Schmitz H E, Atassi H, Atassi M Z
Immunol Commun. 1983;12(2):161-75. doi: 10.3109/08820138309066866.
Two synthetic peptides corresponding to surface regions of sperm whale myoglobin that are not antigenic in the native molecule were used in their free form (i.e. not coupled to a carrier) to immunize separate groups of Balb/cByJ mice. The synthetic peptides corresponded to regions 1-6 and 121-127. Serum samples obtained from each group of mice contained antibodies that bound specifically to myoglobin and exclusively to the immunizing peptide. Monoclonal antibodies to each of the two surface regions were subsequently obtained by hybridizing Fa/O mouse myeloma cells with spleen cells derived from each group of mice. These monoclonal antibodies were IgM (kappa). They expressed the same isotype as the antigen specific serum antibodies produced by the mice whose spleen cells were used for hybridization. Solid phase radioimmunoassay studies also indicated that each monoclonal antibody, like the immune serum of the parent animals, bound specifically to myoglobin and exclusively to the synthetic peptide used as an immunogen. These results suggested that the hybridoma antibodies expressed submolecular binding specificities that were the result of peptide immunization rather than hybrid selection and that monoclonal antibodies with preselected submolecular binding specificities to non-antigenic surface regions in a protein molecule can be produced by the techniques of somatic cell hybridization when the corresponding free synthetic peptides are used as immunogens.
两种对应于抹香鲸肌红蛋白表面区域且在天然分子中无抗原性的合成肽，以游离形式（即未与载体偶联）用于免疫不同组的Balb/cByJ小鼠。这两种合成肽分别对应于1 - 6区和121 - 127区。从每组小鼠获得的血清样本中含有能特异性结合肌红蛋白且仅与免疫肽结合的抗体。随后，通过将Fa/O小鼠骨髓瘤细胞与每组小鼠的脾细胞杂交，获得了针对这两个表面区域的单克隆抗体。这些单克隆抗体为IgM（κ链）。它们与用于杂交的小鼠脾细胞所产生的抗原特异性血清抗体具有相同的同种型。固相放射免疫分析研究还表明，每种单克隆抗体与亲代动物的免疫血清一样，能特异性结合肌红蛋白且仅与用作免疫原的合成肽结合。这些结果表明，杂交瘤抗体表达的亚分子结合特异性是肽免疫的结果而非杂交选择的结果，并且当使用相应的游离合成肽作为免疫原时，通过体细胞杂交技术可以产生对蛋白质分子中非抗原性表面区域具有预选亚分子结合特异性的单克隆抗体。
