases were recorded in countries of the continental Europe, Africa, and South America. The last confirmed human case happened in 1966 ( 20- 22) in the United Kingdom. The virus that causes FMD cannot spread to humans via consumption of infected meat because it is sensitive to stomach acid. Humans are very rarely infected with FMD through contact with polluted animals. A main antigenic site on FMDV is identified around amino acid residues 140 to 160 of VP1, the extreme carboxy terminus (residues 200 to 213) of that protein likely contribute to the antigenicity ( 16- 19). Of the four structural proteins, only VP1 elicits neutralizing antibodies in animals ( 12- 15). The virus exhibits a great antigenic variability extensively characterized in tissue culture and in the field ( 8- 11). Similar to other picornaviruses, FMDV has a single stranded positive RNA molecule ( 6) encapsidated in an icosahedral capsid made of 60 copies each containing four proteins: VP1, VP2, VP3, and VP4 ( 7). Seven serotypes ( 4) of this virus are observed including A, C, O, Asia 1, SAT1, SAT2 and SAT3 ( 5).įMD results in serious economic losses when outbreaks of the disease occur. The virus responsible for FMD is a picornavirus, a member of the genus Aphthovirus. Since FMD is highly infectious, it can be spread by polluted animals by aerosols, contact with contaminated farming equipment, vehicles, clothing, or feed, and through domestic and wild predators ( 3). FMD has severe implications for animal farming. This disease influences cloven-hoofed animals, such as domestic and wild bovines ( 2). Backgroundįoot-and-mouth disease (FMD) is an infectious and sometimes fatal viral disease ( 1). Specific Antibody S146 Antigen Foot-and-Mouth Disease Virus Serotype A Ion Exchange Chromatography 1. With respect to the results as well as the speed of ion exchange chromatography, this method is advised to purify antibodies.

Dot blot assay principle