牙体牙髓牙周病学杂志
牙體牙髓牙週病學雜誌
아체아수아주병학잡지
Chinese Journal of Conservative Dentistry
2015年
8期
459-464,497
,共7页
董瑶%徐燕%马倩%孟明理%沈继龙%徐振山%宋礼华
董瑤%徐燕%馬倩%孟明理%瀋繼龍%徐振山%宋禮華
동요%서연%마천%맹명리%침계룡%서진산%송례화
牙龈卟啉单胞菌%免疫球蛋白Y%卵黄抗体
牙齦卟啉單胞菌%免疫毬蛋白Y%卵黃抗體
아간계람단포균%면역구단백Y%란황항체
porphyromonas gingivalis%immunoglobulin Y%egg yolk immunogolbuin
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( IgY)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IgY,BCA法测定蛋白质含量,ELLSA法检测纯化后IgY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-IgY 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 IgY抗体;SDS-PAGE分析,纯化的IgY有1条主要蛋白带,相对分子质量( Mr)为180 kD,纯度为65%; Western blotting分析, 该IgY抗体可识别抗原Pg. 每孔3. 125 μg Pg-IgY可有效抑制Pg的红细胞凝集活性;1 mg/mL Pg-IgY可有效抑制Pg生长.结论:本研究得到的IgY特异性强,产量和纯度高.
目的:製備特異性抗牙齦卟啉單胞菌( Pg)的鷄卵黃免疫毬蛋白( IgY)併檢測其生物學特性. 方法:厭氧培養牙齦卟啉單胞菌,將甲醛滅活的菌體經翅膀下淺層肌肉免疫150日齡囉曼母鷄3次,每次間隔10 d. 取鷄蛋用兩步硫痠銨沉澱法提取IgY,BCA法測定蛋白質含量,ELLSA法檢測純化後IgY的特異性和抗體效價變化,併進行 SDS-PAGE 和 Western blotting 分析. 觀察抗 Pg-IgY 對紅細胞凝集的抑製作用.結果:經3次免疫後,抗體滴度最高可達25 600,純化後每箇鷄蛋可得約70 mg IgY抗體;SDS-PAGE分析,純化的IgY有1條主要蛋白帶,相對分子質量( Mr)為180 kD,純度為65%; Western blotting分析, 該IgY抗體可識彆抗原Pg. 每孔3. 125 μg Pg-IgY可有效抑製Pg的紅細胞凝集活性;1 mg/mL Pg-IgY可有效抑製Pg生長.結論:本研究得到的IgY特異性彊,產量和純度高.
목적:제비특이성항아간계람단포균( Pg)적계란황면역구단백( IgY)병검측기생물학특성. 방법:염양배양아간계람단포균,장갑철멸활적균체경시방하천층기육면역150일령라만모계3차,매차간격10 d. 취계단용량보류산안침정법제취IgY,BCA법측정단백질함량,ELLSA법검측순화후IgY적특이성화항체효개변화,병진행 SDS-PAGE 화 Western blotting 분석. 관찰항 Pg-IgY 대홍세포응집적억제작용.결과:경3차면역후,항체적도최고가체25 600,순화후매개계단가득약70 mg IgY항체;SDS-PAGE분석,순화적IgY유1조주요단백대,상대분자질량( Mr)위180 kD,순도위65%; Western blotting분석, 해IgY항체가식별항원Pg. 매공3. 125 μg Pg-IgY가유효억제Pg적홍세포응집활성;1 mg/mL Pg-IgY가유효억제Pg생장.결론:본연구득도적IgY특이성강,산량화순도고.
AIM:To prepare egg yolk immunoglobulin ( IgY) against Porphyromonas gingivalis and to study its biological characteristics. METHODS:Ten 150 day-old laying hens were immunized with Porphyromonas gingi-valis and IgY was isolated from the egg yolk by the water dilution ( WD) method and 2 steps of ammonium sulfate puri-fication. The titers and specificity of the purified antibodies were assessed by ELLSA. The molecular weight were meas-ured by SDS-PAGE. The antigenicity and function of the IgY were demonstrated by western blotting and the hemag-glutination inhibition ( HI) test respectively. RESULTS:The yield of IgY was 70 mg per yolk with the purity of 65%. Its titer was 25 600. Pg-IgY could bound specifically to Porphyromonas gingivalis and could inhibit the hemagglutina-tion at 3. 125 μg/well. CONCLUSION:The production and purity of IgY are high, the IgY has strong specificity a-gainst T Porphyromonas gingivalis.