重组酵母乙肝疫苗接种后鸡卵中抗乙肝IgY二级结构的圆二色谱分析

Circular dichroism chromatography of secondary structures of anti-HBV IgY in eggs from hens immunized with recombinant yeast HBV vaccine

目的用重组酵母乙肝疫苗免疫蛋鸡,探索抗乙肝IgY的产生和结构。方法产卵母鸡经3次免疫后收集所产卵,提取并纯化抗乙肝IgY,采用SDS-PAGE与反相高效液相层析(RP-HPLC)检测抗乙肝IgY,圆二色色谱(CD)测定抗乙肝IgY的二级结构。结果重组酵母乙肝疫苗表达的抗乙肝IgY相对分子质量(M r)为178 000。CD分析抗乙肝IgY二级结构α-螺旋、β-折叠、β-转角及无规则卷曲所含有的氨基酸残基数目分别为38、0、44和17。结论用重组酵母乙肝疫苗免疫蛋鸡能够表达抗乙肝IgY,α-螺旋、β-转角是抗乙肝IgY主要的二级结构。

Objective To study the generation and structure of anti-HBV IgY in eggs from hens immunized with recombinant yeast HBV vaccine. Methods From the hens immunized three times with the recombinant yeast HBV vaccine, the eggs were collected to extract anti-HBV IgY. After purified, the anti- HBV IgY was detected by sodium dodecyl sulfate- polyacrylamide gel electrophoresis （SDS-PAGE） and reverse phase high-performance liquid chromatography （RP-HPLC）, and its secondary structure was examined by circular dichroism （CD） chromatography. Results The relative molecular mass （Mr） of anti-HBV IgY expressed by the recombinant yeast HBV vaccine on hens was 178 000. CD chromatography showed that the （x-helix, β-fold, β-turn and random coil of anti-HBV IgY had 38, 0, 44 and 17 amino acids, respectively. Conclusion The anti-HBV IgY can be expressed in eggs from hens immunized with recombinant yeast HBV vaccine. The α-helix and β- turn are major secondary structures of anti-HBV IgY.