Cao Y-L, Guo G-N, Zhu G-Y, Tian Z, Gou Y-J, Chen C, Liu M-H
Department of Emergency, Southwest Hospital, Third Military Medical University, Chongqing, China.
Eur Rev Med Pharmacol Sci. 2016;20(4):781-7.
With the help of bioinformatics analysis, we wished to develop a novel antivenom against the Deinagkistrodon (D.) acutus snake venom using B-cell linear epitopes of three primary toxins (serine protease, metalloprotease, and phospholipase A2).
cDNA sequences of three toxins of D. acutus venom were retrieved from the NCBI database. B-cell linear epitopes were predicted using DNAStar and the website server software provided by IEDB. Then, the sequences of the predicted epitopes were artificially synthesized and inserted into the vector pET-32a-c(+). Recombinant antigen peptide was expressed and purified. BALB/c mice were immunized with the recombinant antigen peptide. The immunoprotective effect of this novel antivenom was measured by neutralization of venom haemorrhagic activity.
Six epitopes were obtained by bioinformatics analysis. ELISA analysis showed that antibody titre was >8,000 against snake venom and >64,000 against the recombinant peptide. Neutralization assays confirmed that the developed antivenom could effectively reduce the haemorrhagic activity of snake venom.
Six B-cell linear epitopes of D. acutus snake venom were predicted by bioinformatics analysis and successfully utilized to produce a novel antivenom.
借助生物信息学分析，我们希望利用三种主要毒素（丝氨酸蛋白酶、金属蛋白酶和磷脂酶A2）的B细胞线性表位开发一种针对尖吻蝮蛇毒的新型抗蛇毒血清。
从NCBI数据库检索尖吻蝮蛇毒三种毒素的cDNA序列。使用DNAStar和IEDB提供的网站服务器软件预测B细胞线性表位。然后，人工合成预测表位的序列并将其插入载体pET-32a-c(+)。表达并纯化重组抗原肽。用重组抗原肽免疫BALB/c小鼠。通过中和毒液出血活性来测定这种新型抗蛇毒血清的免疫保护效果。
通过生物信息学分析获得了六个表位。ELISA分析表明，抗蛇毒血清的抗体效价>8000，抗重组肽的抗体效价>64000。中和试验证实，所开发的抗蛇毒血清可有效降低蛇毒的出血活性。
通过生物信息学分析预测了尖吻蝮蛇毒的六个B细胞线性表位，并成功用于生产新型抗蛇毒血清。
