文献类型：期刊文献

中文题名：红笛鲷仔鱼体内特异性母源抗体降解动力学研究

英文题名：DEGRADATION KINETICS OF LARVAL SPECIFIC MATERNAL ANTIBODY FROM CRIMSON SNAPPER(LUTJANUS ERYTHOPTERUS)

作者：彭志东[1];简纪常[1];吴灶和[1];鲁义善[1]

机构：[1]广东海洋大学水产学院,广东省水产经济动物病原生物学及流行病学重点实验室,湛江524025

年份：2009

卷号：33

期号：1

起止页码：34

中文期刊名：水生生物学报

外文期刊名：Acta Hydrobiologica Sinica

收录：CSTPCD、、CSCD2011_2012、北大核心2008、北大核心、CSCD

基金：国家自然科学基金(30471337);广东省自然科学基金(05106515);广东海洋大学项目资助

语种：中文

中文关键词：红笛鲷;仔鱼抗体;降解动力学;纯化

外文关键词：Lutjanus erythopterus ; Larval antibody ; Degradation kinetics ; Purification

中文摘要：应用山羊IgG免疫红笛鲷亲鱼,人工催产孵化出仔鱼后,用双抗体夹心法检测仔鱼特异性抗体的降解过程,发现抗体水平随时间变化呈下降趋势且可以持续7d,每尾仔鱼特异性抗体降解量约0.001μg,最低检测限为0.482μg/mL。采用饱和硫酸铵分步盐析与亲和层析等方法纯化红笛鲷仔鱼特异性抗体,结果显示,盐析仔鱼特异性抗体最理想的硫酸铵饱和度为40%—50%;亲和层析分离到两个明显独立的层析峰。SDS-PAGE证明纯化的抗体具有极高纯度,其L、H链分子量分别为29kD和77.5kD,抗体分子量约为852kD。该抗体不仅能够与山羊IgG产生特异性免疫反应,而且能与兔抗红笛鲷IgM抗血清产生免疫反应,因而认为红笛鲷母源抗体可以垂直转移至仔鱼。

外文摘要：The mature female crimson snapper （Lutjanus erythopterus） which was immunized with goat IgG propagated the larvae. Degradation kinetics of the larval specific antibody was analysed by using the double antibody sandwich ELISA. Purification and partial characterization of the specific antibody of larvae was studied. The purposes above were to elucidate the transfer mechanism of fish maternal antibody and increase the resistance diseases of larvae. The female fish were immunized by intraperitoneal （i. p. ） injection with the mixture of the goat IgG added equal volumes of FCA or FIA adjuvant four times interval two week during the period of egg development. After twenty days of final immunization, the fe- male fish were i. p. injected with the mixture of LRH-A3 ＋ HCG, then propagated the larvae. The results showed that the optimal concentration of the coating antigen, rabbit polyclonal antibody against crimson snapper IgM and horseradish peroxidase-labelled goat anti-rabbit IgG were 1. 563 μg/mL, 1 ： 800 and 1 ： 2000 respectively. The standard curve indicated that the detection limit was 0. 482μg/mL. The specific antibody of larvae degraded gradually and dropped to the lowest levels at 7 day after hatching. Meanwhile,ELISA showed that the specific antibody of larvae reacted with goat IgG and the serum of rabbit anti fish serum IgM ,which suggested that the specific antibody of larvae had some homology with the serum IgM. The specific antibody of larvae was purified from the larval homogenization using ammonium sulfate precipitation and affinity chromatography. The results showed that the larval antibody was salted out with 40% - 50% saturation of ammonium sul- fate. Two independent peaks were obtained with the affinity chromatography method. SDS-PAGE showed that other proteins were not found in the purified larval antibody. The molecular weights of light and heavy chain of the antibody were 29 kD and 77.5 kD respectively,and gross molecular weight was calculated to be 852 kD. It should play an important role in en- hancing resistance disease and viability of the larval fishes that the passively immunized mechanism through acquiring special maternal antibody via the yolk of the female fish immunized with antigen.