四种海水养殖鱼类血清免疫球蛋白的分离纯化及分子量测定

PURIFICATION AND CHARACTERISTICS OF SERUM IMMUNOGLOBULINS OF FOUR MAJOR CULTURED MARINE FISHES IN CHINA

利用ProteinA sepharose亲和柱层析一步法,首次分离纯化了4种海水养殖鱼类(3种暖水性鱼类:紫红笛鲷Lutjanusargentimaculatus、青石斑鱼Epinephelusawoara、尖吻鲈Latescal carifer和1种冷水性鱼类牙鲆Paralichthysolivaceus)血清中的免疫球蛋白(Ig)。3种暖水性鱼类Ig的分离效果较好,牙鲆较差。层析曲线的洗脱峰值说明牙鲆血清的Ig同紫红笛鲷、青石斑鱼、尖吻鲈3种暖水性鱼类血清的Ig在结构和功能上有明显的差异。通过SDS PAGE电泳测得其重链分子量分别为紫红笛鲷79.5kDa和75.0kDa、青石斑鱼77.2kDa、尖吻鲈81.8kDa和75.7kDa,牙鲆73.5kDa;轻链分子量分别为30.1kDa、31.1kDa、29.7kDa和15.0kDa。

Immunoglobulins of four major cultured marine fishes,mangrove red snapper Lutjanus argentimaculatus(Forskl),banded grouper Epinephelus awoara(Temminck et Schlegel),white seabass Lates calcarifer(Bloch) and Japanese flouder Paralichthys olivaceus(Temminck et Schlegel) were purified from the whole serum by affinity chromatography on protein Asepharose,a onestep purification method.The molecular weight of serum immunoglobulins was detected by SDSPAGE.The result showed that the molecular weights of the heavy chains were 79.5 kDa and 75.0 kDa for L.argentimaculatus,77.2 kDa for E.awoara,81.8 kDa and 75.7 kDa for L.calcarifer,and 73.5 kDa for P.olivaceus respectively; and those of the light chains were 30.1 kDa,31.1 kDa,29.7 kDa and 15.0 kDa respectively.