抗大肠杆菌O157鸡卵黄抗体的初步研究

Preparation of the anti-E.coli O157 antibody from egg yolk of the immunized chicken

目的以煮沸15min灭活后的大肠杆菌O157为抗原,分4次免疫产蛋鸡,获取高免卵黄。经水稀释法去脂、超滤、盐析等粗分离,DEAESephadexA-50离子交换层析纯化,得到产物IgY为电泳单点纯,且仍保持效价在1∶104以上。从分离纯化过程中的跟踪检测表明,其卵黄制备的IgY抗体产量多、效价高、特异性强,可进一步用于大肠杆菌感染的检测研究。

The antigen of E.coli O157 was heated with boiling for 15 minutes and used to immobilize chicken for 4 times, and the antibodies produced in the egg yolk were isolated and purified through the processes of ultrafiltration, salting-out and ion-exchange chromatography of DEAE-Sephadex A-50. The specific egg yolk immunoglobulin IgY produced was checked by SDS-PAGE and its titer was determined by ELISA. It was found that the titer of anti-E.coli O157 antibody produced by this method of immunization was above 1∶10 000. The egg yolk of the immunized chicken is a rich and inexpensive source of specific antibodies with large amount of specific and high titered antibodies produced, and can be used for the further studies of the E.coli infections.