Open Life Sci. 2017; 12: 143-155 was at 450 rpm. Overall, the results suggest that caprylic acid purification of camel serum IgG is more effective and safe than ammonium sulfate method in simplicity, purity, and lower non-IgG proteins in the final preparation with lower protein aggregates.Keywords: Camelus dromedarius; camel IgG purification; caprylic acid; temperature; stirring IntroductionCamels produce several immunoglobulin classes in their serum, colostrum and milk, including IgM, IgA and IgG (IgG1, IgG2 and IgG3) [1][2][3]. Camels are special animals in antibody production as they possess a unique type of antibody in their serum which lack light chains as well as the heavy chain constant domain "CH1", so-called heavychain antibodies [4]. IgG1 occurs in the classical structure form of antibodies, while IgG2 and IgG3 represent heavy-chain antibodies [4], and they form about 75% of the IgG in camel serum [5]. Heavy-chain antibodies in camels are significantly effective in antigen recognition; in spite of losing their light chains [6][7][8], also, they have lower molecular weight than conventional antibodies. In addition, these antibodies are less immunogenic than other mammalian antibodies [9], that means when they are injected into experimental models, it will be less likely to induce adverse reactions during the course of treatment [10]. The above pharmaceutical features recommend the camel antibodies as a substitute for other animals' therapeutic antibodies [11,12], or alternatively, it can be converted to a complete humanized antibody [13].Antibody purification from different biological fluids such as serum, ascites and milk is an important step in the production of antibodies used for diagnostic, therapeutic, and research applications [14]. There are different methods for purification and the first reported method for purifying camel IgG from serum, by , used protein A-Sepharose and protein G-Sepharose. More Abstract: The present study aimed to describe and standardize a simple and efficient protocol for purification of camel IgG from serum, which can be applied for Camilidae antibody production in research laboratories, the preindustrial stage. Camel serum IgG was separated with caprylic acid and ammonium sulfate, then the effect of four variables studied: caprylic acid concentration, pH, stirring time, and stirring intensity. Camel IgG prepared by standardized caprylic acid fractionation method for camel serum was compared with commercial anti-sera products. Camel IgG purification from undiluted sera using caprylic acid at concentration of 8% v/v gave the best results. Purification at different pH values using caprylic acid at 8% v/v revealed that pH 5.5 was optimal. Investigating purification at different stirring time intervals using 8% v/v caprylic acid at pH 5.5 demonstrated that stirring for 90 min gave the optimum results. Finally, studying purification at different stirring intensities using 8% v/v caprylic acid at pH 5.5 for 90 min, the best stirring intensity
A novel Ca'+ binding protein, named caligulin, was extracted from the heat-treated 100000 x g supernatant of bovine brain and purified to electrophoretic homogeneity. The apparent MI of caligulin determined on sodium dodecyl sulfate polyacrylamide gels was 24000. Analysis by gel filtration chromatography indicated an apparent M, of 33000, suggesting a monomeric protein. Amino acid composition data demonstrated the presence of 25% acidic residues, 12% basic residues and 10% leucine. In the presence of 1 mM MgClz and 0.15 M KCl, caligulin bound 1 mol Ca'+/mol protein with halfmaximal binding at about 0.2pM Ca'+.Cd' binding protein Brain
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