【摘要】 目的:制备特异性抗牙龈卟啉单胞菌(Pg)的鸡卵黄免疫球蛋白(Ig Y)并检测其生物学特性。方法:厌氧培养牙龈卟啉单胞菌,将甲醛灭活的菌体经翅膀下浅层肌肉免疫150日龄罗曼母鸡3次,每次间隔10 d。取鸡蛋用两步硫酸铵沉淀法提取Ig Y,BCA法测定蛋白质含量,ELLSA法检测纯化后Ig Y的特异性和抗体效价变化,并进行SDS-PAGE和Western blotting分析。观察抗Pg-Ig Y对红细胞凝集的抑制作用。结果:经3次免疫后,抗体滴度最高可达25 600,纯化后每个鸡蛋可得约70 mg Ig Y抗体;SDS-PAGE分析,纯化的Ig Y有1条主要蛋白带,相对分子质量(Mr)为180 k D,纯度为65%;Western blotting分析,该Ig Y抗体可识别抗原Pg。每孔3.125μg Pg-Ig Y可有效抑制Pg的红细胞凝集活性;1 mg/m L Pg-Ig Y可有效抑制Pg生长。结论:本研究得到的Ig Y特异性强,产量和纯度高。更多还原

【Abstract】 AIM: To prepare egg yolk immunoglobulin( Ig Y) against Porphyromonas gingivalis and to study its biological characteristics. METHODS: Ten 150 day- old laying hens were immunized with Porphyromonas gingivalis and Ig Y was isolated from the egg yolk by the water dilution( WD) method and 2 steps of ammonium sulfate purification. The titers and specificity of the purified antibodies were assessed by ELLSA. The molecular weight were measured by SDS- PAGE. The antigenicity and function of the Ig Y were demonstrated by western blotting and the hemagglutination inhibition( HI) test respectively. RESULTS: The yield of Ig Y was 70 mg per yolk with the purity of 65%.Its titer was 25 600. Pg- Ig Y could bound specifically to Porphyromonas gingivalis and could inhibit the hemagglutination at 3. 125 μg / well. CONCLUSION: The production and purity of Ig Y are high,the Ig Y has strong specificity against T Porphyromonas gingivalis.更多还原