A Fast and Straightforward Method for the Purification of Anti- Immunoglobulin G (IgG) for Coombs Wright Assay

Journal of Medical Microbiology and Infectious Diseases Pub Date : 2020-10-01 DOI:10.29252/JOMMID.8.4.155

Z. Panahi, V. Kia, Mitra Moghiman, D. Doroud, R. Shokri, M. Paryan

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引用次数: 1

Abstract

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. 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. Finally, the Coombs Wright test was performed on samples from brucellosis patients to validate purified IgG antibody quality. Results: 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. Conclusion: 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. ,

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库姆斯·赖特试验中抗免疫球蛋白G (IgG)的一种快速、简便的纯化方法

纯化IgG。用SDS-PAGE和Bradford蛋白含量法评价纯化IgG的质量和浓度。每周给家兔注射四次不同量的蛋白质。然后取免疫小鼠血清，纯化总IgG。最后，对布鲁氏菌病患者样本进行Coombs Wright试验，以验证纯化的IgG抗体的质量。结果:电泳和Bradford实验结果表明，纯化蛋白具有相当高的纯度和数量。还原和非还原SDS-PAGE显示蛋白纯度高，蛋白产量为2.2 mg/L。库姆斯·赖特使用兔抗人血清进行的试验与现有的商用抗人免疫球蛋白的结果相当。结论:本方法可用于制备抗人球蛋白。该抗体也可用于临床诊断试验，如库姆斯赖特，交叉匹配和血型评估弱Rh或Du抗原。，

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来源期刊

Journal of Medical Microbiology and Infectious Diseases

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审稿时长

12 weeks