公开(公告)号：KR100353473B1

公开(公告)日：2002-09-26

申请号：KR1019990046797

申请日：1999-10-27

Applicant: 대한민국(농촌진흥청장)

Inventor： 채현석 ,  김동운 ,  안종남 ,  조성근 ,  심정석 ,  김용곤

IPC: C12P21/00

Abstract: 본발명은최근육가공품에서식중독을일으켜문제가되고있는 E. coli O157: H7균을효과적으로방지하기위하여기존의방법인혈액을이용하지않고산란계의면역생리를이용하여 E. coli O157: H7 균을액체배지를이용하여배양하고균을죽인다음원심분리를통해균을회수한후 균질화시켜항원으로이용하였으며이 항원을산란계에주사하여 E. coli O157: H7균의항체를다량함유한계란을생산하고, 이계란에서난황내에함유된항체를분리하며분리된액상의항체를바로이용하거나열풍건조시켜사용하거나상기에서만들어진 E. coli O157 항체를함유한전란액에혼합시켜강화된동결액란을제조하여보존성이편리하고필요에따라해동하여햄버거용패티와같은육가공품의원료로이용할수 있도록한 E. coli O157: H7균의증식을억제시키는항체를제조하는방법에관한것이다.

公开(公告)号：KR1020010038702A

公开(公告)日：2001-05-15

申请号：KR1019990046797

申请日：1999-10-27

Applicant: 대한민국(농촌진흥청장)

Inventor： 채현석 ,  김동운 ,  안종남 ,  조성근 ,  심정석 ,  김용곤

IPC: C12P21/00

Abstract: PURPOSE: Provided is a method for manufacturing antibody of E. coli O157:H7 in a high yield by using eggs which can be more easily obtained than blood of rabbits to prevent food poisoning beforehand and help farmers increase their income. CONSTITUTION: A method for manufacturing antibody of E. coli O157:H7 is characterized by the next steps of: i) cultivating E. coli O157:H7 in brain heart infusion broth, killing the bacteria in water of 90-100 deg.C for 5-10 minutes, and centrifuging the broth to obtain only the bacteria; ii) using a sonicator to destroy E. coli, and washing and drying the destroyed bacteria; iii) mixing 1-5 mg of dried bacteria, 0.5 ml of PBS buffer solution (pH 7.2), and 0.5 ml of Freund's complete adjuvant to make an antigen; iv) inserting the antigen into hens about to lay eggs and repeating the process two weeks later; v) selecting eggs 40- 60 days after the insertion; vi) separating immunoglobin Y from the eggs and storing the antibodies at 4 deg.C for 6-12 hours; vii) centrifuging the antibodies at 7,000 rpm for 20 minutes to obtain supernatant and drying the antibodies; and viii) mixing dried antibodies with the eggs of step (v) and freezing the eggs to increase the amount of antibodies.

Abstract translation: 目的：提供通过使用可以比兔血液更容易获得的蛋，以预防食物中毒并帮助农民增加收入，以高产率制造大肠杆菌O157：H7的抗体的方法。 构成：制备大肠杆菌O157：H7抗体的方法的特征在于以下步骤：i）在脑心脏输液肉汤中培养大肠杆菌O157：H7，将90-100℃的水中的细菌杀死 5-10分钟，离心肉汤只得到细菌; ii）使用超声波破坏大肠杆菌，洗涤和干燥被破坏的细菌; iii）混合1-5mg干燥细菌，0.5ml PBS缓冲溶液（pH7.2）和0.5ml弗氏完全佐剂以制备抗原; iv）两周后，将抗原插入母鸡中以产卵，重复该过程; v）插入后40-60天选择鸡蛋; vi）从蛋中分离免疫球蛋白Y并将抗体在4℃下储存6-12小时; vii）以7,000rpm离心抗体20分钟以获得上清液并干燥抗体; 和viii）将干燥的抗体与步骤（v）的卵混合并冷冻卵以增加抗体的量。