人乳铁蛋白多克隆抗体的制备、初步纯化及其应用

制备并初步纯化人乳铁蛋白的多克隆抗体，然后将抗体用于该蛋白的检测。分别对8周龄的昆明白小鼠和12周龄的新西兰白兔进行多次皮下免疫注射，采集血清并用硫酸铵沉淀法进行抗体的初步纯化。将初步纯化的抗体用于抗体夹心ELISA法检测转基因小鼠乳汁中人乳铁蛋白的表达量。经三次免疫后的新西兰白兔和昆明白小鼠血清中抗体的效价分别达到了1：48000和1：24000，并能在较高水平上维持两周以上，经初步纯化后抗体纯度得到很大提高。用抗体夹心EUSA法能够测出转基因小鼠乳汁中人乳铁蛋白含量。使用皮下注射的方法对昆明白小鼠和新西兰白兔进行免疫来制备多克隆抗体是可行的，硫酸铵沉淀法纯化后的抗体去除了大多数杂蛋白，可以满足测定的需要。

The Preparation, Preliminary Purification and Application of Human Lactoferrin Polycloned Antibody

In order to prepare and preliminarily purify the human lactoferrin polycloned antibody for detecting human lactoferrin in milk of transgenic mice, 8-week-old Kunmingbai mice and 12-week-old New Zealand white rabbits were immunized by hypodermic injection. Their sera were then collected and purified through precipitating using amine sulphate. Preliminarily purified polycloned antibody was applied to detect the human lactoferrin expression in milk of transgenic mice by using sandwich ELISA method. After the third immunity, the antibody titer in the serum of New Zealand white rabbits and Kunmingbai mice reached 1:48000 and 1:24000, respectively, and this level were maintained up to two weeks. The purity of the antibody was improved greatly after preliminary purification. Thus, the polycloned antibody can be prepared by immunizing Kunmingbai mice and New Zealand white rabbits by hypodermic injection. After the heterogenous proteins being cleaned out, the preliminarily purified antibody can be used in the detection of human lactoferrin.