中文摘要：

为研制抗三肽囊素（Bursin，KHG．NH2）单克隆抗体（MAb），本研究采用EDC法、SMCC法将Bursin分子赖氨酰（K）上的氨基与载体蛋白（BSA、OVA）定向偶联，分别制备Bursin人工抗原BSA．KHG-NH2、OVA-KHG-NH2和BSA-S—KHG—NH2。以BSA—KHG．NH2免疫BALB／c小鼠，应用杂交瘤技术，筛选得到二株能够稳定分泌抗BursinMAb的杂交瘤细胞株，命名为4A11和782。这二株MAbs均能够与BSA．KHG．NH，OVA—KHG—NH2和BSA-S-KHG—NH2反应，不能与BSA和OVA反应；MAbs与OvA．KHG．NH2的反应均能够被KHG-NH2、GKHG-NH2、HHHHHHKHG-NH2肽阻断，但不能被KHGK四肽阻断。结果表明，在Bursin分子甘氨酰胺（G）上的氨基未与其他氨基酸连接时，本实验制备的二株MAbs均能够与Bursin分子特异性结合。抗BursinMAbs的制备，为进一步以MAb为靶分子筛选Bursin模拟表位、Bursin的生物活性关键基团等研究奠定了基础。

英文摘要：

To prepare the monoclonal antibody （MAb） against the bursin and identified the binding sit of bursin, the bursin were conjugated to bovine serum albumin （BSA） and chicken ovalbumin （OVA） through the amino group on -Lys of Bursin by EDC and SMCC methods to prepare the artificial antigen of bursin （BSA-KHG-NH2, OVA-KHG-NI-I= and BSA-S-KHG-NH2）, and the BSA-KHG-NH2 was used to immunize BALB/c mice. Hybridomas were prepared from the mice, which were demonstrated to produce MAbs against Bursin by indirect ELISA. The two MAbs bond specifically to the BSA-KHG-NH2, OVA-KHG-NH2 and BSA-S-KHG-NH2 conjugates. In addition, the binding of MAb 4A11 and 7B2 to OVA-KHG-NH2 were competitively inhibited by free KHG-NH2, GKHG-NH2 and HHHHHHKHG-NH2 peptides, respectively, but unable to be inhibited by free KHGK peptide. These results strongly suggested that these two MAbs bound to the structural epitope of bursin with the amino group on -Gly.