中文摘要：

虎纹捕鸟蛛（Ornithoctonus huwena）是中国最毒的蜘蛛之一.已有研究表明,其粗毒中含有丰富的低分子量（＜10kD）多肽活性成分.为分析这些成分,利用目标蛋白快速分离系统（ProteomeLab PF 2D）建立了一种新的二维液相色谱分离方法.该方法包括一维的基于蛋白质等电点（pI）的色谱聚焦分离和二维利用无孔硅胶反相柱的基于疏水性的高效液相色谱分离.得到的反相图谱通过仪器配套的ProteoVue软件转换成与凝胶电泳图像相似的pI/UV图,以更直观地显示多肽成分的数量、分布规律及相对丰度等.洗脱的多肽自动收集后用基质辅助激光解吸电离-飞行时间质谱进行分析.从一维分离的11个馏分（pH 4.53～8.59）中共检测到大约600个多肽条带.通过质谱分析测得130个多肽的精确分子量,同时通过De novo测序得到26种多肽（其中包括12种已知多肽）的部分序列信息,并利用这些序列信息对未知多肽进行了生物信息学分析.

英文摘要：

The bird spider Ornithoctonus huwena is one of the most venomous spiders in China. Previous studies have proved that its venom contains many biological peptides with molecular weight less than 10 kD. To analyze these components at the toxinome level, a novel two-dimensional liquid-phase separation method was developed on a protein purification system （ProteomeLab PF 2D）. In the first dimension, the peptides were fractionated on the basis of their pIs with a chromatofocusing column. In the second dimension, the peptides in each resulting fraction were separated on the basis of hydrophobicity with a noporous silica reversed-phase column. All the produced chromatograms were transformed into gel-like pI/UV maps by ProteoVue solfware attached to the instrument, which showed more intuitively the number, distribution and relative abundance of the peptides separated. Eluting peptides were automatically collected and analyzed by matrix-assisted laser desorption/ionization time of flight mass spectrometry. The pI/UV maps showed that about 600 peptides were detected, and these peptides came from 11 fractions （pH 4.53 ～ 8.59） obtained by chromatofocusing in the first dimension. Mass spectrometric analysis measured accurately the molecular weights of 130 peptides, and partial sequences of 26 peptides, including 12 known ones, were determined by de novo sequencing, which were employed to perform bioinformatics analysis of the unknown peptides.