TRYPTIC DIGESTION OF INTERFERON COMPONENTS

从Wellferon中分离纯化的22种干扰素组分 将用胰蛋白酶TPCK消化。 全部消化 将在37摄氏度下进行16至18小时。 的 然后将消化的材料通过C-18 HPLC柱以分离 个别的碎片。 这将使我们能够产生胰蛋白酶 每个组分的消化图谱（色谱图）。 每个人的碎片 然后收集组分并测定蛋白质序列， 氨基酸分析 到目前为止，我们只能部分地 表征组分。 通过对消化片段进行测序，我们 应该能够确定每个组分的完整蛋白质序列。 氨基酸分析将提供每种化合物的组成评价。 成分 如果我们要开发出一种新的分子， 疏水性和抗原性谱以及确定 组件的二级结构。

Twenty two interferon components which have been purified from Wellferon in our laboratory will be digested using trypsin TPCK. All digestion will be carried out at 37 degrees C for a period of 16 to 18 hours. The digested material will then be passed over a C-18 HPLC column to separate the individual fragments. This will enable us to generate a tryptic digest map (chromatogram) for each component. The fragments from each component will then be collected and assayed for protein sequence and amino acid analysis. To date, we have only been able to partially characterize the components. By sequencing the digestion fragments, we should be able to determine the full protein sequence for each component. The amino acid analysis will provide a compositional evaluation of each component. Characterization of the entire molecule is important if we are to develop hydrophobicity and antigenicity profiles as well as determining the secondary structure of the component.