Kubiak T, Whitney D B, Merrifield R B
Rockefeller University, New York, New York 10021.
Biochemistry. 1987 Dec 1;26(24):7849-55. doi: 10.1021/bi00398a047.
A 53-residue peptide corresponding to the variable region 16-68 of the heavy chain of phosphocholine binding mouse myeloma M603 protein was synthesized by a solid-phase fragment strategy. The homogeneity of the VH(16-68) peptide was confirmed by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, amino acid analysis, and mass spectrometry. Synthetic VH(16-68) associated with the M603 light chain, and about 27% of the recombination mixture bound to phosphocholine immobilized on Sepharose as compared to a 28% binding yield obtained for the recombined natural light and heavy chains under the same conditions. The binding yield for the recombinant of the light chain with previously prepared VH(27-68) fragment was about 11%. These semisynthetic antibodies VH(27-68) and VH(16-68) light chain recombinants are forerunners of structural variants designed to study the antigen binding pocket of the M603 immunoglobulin.
通过固相片段策略合成了一种53个氨基酸残基的肽，其对应于磷酸胆碱结合小鼠骨髓瘤M603蛋白重链的可变区16 - 68。通过高效液相色谱、十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳、氨基酸分析和质谱法确认了VH(16 - 68)肽的纯度。合成的VH(16 - 68)与M603轻链结合，在相同条件下，重组混合物中约27%与固定在琼脂糖上的磷酸胆碱结合，而重组天然轻链和重链的结合产率为28%。轻链与先前制备的VH(27 - 68)片段重组的结合产率约为11%。这些半合成抗体VH(27 - 68)和VH(16 - 68)轻链重组体是为研究M603免疫球蛋白的抗原结合口袋而设计的结构变体的先驱。
