CAJ | 학술논문

目的：：筛选抗脂多糖（LPS）纳米单域抗体，并制备抗LPS纳米抗体五聚体。方法：以LPS为抗原，从驼源天然单域重链抗体库中筛选抗LPS纳米抗体，利用分子克隆技术将抗LPS单域抗体基因组装入志贺杆菌样毒素B亚基蛋白结构域（VTB）的五聚体特异性表达载体中进行可溶性表达，并用ELISA法鉴定所获抗体的抗原结合活性和特异性。结果：获得抗LPS纳米单域抗体及LPS纳米抗体五聚体；经鉴定，LPS纳米抗体五聚体的抗原结合活性优于抗LPS单域抗体。结论：利用驼源天然单域重链抗体库制备了抗LPS纳米单域抗体及抗LPS纳米抗体五聚体，为脓毒血症的分子诊断、预后判断及寻找生物治疗新靶点奠定了基础。
목적：：사선항지다당（LPS）납미단역항체，병제비항LPS납미항체오취체。방법：이LPS위항원，종타원천연단역중련항체고중사선항LPS납미항체，이용분자극륭기술장항LPS단역항체기인조장입지하간균양독소B아기단백결구역（VTB）적오취체특이성표체재체중진행가용성표체，병용ELISA법감정소획항체적항원결합활성화특이성。결과：획득항LPS납미단역항체급LPS납미항체오취체；경감정，LPS납미항체오취체적항원결합활성우우항LPS단역항체。결론：이용타원천연단역중련항체고제비료항LPS납미단역항체급항LPS납미항체오취체，위농독혈증적분자진단、예후판단급심조생물치료신파점전정료기출。
Objective: To obtain anti-LPS VHH and prepare anti-LPS nanobody pentamer. Methods: Using phage display screening technology, we selected anti-LPS single domain antibody(sdAb, or nanobody) from a phage display library of llama single domain antibody, and then cloned the anti-LPS single-domain antibody gene into specific expression vector pVTB1 with molecular cloning technology, and expressed anti-LPS pentamer effi-ciently in E.coli. The affinity of different type of anti-LPS nanobodies were detected by ELISA. Results: We ob-tained anti-LPS single domain antibody and anti-LPS antibody pentamer, and proved anti-LPS nanobody pentamer is extremely enhance the specificity and affinity strength of anti-LPS antibody. Conclusion: The selected anti-LPS antibody pentamer is a leading candidate for therapies against LPS-mediated sepsis.