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. Keywords:IgG, ammonium sulfate, camel, antivenom, snake, therapy == Intro == Antivenoms have been used successfully for more than a century and up to right now constitute the only effective treatment for snakebites and envenomations by other poisonous animals.1,2Currently, a great variety of antivenoms are produced in many laboratories from your serum of animals, mostly horses and sheep, that are immunized with single or multiple venoms. observed on 7% gel electrophoresis. A comparative study was performed between camel IgG and standard horse F(abdominal)2antivenoms in term of potency (serum Amyloid b-Protein (1-15) neutralization test and ELISA). Our results showed the potency of camel antivenom was 4-collapse higher than that of horse. It is suggested the combined ammonium sulfate precipitation and ion-exchange chromatography process effectively eliminated residual proteins in the final camel IgG preparation and can be a suitable method for large-scale refinement of restorative camel antivenoms. Keywords:IgG, ammonium sulfate, camel, antivenom, snake, therapy == Intro == Antivenoms have been used successfully for more than a century and up to right now constitute the only effective treatment for snakebites and envenomations by additional poisonous animals.1,2Currently, a great variety of antivenoms are produced in many laboratories from your serum of animals, mostly horses and sheep, that are immunized with single or multiple venoms. The 3 main forms of antivenom preparations are IgG, F(ab)2,and Fab.3,4All the preparations are ineffective in the local venom effects because of their failure to penetrate into the blood/cells barrier.5Moreover, early and delayed adverse reactions are often associated with parenteral antivenom administration6and their incidence, depending upon the administrated product, is highly different.7,8Some causes of adverse reactions to antivenoms are immunogenicity of antivenom proteins3,8,9and complement activation due to the presence of Fc in the whole IgG preparations or protein aggregates in 3 forms of antivenoms.3,10,11Immunogenicity of antivenom proteins which would elicit an anti-IgG antibody response associated with delayed reactions, i.e., serum sickness.3,8,9 Studies on camelid immunoglobulin symbolize an attractive alternative in developing an antibody-based approach to the treatment of envenoming. Unique physicochemical properties of camelied IgG present intriguing options to improving the clinical performance of antivenom treatment.12Over 50% of camelid immunoglobulin lack light chains13these unique Ab isotypes interact with the antigen by virtue of only one single V domain. Solitary website Abs isolated from camel H-chain Abs are known as VHH or nanobody and are adaptable to screening techniques such as phage display which provides the fast and easy isolation of specific Ab.14,15Therefore, VHH is considerably smaller than Fab fragment (produced by papain effect) of conventional IgG.5Immunogenicity of camel IgG is low and less likely to activate the match cascade than ovine or equine IgG16; therefore, the anaphilactoid and serum sickness adverse effects in the individuals treated with camelid IgG antivenom would be less. Amyloid b-Protein (1-15) These effects are often associated with the current antivenom treatment.16The unusual thermostability of camelid IgG17can be exploited to prepare antivenom that remains efficacious after maintenance at room temperature.16It is supposed that the major clinical advance will be achieved by IgG from venom-immunized camelids to provide antivenom which is capable of treating snake envenoming. Amyloid b-Protein (1-15) In addition, this animal is definitely facile in terms of handling, immunization, and bleeding like horsesthe yield of blood is similar to that of the horse.16 Systems for snake antivenom MYO9B production vary according to the production laboratory, even in the same country. Classically, the protocols that yield the F(ab)2fragment based on pepsin digestion followed by ammonium sulfate fractionation have been used worldwide.1This method routinely is applied for the production of antivenom in horse against the Naja Naja Oxiana snake venom in Razi Institute, Iran. The aim of this work was 2-folds, (1) to adapt the methodology used for ammonium sulfate purification (without pepsin) of horse F(ab)2to camel IgG, in order to develop an alternative method for antivenom preparation for therapy, and (2) to compare the proposed method with standard one in terms of potency. == Results == == Antibody production == With this work, the camel immunized with different concentrations of Naja Naja Oxiana snake venom. The antivenom antibodies appeared in the serum during the third time of immunization (1st bleeding) and reached the hyperimmune level in the eighth immunization step (fourth bleeding). This immunity was exposed by a double immunodiffusion method (data not demonstrated). The regularly horse F(ab)2antivenom used as a positive control. The hyper- immunity of both animals (camel and horse) was necessary for processing in the next methods and comparative studies. == Ammonium sulfate purification of IgG == The procedure which was used for the purification of horse F(ab)2based on.