Purification of vitellin from the ovary of Chinese mitten-handed crab (Eriocheir sinensis) and development of an antivitellin ELISA.
Vitellin was purified from ovaries of mature female Chinese mitten-handed crab (Eriocheir sinensis) using gel filtration chromatography. Analysis by native PAGE showed the vitellin had a native molecular mass of 520 kDa, while denaturing SDS-PAGE revealed two subunits of 97 and 74 kDa. Purified vitellin was used to raise polyclonal antisera, with which an enzyme-linked immunosorbent assay (ELISA) was developed. The ELISA was sensitive and could effectively detect vitellin in the range of 7.8-500 ng. Furthermore, vitellin levels in various developmental stages of oogenesis were measured with the ELISA assay. The results indicated that levels of vitellin increased significantly from 0.22 mg/ovary at Stage II to 360.31 mg/ovary at Stage IV.
Chen
Liqiao
L
Department of Biology, East China Normal University, Shanghai 200062, PR China. lqchen@bio.ecnu.edu.cn
Jiang
Hongbo
H
Zhou
Zhongliang
Z
Li
Kang
K
Li
Kai
K
Deng
Glenn Y
GY
Liu
Zhanjiang
Z
D000818:Animals; D003386:Brachyura / Q000378:metabolism*; D002681:China; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008297:Male; D010053:Ovary / Q000378:metabolism*; D011817:Rabbits; D047048:Vitellins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*
D047048:Vitellins