Preparation and characterization of a neutralizing monoclonal antibody directed to VP4 of rotavirus strain K8 which has unique VP4 neutralization epitopes
Preparation and characterization of a neutralizing monoclonal antibody directed to VP4 of rotavirus strain K8 which has unique VP4 neutralization epitopes
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具有独特VP4中和表位的针对轮状病毒K8株VP4的中和单克隆抗体的制备和表征
DOI：
10.1007/bf01316751
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发表时间：
2005
影响因子：
2.7
通讯作者：
S. Urasawa
中科院分区：
文献类型：
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作者：
Nobumichi Kobayashi;Koki Taniguchi;T. Urasawa;S. Urasawa
为了筛选具有独特中和表位的抗轮状病毒K8株VP 4的中和性单克隆抗体（N-MAbs），我们用K8（血清1型）和P（血清3型）两种不同的轮状病毒株体外混合感染，制备了3株含有K8株VP 4和P株VP 7的重组病毒，4株含有K8株VP 4和P株VP 7的重组病毒。利用这些抑制剂筛选杂交瘤细胞，获得了一株针对K8株特异性VP 4的单克隆抗体（K8- 2C 12）。单克隆抗体K8- 2C 12在针对不同血清型的许多毒株进行检测时仅中和K8，而在酶联免疫吸附试验中，该单克隆抗体也与猴轮状病毒SA 11（血清型3）、牛轮状病毒NCDV（血清型6）和人轮状病毒（HRV）毒株69 M（血清型8）反应。用K8- 2C 12抗体筛选K8抗中和突变株，并测定突变株的VP 4氨基酸序列。在三个突变克隆中检测到394位的单个氨基酸取代，该位置包括在其他轮状病毒中鉴定的主要交叉反应性中和区中。
For selecting the neutralizing monoclonal antibodies (N-MAbs) directed to VP4 of rotavirus strain K8, which has unique VP4 neutralization epitopes, we prepared several reassortant viruses by mixed infection of two different strains K8 (serotype 1) and P (serotype 3) in vitro: three reassortant clones having VP4 of K8 and VP7 of P and four clones having VP4 of P and VP7 of K8. By using these reassortants in screening hybridomas, a N-MAb (K8-2C12) directed to strain K8-specific VP4 was obtained. The MAb K8-2C12 neutralized only K8 when tested against numerous strains of different serotypes, while in enzyme-linked immunosorbent assay this MAb reacted also with simian rotavirus SA11 (serotype 3), bovine rotavirus NCDV (serotype 6), and human rotavirus (HRV) strain 69M (serotype 8). Neutralization-resistant mutants of K8 were selected by the K8-2C12 antibody and VP4 amino acid sequences of the mutants were determined. Single amino acid substitution was detected in the three mutant clones at position 394, which is included in the major cross-reactive neutralization region identified in other rotaviruses.
通过单克隆抗体酶联免疫吸附测定法调查日本不同地区的人类轮状病毒血清型。
DOI：
10.1093/infdis/160.1.44
发表时间：
1989
期刊：
The Journal of infectious diseases
影响因子：
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作者：
S. Urasawa;T. Urasawa;K. Taniguchi;F. Wakasugi;N. Kobayashi;Shunzo Chiba;N. Sakurada;M. Morita;O. Morita;Masayoshi Tokieda;H. Kawamoto;Y. Minekawa;Mitsuaki Ohseto
通讯作者：
Mitsuaki Ohseto
Shozo Urasawa 等人：“通过单克隆抗体 ELISA 对日本不同地区的人类轮状病毒血清型进行调查。”
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