Quantitation of group-specific a antigen in hepatitis B vaccines by anti-HBs/a monoclonal antibody.
Quantitation of group-specific a antigen in hepatitis B vaccines by anti-HBs/a monoclonal antibody.
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通过抗 HBs/a 单克隆抗体对乙型肝炎疫苗中的组特异性 a 抗原进行定量。
DOI：
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发表时间：
1997
期刊：
影响因子：
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通讯作者：
Y. Moritsugu
中科院分区：
文献类型：
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作者：
H. Yamamoto;T. Satoh;T. Kiyohara;A. Totsuka;Y. Moritsugu
用氢氧化铝[alum，Al（OH）3]佐剂的adr、ayw或adw亚型B（HB）肝炎疫苗免疫BalB/c小鼠。免疫动物脾细胞与SP2/O细胞融合。筛选出8株能产生抗乙肝表面抗原（HBsAg）特异性抗体的杂交瘤细胞株。4个克隆的单克隆抗体（mAb）对群特异性抗原/a具有特异性，而4个克隆的另一个对亚型抗原/d、y、r或w具有特异性。抗-HBs/a单克隆抗体分为三个非竞争性组。用抗-HBs豚鼠多克隆抗体（pAb）固相、检测用HBsAg、抗-HBs/a鼠单克隆抗体（mAb）和辣根过氧化物酶（HRP）标记的抗-鼠IgG，用夹心酶联免疫吸附试验（ELISA）定量检测HBsAg的组特异性决定簇a（HBsAg/a）。用未吸附的HBsAg建立HBsAg/a标准曲线。HBsAg的检测下限为0.5 ~ 1 ng/ml。研究了明矾增溶法定量吸附乙肝疫苗中HBsAg/a的方法。疫苗经预稀释后，回收率可达60%以上。同一厂家生产的乙肝疫苗中HBsAg/a的回收率相似，吸附乙肝疫苗中HBsAg/a的含量可通过测定吸附乙肝疫苗的回收率来估计。
Balb/c mice were immunized with aluminium hydroxide [alum, Al (OH)3]-adjuvanted hepatitis B (HB) vaccines of subtypes adr, ayw or adw. Spleen cells from the immune animals were fused with SP2/O cells. Eight hybridoma clones producing antibodies specific for HB surface antigen (HBsAg) were selected. Monoclonal antibodies (mAbs) of four clones were specific for group-specific antigen/a, and the other of four clones were specific for subtype antigen/d, y, r, or w. The anti-HBs/a mAbs were classified into three non-competitive groups. Quantitation of group-specific determinant a of HBsAg (HBsAg/a) was performed by sandwich enzyme-linked immunosorbent assay (ELISA), in which a solid phase of anti-HBs guinea-pig polyclonal antibodies (pAb), the HBsAg for testing, anti-HBs/a mouse mAb and horseradish peroxidase (HRP)-conjugated anti-mouse IgG were used. The unadsorbed HBsAg was used to establish the standard curve of HBsAg/a. The lower detection limits were 0.5 to 1 ng/ml of HBsAg. Methods of solubilization of alum were investigated to quantify HBsAg/a in adsorbed HB vaccines. The recovery rate was more than 60% if vaccines were prediluted. The recovery of HBsAg/a in HB vaccines produced by the same manufacturer showed the similar recovery rate, and the contents of HBsAg/a in adsorbed HB vaccines could be estimated by the recovery rate determined for adsorbed HB vaccines.