大熊猫IgG Fc HRP酶标二抗的制备

Preparation of HRP enzyme-labeled second antibody against giant panda IgG Fc

为获得辣根过氧化物酶(HRP)标记的抗大熊猫Ig G抗体,本研究利用重组大肠杆菌表达技术对大熊猫Ig G Fc片段进行了表达,获得重组蛋白后进行了小鼠免疫,抗体特异性检测,并进行了多克隆抗体的HRP标记。结果显示,在37℃、IPTG浓度为0.5 mmol/L、诱导5 h的条件下,可获得分子质量约为88 ku的目的蛋白;将重组蛋白免疫小鼠后,免疫小鼠血清与重组蛋白反应效价可达1∶102400以上,且与大熊猫血清Ig G反应良好,经HRP标记试剂盒标记的鼠抗大熊猫Ig G效价可达1∶25600以上。上述结果表明,本研究成功制备了HRP标记的小鼠抗大熊猫Ig G二抗,为大熊猫疾病血清学检测方法的开发提供了支撑。

In order to obtain HRP labeled anti-giant panda Ig G antibody,the giant panda Ig G Fc fragment was expressed by recombinant E.coli expression technique.The recombinant protein was immunized in mice,the antibody specificity was detected,and the polyclonal antibody was labeled with HRP.The results showed that the target protein with molecular weight of about 88 ku could be obtained under the condition of 37℃,IPTG concentration of 0.5 mmol/L and induction for 5 h.After immunizing mice with the recombinant protein,the reaction titer of the immunized mouse serum with the recombinant protein was more than 102400 and reacted well with the giant panda serum Ig G.The titer of mouse anti-giant panda Ig G labeled by HRP labeling kit can reach more than 25600.The results showed that the HRP labeled mouse anti-giant panda Ig G secondary antibody was successfully prepared,which provided support for the development of serological detection methods for giant panda diseases.