Haba S, Nisonoff A
Department of Biology, Brandeis University, Waltham, MA 02254.
J Immunol Methods. 1987 Dec 24;105(2):193-9. doi: 10.1016/0022-1759(87)90266-3.
Methods are described for the production of syngeneic mouse anti-IgE monoclonal antibodies (mAb). Hybridomas were prepared by using spleen cells from mice immunized with a conjugate of keyhole limpet hemocyanin with monoclonal IgE. Serum titers varied from approximately 40 to 1000 micrograms/ml. The anti-IgE mAb were isolated by affinity chromatography on columns containing immobilized monoclonal IgE. The mAb are specific for isotypic determinants of IgE and do not react with other immunoglobulin isotypes. One of the mAb, which has a high affinity for IgE (Ka = 4.7 X 10(8) M-1), should be useful for studies of regulation of IgE. The applicability of the mAb to quantitative assays for IgE was demonstrated.
描述了产生同基因小鼠抗IgE单克隆抗体(mAb)的方法。通过使用用钥孔戚血蓝蛋白与单克隆IgE的缀合物免疫的小鼠的脾细胞制备杂交瘤。血清滴度约为40至1000微克/毫升。通过在含有固定化单克隆IgE的柱上进行亲和层析分离抗IgE mAb。这些mAb对IgE的同种型决定簇具有特异性,并且不与其他免疫球蛋白同种型反应。其中一种对IgE具有高亲和力(Ka = 4.7×10(8)M-1)的mAb,应用于IgE调节的研究。证明了该mAb在IgE定量测定中的适用性。