Simple Protocol for immunoglobulin G Purification from Camel “Camelus dromedarius” Serum

Abstract

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

Non‐enzymatic glycation and aggregation of camel immunoglobulins induce breast cancer cell proliferation

“…The sharp and single peak in each chromatogram illustrates the purity of isolated immunoglobulins. Previously, camel antibodies have been isolated and purified from serum 22 . Our simple 3‐step procedure led to the purification of (IgG1, IgG2, and IgG3) immunoglobulins from camel colostrum.…”

“…Previously, camel antibodies have been isolated and purified from serum. 22 Our simple 3-step procedure led to the purification of (IgG1, IgG2, and IgG3) immunoglobulins from camel colostrum. Similar strategies of immunoglobulin purification have been employed earlier, too.…”

“…Similar strategies of immunoglobulin purification have been employed earlier, too. 22,23 The protein concentrations in the IgG fractions were determined as 0.524 mg/mL (IgG1), 4.825 mg/mL (IgG2), and 2.12 mg/mL for IgG3. The relative molecular weights of the purified IgG subclasses, including conventional antibody and heavy-chain antibodies, were also estimated by SDS-PAGE under reducing conditions (Figure 2).…”

Development of R7BP inhibitors through cross-linking coupled mass spectrometry and integrated modeling

“…All purified antibodies were quantified by SDS–PAGE gel electrophoresis and stored in storage buffer at 4 °C until use. Alternatively, in order to remove excessive albumin from the serum and obtain higher antibody yields, total IgGs were isolated from llama serum using the caprylic acid purification method 53 by adjusting the serum pH to 5.5 and stirring with caprylic acid for 90 min, followed by centrifugation. The purified IgG was used for peptide-specific antibody purification as described above.…”

Comparative efficiency of ammonium sulfate alone and ammonium sulfate-octanoic acid combined methods in the purification potential of antibodies

“…Caprylic acid a short chain fatty acid that may precipitate the serum proteins other than IgG. This purification process gives a high yield of proteins, the drawback of caprylic acid is impurities that can be reduced through combined methods [11]. The present study was performed to evaluate the potential of ammonium sulfate and octanoic acid precipitation.…”