title: Study on camel IgG purification: a new approach to prepare Naja Naja Oxiana antivenom as passive immunization for therapy.
abstract: A combined process of ammonium sulfate precipitation (salting out) and ion-exchange chromatography on DEAE-Sepharose CL-6B was used to prepare camel antivenom (IgG) against Naja Naja Oxiana for therapy. In the ammonium sulfate precipitation, the best condition for fractionation of IgG from the other proteins in camel serum was 55% precipitate. The camel IgG presented as 2 bands with molecular masses of 250 and 100 kDa, the latter corresponding to heavy chain IgG, on 10% gel electrophoresis. A trace amount of non-IgG proteins was not isolated and remained in this precipitate. Therefore in order to effectively separate albumin and the other nonspecific proteins from the IgG, the 25% precipitate of ammonium sulfate precipitation of serum was subjected to DEAE-Sepharose CL-6B column chromatography. A peak of antibody (IgG) could be obtained by elution with sodium phosphate buffer. In this stage, 2 bands of molecular masses of 150 and 75 kDa were observed on 7% gel electrophoresis. A comparative study was performed between camel IgG and conventional horse F(ab) 2 antivenoms in term of potency (serum neutralization test and ELISA). Our results showed that the potency of camel antivenom was 4-fold higher than that of horse. It is suggested the combined ammonium sulfate precipitation and ion-exchange chromatography process effectively removed residual proteins in the final camel IgG preparation and can be a suitable method for large-scale refinement of therapeutic camel antivenoms.
authors.affiliation: Department of Venomous Animals and Antivenom; Razi Vaccine and Serum Research Institute; Karaj, Iran.
authors.forename: Sedigheh
authors.initials: S
authors.lastname: Khamehchian
authors.affiliation: Department of Venomous Animals and Antivenom; Razi Vaccine and Serum Research Institute; Karaj, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Zolfagharian
authors.affiliation: Department of Venomous Animals and Antivenom; Razi Vaccine and Serum Research Institute; Karaj, Iran.
authors.forename: Naser Mohammadpour
authors.initials: NM
authors.lastname: Dounighi
authors.affiliation: Department of Venomous Animals and Antivenom; Razi Vaccine and Serum Research Institute; Karaj, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Tebianian
authors.affiliation: Department of Biotechnology; Razi Vaccine and Serum Research Institute; Karaj, Iran.
authors.forename: Rasool
authors.initials: R
authors.lastname: Madani
mesh_terms: D000645:Ammonium Sulfate; D000818:Animals; D000997:Antivenins / Q000737:chemistry / Q000302:isolation & purification*; D002162:Camelus; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D017815:Elapidae / Q000276:immunology; D004586:Electrophoresis; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight
chemical_list: D000997:Antivenins; D007074:Immunoglobulin G; D000645:Ammonium Sulfate
keywords: IgG; ammonium sulfate; antivenom; camel; snake; therapy