抗耐甲氧西林金黄色葡萄球菌(MRSA)卵黄免疫球蛋白(IgY)的制备与性质研究

目的观察鸡卵黄中抗耐甲氧西林金黄色葡萄球菌(methicillin-resistant staphylococcus aureus,MRSA)抗体的含量、纯度、来源及稳定性。方法用耐甲氧西林金黄色葡萄球菌(MRSA)作为抗原免疫Leghorn鸡;通过改良水稀释法提取卵黄中的IgY;双紫外光波长测定抗体含量,SDS-PAGE电泳检测抗体纯度;Western blot免疫印迹法测定该抗体来源;ELISA检测IgY对温度、酸碱度的稳定性。结果抗体含量13.3 mg/mL蛋黄液,抗体纯度达96%;免疫印迹证明IgY与鸡血清中的IgG具有相同的分子量和抗原性;IgY具有良好的热稳定性,对酸碱具有一定的耐受力。结论改良水稀释法能得到高产量、高纯度的特异性IgY,而且有良好的生物学活性。

The preparation and character investigation of the immunoglobulin of yolk (IgY) produced by methicillin-resistant staphylococcus aureus (MRSA)

Objective To observe the yield,purity,source,and stability of the methicillin-resistant staphylococcus aureus(MRSA).Methods Taken the methicillin-resistant staphylococcus aureus(MRSA) as antigen immuned Leghorn fowl,IgY was extracted with improved water dilution method.Antibody's content was determined with double beam UV/VIS spectrophotometer,and antibody purity was detected with SDS-PAGE electrophoresis.Antibody's source was determined by Western blot.IgY's constancy to temperature and acidity or alkalinity was determined with ELISA.Results Antibody contained yolk liquid at 13.3 mg/mL;purity of antibody: 96%;immunoblotting showed that IgY and IgG in chicken serum had the same molecular weight and antigenicity,IgY had favorable heat stability and tolerance to acid and base.Conclusion High yield of IgY that had highly purified specificity and favorable biologic activity can be prepared with improved water dilution method.