建立分泌蒙古沙鼠（Meriones unguiculatus）免疫球蛋白的沙鼠-鼠杂交瘤

Establishment of gerbil-mouse heterohybridoma secreting immunoglobulin of Mongolian gerbil (Meriones unguiculatus) establishment of gerbil-mouse heterohybridoma.

机构信息

Department of Animal Science and Resources, College of Bioresource Sciences, Nihon University, Kameino 1866, Fujisawa 252-0880, Japan.

出版信息

J Immunol Methods. 2011 Oct 28;373(1-2):174-80. doi: 10.1016/j.jim.2011.08.015. Epub 2011 Aug 31.

DOI:10.1016/j.jim.2011.08.015

PMID:21907207

Abstract

Hybridomas, generated by fusing myeloma cells with B lymphocytes, secrete monoclonal antibodies (mAbs) specific for an antigen. To establish hybridomas that secrete Mongolian gerbil (Meriones unguiculatus) mAbs, we immunized gerbils with keyhole limpet hemocyanin (KLH) and fused splenocytes from them with a non-secreting mouse myeloma cell line (P3-X63-Ag8.653). We obtained hybridomas that secreted immunoglobulin (Ig) against KLH. These hybridomas had more chromosomes than either parent and retained several gerbil chromosomes. Therefore, these cells were heterohybridomas with both gerbil and mouse chromosomes. They expressed gerbil Ig heavy-chain constant (IGHC) region mRNA, but not mouse gamma (γ) 1 IGHC. SDS-PAGE and Western blot analyses indicated that the cells secreted whole Ig molecules of gerbil origin. Moreover, the cells continued secreting Ig for several months.

摘要

杂交瘤由骨髓瘤细胞与 B 淋巴细胞融合而成，可分泌针对特定抗原的单克隆抗体（mAbs）。为了建立分泌蒙古沙鼠（Meriones unguiculatus）mAbs 的杂交瘤，我们用血蓝蛋白（KLH）免疫沙鼠，并将来自它们的脾细胞与不分泌的小鼠骨髓瘤细胞系（P3-X63-Ag8.653）融合。我们获得了分泌针对 KLH 的免疫球蛋白（Ig）的杂交瘤。这些杂交瘤比其亲本具有更多的染色体，并且保留了几条沙鼠染色体。因此，这些细胞是具有沙鼠和小鼠染色体的异源杂交瘤。它们表达沙鼠 Ig 重链恒定区（IGHC）mRNA，但不表达小鼠 γ1 IGHC。SDS-PAGE 和 Western blot 分析表明，这些细胞分泌源自沙鼠的完整 Ig 分子。此外，这些细胞可以连续数月持续分泌 Ig。