Welling G W, Geurts T, van Gorkum J, Damhof R A, Drijfhout J W, Bloemhoff W, Welling-Wester S
Laboratorium voor Medische Microbiologie, Rijksuniversiteit, Groningen, The Netherlands.
J Chromatogr. 1990 Jul 20;512:337-43. doi: 10.1016/s0021-9673(01)89500-5.
The possibility that a fragment of an antibody molecule may interact with a protein antigen was tested by studying the binding properties of a thirteen-residue synthetic peptide with an amino acid sequence similar to part of a hypervariable segment of a monoclonal antibody directed against lysozyme. Affinity adsorbents were prepared with this peptide and with non-related peptides as ligand. Non-specific interactions could be abolished by washing the column with 0.05 M sodium thiocyanate in 20 mM tris-HCl (pH 7.4). Lysozyme was only bound to the antilysozyme adsorbent and could be eluted with 1 M sodium thiocyanate. The results show that immunoaffinity chromatography with synthetic peptide ligands which mimic the antigen-binding site may be a useful tool in the selective purification of proteins.
通过研究一种十三肽的结合特性来测试抗体分子片段与蛋白质抗原相互作用的可能性,该十三肽的氨基酸序列与针对溶菌酶的单克隆抗体高变区的一部分相似。用该肽和无关肽作为配体制备亲和吸附剂。用0.05M硫氰酸钠在20mM三羟甲基氨基甲烷盐酸盐(pH 7.4)中洗涤柱子可消除非特异性相互作用。溶菌酶仅与抗溶菌酶吸附剂结合,可用1M硫氰酸钠洗脱。结果表明,用模拟抗原结合位点的合成肽配体进行免疫亲和层析可能是蛋白质选择性纯化的一种有用工具。