McCormick D J, Hillman K, David C S
Department of Immunology, Mayo Clinic, Rochester, MN 55905.
J Immunol Methods. 1988 Oct 26;113(2):165-74. doi: 10.1016/0022-1759(88)90329-8.
synthetic peptides corresponding to residues 63-78 of the first domain of the beta chain of murine I-A/I-E class II antigens were used in the unconjugated rather than the traditional protein-conjugated form to immunize (129J X B6)F1 mice. The sequences made represented the four haplotypes; Ak beta, Ad beta, Abm-12 beta and Ed beta. These sequences were selected on the basis of computer algorithms used to predict surface accessibility and main-chain flexibility profiles, and by reported hypervariability and site-directed mutagenesis experiments of these regions. Factors such as the use of complete Freund's adjuvant, a continuous immunization regime, and the sex of the mice used were found to influence the amount of anti-peptide antibody produced when unconjugated peptide was used as the immunogen. Antibodies produced were shown by FACS analysis to react with I-A/I-E class II antigens expressed on the lymphocyte surface. These findings indicate that the use of unconjugated synthetic peptides representing sequences of proteins which are in close proximity to disulfide bonds may be an advantage over conventional methods of peptide conjugation to proteins.
对应于小鼠I-A/I-E II类抗原β链第一结构域63-78位残基的合成肽，以未偶联而非传统的蛋白偶联形式用于免疫（129J×B6）F1小鼠。所制备的序列代表四种单倍型：Akβ、Adβ、Abm-12β和Edβ。这些序列是根据用于预测表面可及性和主链柔韧性概况的计算机算法，以及这些区域已报道的高变异性和定点诱变实验来选择的。发现诸如使用完全弗氏佐剂、连续免疫方案以及所用小鼠的性别等因素，在以未偶联肽作为免疫原时会影响抗肽抗体的产生量。通过荧光激活细胞分选分析表明，所产生的抗体可与淋巴细胞表面表达的I-A/I-E II类抗原发生反应。这些发现表明，使用代表与二硫键紧密相邻的蛋白质序列的未偶联合成肽，可能比将肽与蛋白质偶联的传统方法更具优势。
