Muller C P, Bühring H J, Becker G, Jung C C, Jung G, Tröger W, Saalmüller A, Wiesmuller K H, Bessler W G
Medizinische Universitätsklinik, Universität Tübingen, FRG.
Clin Exp Immunol. 1989 Dec;78(3):499-504.
Applying computer-assisted epitope prediction to the amino-acid sequence of the epidermal growth factor receptor (EGFR), the extracytoplasmic domain EGFR(516-529) was selected as a putative antigenic region. EGFR(516-529) was synthesized on a solid-phase matrix and N-terminally linked to the low mol. wt adjuvant tripalmitoyl-S-glyceryl-cysteinyl-serine (Pam3 Cys-Ser). The conjugation to this B cell and macrophage-activating lipopeptide considerably enhanced the immunogenicity of the EGFR peptide. Using the conjugate Pam3 Cys-Ser-EGFR(516-529), a peptide-specific monoclonal antibody was produced. By flow cytometry and immunoprecipitation the antibody was demonstrated to recognize EGFR on A431 cells, expressing large numbers of EGFR. With this novel approach synthetic immunogens can be prepared which could serve as thermostable synthetic vaccines with great potential in countries where a functional cold chain cannot be maintained.
将计算机辅助表位预测应用于表皮生长因子受体（EGFR）的氨基酸序列，胞外结构域EGFR(516 - 529)被选为假定的抗原区域。EGFR(516 - 529)在固相基质上合成，并在N端与低分子量佐剂三棕榈酰-S-甘油基-半胱氨酰-丝氨酸（Pam3 Cys-Ser）相连。与这种B细胞和巨噬细胞激活脂肽的偶联大大增强了EGFR肽的免疫原性。使用偶联物Pam3 Cys-Ser-EGFR(516 - 529)制备了一种肽特异性单克隆抗体。通过流式细胞术和免疫沉淀证明该抗体可识别表达大量EGFR的A431细胞上的EGFR。通过这种新方法可以制备合成免疫原，在无法维持功能性冷链的国家，这些合成免疫原可作为具有巨大潜力的热稳定合成疫苗。
