Gearing A J, Thorpe R, Spitz L, Spitz M
J Immunol Methods. 1985 Feb 11;76(2):337-43. doi: 10.1016/0022-1759(85)90311-4.
We report here the use of 'single shot' intrasplenic injection of human IgM for immunization of mice to obtain splenocytes for use in the production of hybridomas secreting antibodies against human IgM. Fusion was performed 3 days after intrasplenic injection of 20 micrograms of myeloma IgM. IgM-specific antibodies were found in 12% of the fusion wells; only 1 well contained antibodies which cross-reacted with other immunoglobulin classes. Two monoclonal antibodies (McAbs) have been fully characterized as specific for different epitopes on Fc mu. These antibodies can be used to detect IgM on the surface of human B cells by immunofluorescence and in solution by solid-phase radiobinding assay or single radial immunodiffusion. Both McAbs can also detect IgM fragments by immunoblotting from non-reducing SDS-polyacrylamide gels.
我们在此报告通过脾脏内“单次注射”人IgM来免疫小鼠，以获取脾细胞用于制备分泌抗人IgM抗体的杂交瘤。在脾脏内注射20微克骨髓瘤IgM后3天进行细胞融合。在12%的融合孔中发现了IgM特异性抗体；只有1个孔中的抗体与其他免疫球蛋白类别发生交叉反应。两种单克隆抗体（McAbs）已被充分鉴定为对Fcμ上不同表位具有特异性。这些抗体可通过免疫荧光检测人B细胞表面的IgM，也可通过固相放射结合测定或单向免疫扩散在溶液中进行检测。两种单克隆抗体还可通过对非还原SDS-聚丙烯酰胺凝胶进行免疫印迹来检测IgM片段。
