Green J M, Gu L, Ifkovits C, Kaumaya P T, Conrad S, Pierce S K
Department of Biochemistry, Molecular Biology, and Cell Biology Northwestern University, Evanston, Illinois 60208, USA.
Hybridoma. 1995 Aug;14(4):347-54. doi: 10.1089/hyb.1995.14.347.
The 70-kDa heat shock proteins (hsp70) are a highly conserved, abundant, and ubiquitous family of proteins expressed by all organisms from bacteria to humans. It is well established that hsp70 family members function as molecular chaperones and aid in the intracellular folding of newly synthesized or denatured proteins. Current evidence suggests an emerging role for hsp70 family members in immune responses and in clinically important responses to stress and tissue damage. Here we report the generation and characterization of several MAbs to hsp70 family members. Immune responses to this highly conserved family were induced in mice by immunization with synthetic peptides that contain regions of the mouse mitochondrial hsp70 coupled to a potent helper T cell epitope derived from tetanus toxoid. The resulting MAbs include ones specific for the human and mouse mitochondrial hsp70 and others that show cross-reactivity among the family members and recognize the mitochondrial hsp70, the endoplasmic reticulum resident hsp70, Bip/grp78, the constitutively expressed cytosolic hsp70, hsc70, and the heat-induced member, hsp70. Significantly, these MAbs are effective in Western blotting, in immunoprecipitation, and in immunofluorescence, and thus should find applications in the purification and detection of members of this important family.
70 kDa热休克蛋白（hsp70）是一个高度保守、丰富且普遍存在的蛋白质家族，从细菌到人类的所有生物体都能表达。众所周知，hsp70家族成员作为分子伴侣发挥作用，协助新合成或变性蛋白质的细胞内折叠。目前的证据表明，hsp70家族成员在免疫反应以及对应激和组织损伤的临床重要反应中发挥着新出现的作用。在此，我们报告了几种针对hsp70家族成员的单克隆抗体（MAb）的产生和特性。通过用包含小鼠线粒体hsp70区域并与源自破伤风类毒素的强效辅助性T细胞表位偶联的合成肽免疫小鼠，诱导对这个高度保守家族的免疫反应。产生的单克隆抗体包括对人和小鼠线粒体hsp70特异的抗体，以及其他在家族成员间显示交叉反应并识别线粒体hsp70、内质网驻留hsp70、Bip/grp78、组成性表达的胞质hsp70、hsc70和热诱导成员hsp70的抗体。重要的是，这些单克隆抗体在蛋白质印迹、免疫沉淀和免疫荧光中有效，因此应可用于该重要家族成员的纯化和检测。
