en Diagnostic/screening methods and protocols Diagnostic/screening methods and protocols Original article Original article <table border="0" cellpadding="0" cellspacing="0" style="width:101.14%;" width="101%"> <tbody> <tr> <td style="width:69.24%;height:24px;"><strong>Introduction:</strong> Immunoglobulin G is the most abundant immunoglobulin in human blood, comprising approximately 75% of serum antibodies. Human IgG is a glycoprotein and can be an antigen for other mammals. Antibody-sensitized red blood cells (RBCs) and complement components can react with anti-human globulins resulting in their agglutination. This study aimed to prepare an antiserum against human IgG for deploying in Coombs Wright and cross-match for rare blood groups. <strong>Methods:</strong> After isolation of serum from healthy blood donors, serum proteins were precipitated using ammonium sulfate. Consequently, tangential flow filtration and ion-exchange chromatography were applied to purify IgG. SDS-PAGE and Bradford protein content assay was conducted to evaluate the quality and the concentration of the purified IgG.&nbsp; Rabbits were weekly injected with different amounts of the protein four times. Then, sera were obtained from the immunized mice, and total IgG was purified.&nbsp; Finally, the Coombs Wright test was performed on samples from brucellosis patients to validate purified IgG antibody quality. <strong>Results:</strong> Electrophoresis and Bradford assay results showed that the purified protein had considerable high purity and quantity. Protein bands of reducing and the non-reducing SDS-PAGE showed high purity of the protein along with a protein yield of 2.2 mg/L. Coombs Wright tests using the rabbit anti-human serum had a comparable result with available commercial anti-human immunoglobulin. <strong>Conclusion:</strong> The results indicated that our method for the purification of IgG was suitable for anti-human globulin preparation. This antibody can also be used in clinical diagnostic tests such as Coombs Wright, cross-match, and blood types evaluation with weak Rh or Du antigens.&nbsp;</td> </tr> </tbody> </table> Anti-human globulin, Immunoglobulins, Coombs Wright 155 160 http://jommid.pasteur.ac.ir/browse.php?a_code=A-10-307-1&slc_lang=en&sid=1 Zahra Panahi zahrapanahi866@yahoo.com 0000-0002-4020-971X No Danesh Alborz University, Qazvin, Iran Vahid Kia vahidkiaa@gmail.com 0000-0001-8617-1465 No Department of medical biotechnology, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran Mitra Moghiman mitramoghiman@gmail.com 0000-0002-7788-142X No Danesh Alborz University, Qazvin, Iran Delaram Doroud d_doroud@yahoo.com 0000-0003-4167-9921 No Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran Rahman Shokri shokrei@gmail.com 0000-0002-1996-5799 No Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran mahdi paryan mparyan@gmail.com 0000-0002-2592-3528 Yes Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran