Shan Jin-lu, Dai Nan, Zhang Qin-hong, Li Zeng-peng, Cao Xiao-jing, Wang Dong
Cancer Center, Research Institute of Surgery, Daping Hospital, Third Military Medical University, Chongqing, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010 Mar;26(3):258-60.
To identify the antigenic epitopes of human apurinic/apyrimidinic endonuclease monoclonal antibodies (hAPE1 mAb) and to establish a one-step ELISA for quantitative measurement of hAPE1.
APE1-15 polypeptide array was used to identify the antigenic epitopes of hAPE1 mAb. Molsoft. ICM-Pro software was used to display the three dimensions of antigenic epitopes of hAPE1 mAbs. hAPE1 mAb was labeled with HRP by modified method of sodium periodate oxidization. A one-step ELISA was established by using two hAPE1 mAbs as a capture antibody and a detection antibody, respectively.
The antigenic epitopes of 2-G1 mAb were localized within amino acids (aa) 76-90 and 109-123 in APE1 redox domain, belonging to the conformational epitope category. The antigenic epitope of 4-F6 mAb was within aa 109-147 in DNA repair endonuclease domain. In the range of 8.0 microg/L to 200 microg/L, the hAPE1 antigen showed a good linearity in the standard curve. The minimum detection threshold of this assay was 2.0 microg/L. The average of intra-assay precision and inter-assay precision were 8.67% and 12.45%, respectively. The average recovery rate was 105.47%.
hAPE1 2-G1 mAb and 4-F6 mAb have different antigenic epitopes, and the one-step ELISA is established successfully to detect the serum hAPE1 level easily, quickly and accurately.
鉴定人脱嘌呤/脱嘧啶内切核酸酶单克隆抗体（hAPE1 mAb）的抗原表位，并建立一步酶联免疫吸附测定法（ELISA）定量检测hAPE1。
采用APE1-15多肽芯片鉴定hAPE1 mAb的抗原表位。利用Molsoft. ICM-Pro软件展示hAPE1 mAb抗原表位的三维结构。采用改良的高碘酸钠氧化法将hAPE1 mAb用辣根过氧化物酶（HRP）标记。分别以两种hAPE1 mAb作为捕获抗体和检测抗体，建立一步ELISA。
2-G1 mAb的抗原表位位于APE1氧化还原结构域的氨基酸（aa）76 - 90和109 - 123内，属于构象表位类型。4-F6 mAb的抗原表位位于DNA修复内切核酸酶结构域的aa 109 - 147内。在8.0 μg/L至200 μg/L范围内，hAPE1抗原在标准曲线中呈现良好的线性关系。该检测方法的最低检测限为2.0 μg/L。批内精密度和批间精密度的平均值分别为8.67%和12.45%。平均回收率为105.47%。
hAPE1 2-G1 mAb和4-F6 mAb具有不同的抗原表位，成功建立了一步ELISA，可简便、快速、准确地检测血清hAPE1水平。
