Establishment of a monoclonal antibody against a peptide of the novel zinc finger protein ZNF32 proved to be specific and sensitive for immunological measurements.
Establishment of a monoclonal antibody against a peptide of the novel zinc finger protein ZNF32 proved to be specific and sensitive for immunological measurements.
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针对新型锌指蛋白 ZNF32 肽的单克隆抗体的建立被证明对于免疫学测量具有特异性和敏感性
DOI：
10.12659/msm.882725
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发表时间：
2012-05
期刊：
影响因子：
--
通讯作者：
Wei Y
中科院分区：
文献类型：
--
作者：
Wei Y;Zhang J;Li K;Huang L;Li J;Wang X;Lin P;Wei Y
ZNF32被预测为锌指蛋白，参与细胞分化和肿瘤发生，但其确切功能尚不清楚。特异性单克隆抗体（mab）已广泛应用于研究和临床诊断与治疗。因此，我们建立了抗znf32单抗来表征该蛋白的功能。材料/方法选用ZNF32特异性小肽肽肽49 - 63，合成keyhole帽贝血青素（KLH）肽49 - 63作为抗原免疫小鼠。利用杂交瘤技术制备抗peptide49-63的单抗，通过限制稀释筛选杂交瘤细胞。采用双琼脂扩散法鉴定单克隆抗体- pznf32 - 8d9的同工型。通过酶联免疫吸附试验（ELISA）、免疫细胞化学、免疫组织化学和Western blotting鉴定单抗的敏感性和特异性以及ZNF32在各种细胞和组织中的表达水平。结果建立了一株稳定的抗pznf32 - 8d9杂交瘤，分泌抗肽49 - 63单抗，酶联免疫吸附测定上清抗体阳性克隆率为92%。mAb-pZNF32-8D9是一种免疫球蛋白-1，可通过免疫细胞化学、免疫组织化学和Western blotting检测ZNF32蛋白，具有高度的敏感性和特异性。我们还发现ZNF32在Jurkat和肺鳞癌细胞中高水平表达，但在鳞状表皮细胞中不表达。结论mAb-pZNF32-8D9可用于ZNF32的鉴定和表达。它也可能为znf32相关疾病的诊断或治疗提供新的工具。
Summary Background ZNF32 has been predicted to be a zinc finger protein and is involved in cell differentiation and tumor development, but its precise function is unknown. Specific monoclonal antibodies (mAbs) have been widely used in research and clinical diagnosis and treatments. Therefore, we established an anti-ZNF32 mAb to characterize this protein’s function. Material/Methods Peptide49–63, a specific small peptide of ZNF32, was chosen and the synthetic keyhole limpet hemocyanin (KLH)-peptide49–63 was used as an antigen to immunize mice. A mAb against peptide49–63 was generated by hybridoma technology, and hybridoma cells were screened by limiting dilution. The isoform of mAb-pZNF32-8D9 was identified by double agar diffusion. The sensitivity and specificity of the mAb and expressed levels of ZNF32 in various cells and tissues were identified by enzyme-linked immunosorbent assay (ELISA), immunocytochemistry, immunohistochemistry, and Western blotting. Results A stable anti-pZNF32-8D9 hybridoma secreting the anti-peptide49–63 mAb was established and the clone positive to the peptide49–63 in supernatant was 92% in ELISA. The mAb-pZNF32-8D9 is an immunoglobulin-1 that can be used for detecting the ZNF32 protein by immunocytochemistry, immunohistochemistry, and Western blotting and is highly sensitive and specific. We also found ZNF32 expressed at high levels in Jurkat and pulmonary squamous carcinoma cells, but it was not expressed in squamous epidermis cells. Conclusions mAb-pZNF32-8D9 can be used for the identification and expression of ZNF32. It might also provide a new tool for diagnostics or therapy for ZNF32-related diseases.
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影响因子：
3.7
作者：
Middelbos, Ingmar S.;Vester, Brittany M.;Karr-Lilienthal, Lisa K.;Schook, Lawrence B.;Swanson, Kelly S.
通讯作者：
Swanson, Kelly S.
影响因子：
45.3
作者：
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通讯作者：
Ychou, Marc
影响因子：
3.7
作者：
Liu Q;Sung AH;Chen Z;Liu J;Huang X;Deng Y
通讯作者：
Deng Y
影响因子：
5.4
作者：
Crosnier C;Staudt N;Wright GJ
通讯作者：
Wright GJ
影响因子：
2.7
作者：
Kim, Mihee;Yun, Chul-Ho;Ahn, Taeho
通讯作者：
Ahn, Taeho