分泌抗新疆出血热病毒单克隆抗体杂交瘤细胞株的建立

本文报告了用新疆出血热病毒免疫BALB/c鼠脾脏与骨髓瘤细胞SP2/0融合后建立了6株分泌抗XHF病毒的单克隆抗体杂交瘤细胞。经传代一年余,分泌抗体的稳定性良好,细胞培养液中抗体滴度可达10~(-3);胶水中滴度达10~(-5)～10~(-6)以上。这6株McAb已成功地用于ELISA、IFA和RPHA。在初步研究中观察到分泌IgM免疫球蛋白的14B_7株McAb致敏醛化血球测定病毒抗原性的灵敏度最高;而8E_(12)和13C_6用于间接免疫荧光时其荧光滴度和亮度最好。表明了同一株McAb在不同的血清学试验中所表现出的亲和性及结合力不完全一致。

ESTABLISHMENT OF HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST XINJIANG HEMORRHAGIC FEVER VIRUS

This paper reports the establishment and application of six hybridoma cell lines secreting McAb against XHF virus. The fusion of SP2/0 Ag 14 myeloma cells and splenocytes of BALB/c miceimmunized with BA84002 strain of XHF virus isolated from ticks Hyalomma asiaticum was made according to the routine procedure. Culture supernatants of hybridoma cell were screened by indirect ELISA and indirect immunofluorescent test.These hybridoma cell lines were referred to as 8E12, 10F6, 13C6, 13B7, 17C1 and 43E5. The antibodys-ecreting characters of these hy-bridomas were stadle over one year. The McAb titers of the cultures by ELISA were 10-3 while that of the ascites fluid were higher than 10-5-10-6. The McAbs have been successfully applied to ELISA, IFA and RPHA tests.The McAb of 14B7 was of mouse IgM, in RPHA test determining XHF virus antigens showed high sensitivity whereas the IgG1 of 8E12 and 13C6 expressed high titers in IFA tests indicating that the affinity ofthe same McAb was not parallel in different serological assays.