Sithigorngul Paisarn, Nanda Jennifer Cho, Stretton Antony O W
Department of Zoology, University of Wisconsin-Madison, 1117 W. Johnson Street, Madison, WI 53706, USA.
Peptides. 2003 Jul;24(7):1025-33. doi: 10.1016/s0196-9781(03)00184-0.
Monoclonal antibody G15-6A was generated by immunizing mice with Ascaris head extracts. It recognizes an antigen present in a single neuron, with a cell body in the dorsal rectal ganglion, that projects along the ventral cord to the nerve ring. Ascaris extracts were fractionated by HPLC and ammonium sulfate precipitation, and fractions assayed by dot-blotting with antibody G15-6A. A single immunoreactive polypeptide was purified; mass spectrometry showed a molecular weight of 11,542 Da. Partial N-terminal sequencing, followed by cloning of the transcript encoding the peptide, revealed a predicted peptide product comprising 109 amino acids, and a molecular mass of 11,863 Da. The N-terminus of the predicted peptide includes four more amino acids than are found in the isolated product.
单克隆抗体G15-6A是通过用蛔虫头部提取物免疫小鼠产生的。它识别存在于单个神经元中的一种抗原，该神经元的细胞体位于直肠背侧神经节，其沿着腹索延伸至神经环。蛔虫提取物通过高效液相色谱法（HPLC）和硫酸铵沉淀进行分级分离，并用抗体G15-6A通过斑点印迹法对各组分进行检测。一种单一的免疫反应性多肽被纯化；质谱分析显示其分子量为11542道尔顿。通过部分N端测序，随后克隆编码该肽的转录本，揭示了一个预测的肽产物，其包含109个氨基酸，分子量为11863道尔顿。预测肽的N端比分离产物中发现的氨基酸多四个。
