新玫瑰毒鲉毒素马血清抗体制备及抗毒活性研究

Preparation and antitoxic activity of horse anti-toxin from neoverrucotoxin

目的建立毒鲉毒素(俗称石头鱼毒素)马血清抗体的制备方法并研究其抗毒活性,为毒鲉毒素中毒提供治疗手段。方法利用排阻色谱制备毒鲉毒素,免疫马匹制备毒鲉毒素马抗,再制备不含FC的F(ab’)2并研究其对小鼠的抗毒活性。结果制备了新玫瑰毒鲉毒素(neoVTX)322.6 mg,纯度为88%,制备的马血清抗体原液中的蛋白含量为41 g/L,F(ab’)2的含量为87.05%,IgG含量为1.35%。4 LD50毒鲉粗毒素可被2倍质量的马抗完全中和;3 LD50neoVTX可被3倍质量的马抗完全中和。结论成功制备了毒鲉毒素及其马血清抗体,该抗体具有较高的抗毒活性,为毒鲉毒素中毒提供了治疗手段。

Objective To establish a preparation method of Synanceia(stonefish)horse antibody against neoverrucotoxin(neoVTX)and study its antitoxic activity so as to find a therapy for Synanceia toxin poisoning.Methods NeoVTX was prepared by size exclusion chromatography. F(ab’)2 antibody was prepared with horses immunized by Synanceia toxin,and its antitoxic activity was tested with mice.Results 322.6 mg of neoVTX was successfully prepared and the purity was 88%. The content of the horse antibody was 41 g/L.The purity of F(ab’)2 was 87.05% and that of IgG was 1.35%.The crude venom of stonefish(4LD50)could be completely neutralized by the horse antibody(1∶2,w/w),so could neoVTX(3LD50)(1∶3,w/w).Conclusion NeoVTX and its horse antibody are successfully prepared. The antibody has high antitoxic activity,which provides a method for the treatment of stone fish toxin poisoning.