中文摘要：

Gp96， HSP90 家庭的一个成员，是有各种各样的最新发现的函数的万用的分子的女伴，用作到例如一种低亲密关系，高能力钙绑定蛋白质，为治疗学的癌症疫苗的一个自然助手，肿瘤拒绝抗原，一个有免疫力的管理者到病理学的房间死亡。它的多功能、结构的特征也做它开发基于抗体的治疗学的一个有趣的目标。然而，因为它在不同种类之中的高顺序的类似，它到老鼠的低 immunogenicity 是一个障碍获得珍贵 monoclonal 抗体(MAbs ) 。这是为任何低产生免疫性的蛋白质的一个普通问题，其序列分享在老鼠和另外的种类之间的靠近的身份。这里， priming 的新策略被猪采用内长的全身的 gp96 然后由 E 增加。coli 系统 heterologously 表示了 gp96 N 终端碎片(N-355 ) 产生 MAbs。对猪 / 人的 12 不同高度特定的 MAbs 内长的 gp96 成功地被获得。这些 MAbs 的有约束力的活动被连接酶的 immunosorbent 证实试金(ELISA ) ，西方的污点(WB ) ， immunofluorescence 和流动 cytometry 分析。这为进一步的研究和潜在的治疗学提供一些重要试剂。采用的方法能被用于在老鼠和猪 / 人之间的任何 sequence-highly-conserved 蛋白质的 MAb 生产(或任何另外的种类) 。

英文摘要：

Gp96, a member of HSP90 family, is a versatile molecular chaperone with various newly-discovered functions, for example to serve as a low affinity, high capacity calcium binding protein, a natural adjuvant for therapeutic cancer vaccines, a tumor rejection antigen, an immune regulator to pathological cell death. Its multi-functional and structural characteristics make it also an interesting target to develop antibody-based therapeutics. However, its low immunogenicity to mice, because of its high-sequence similarity among different species, is an obstacle to obtain valuable monoclonal antibodies （MAbs）. This is a common problem for any low immunogenic proteins, whose sequences share close identity between mice and other species. Here, a new strategy of priming was employed by swine endogenous full-length gp96 and then boosting by E. coli-system heterologously expressed gp96 N-terminal fragment （N-355） to generate MAbs. Twelve different highly-specific MAbs against swine/human endogenous gp96 were successfully obtained. The binding activities of these MAbs were confirmed by enzyme-linked immunosorbent assay （ELISA）, Western blot （WB）, immunofluorescence and flow cytometry analysis. This provides some important reagents for further research and potential therapeutics. The methods employed can be used for MAb production of any sequence-highly-conserved proteins between mice and swine/human （or any other species）.