Matsuda M, Kamei M, Sugimoto N, Ma Y, Hashizume S
Department of Tuberculosis Research, Osaka University, Japan.
Eur J Epidemiol. 1992 Jan;8(1):1-8. doi: 10.1007/BF02427384.
Five anti-tetanus human monoclonal antibodies (MAbs) produced by hybrid cell lines we established previously were characterized. Their abilities to neutralize tetanus toxin in vitro and to protect mice against challenge with toxin were studied by observing the changes in the progress of symptoms in mice. Immunostaining showed that MAbs MAb-G4 and G2 recognized the N-terminal domain, [A] and the C-terminal domain, [C] of the tetanus toxin molecule, respectively, while MAbs MAb-G1, G3 and G6 recognized its middle domain, [B]. Enzyme-linked immunosorbent assay showed that the binding affinity of MAb-G3 was 2.9 x 10(10) M-1 and those of the other MAbs were as high as approximately 10(11) M-1. In in vitro neutralization experiments, at sufficient doses all the MAbs as single reagents protected mice completely against the effect of tetanus toxin. However, at lower doses than those sufficient to rescue mice, the kinetic patterns of progress of symptoms with the individual MAbs differed with each other and, except for MAb-G4, were different from that of anti-tetanus human polyclonal antibody. They suppressed the development and/or slowed the rate of progress of symptoms for over 96 h and delayed death of the mice. We propose that the comparison of the minimum survival dose with that of human polyclonal antibody of known international units is a reliable method for estimating the actual protective activity of a MAb. Intravenous (IV) injection of doses of individual MAbs or their mixtures at over 0.03 IU per mouse protected mice from subsequent challenge with 20 MLD of tetanus toxin.(ABSTRACT TRUNCATED AT 250 WORDS)
对我们之前建立的杂交细胞系产生的5种抗破伤风人单克隆抗体（MAb）进行了特性分析。通过观察小鼠症状进展的变化，研究了它们在体外中和破伤风毒素以及保护小鼠免受毒素攻击的能力。免疫染色显示，单克隆抗体MAb-G4和G2分别识别破伤风毒素分子的N端结构域[A]和C端结构域[C]，而单克隆抗体MAb-G1、G3和G6识别其中间结构域[B]。酶联免疫吸附测定表明，单克隆抗体G3的结合亲和力为2.9×10¹⁰ M⁻¹，其他单克隆抗体的结合亲和力高达约10¹¹ M⁻¹。在体外中和实验中，所有单克隆抗体作为单一试剂，在足够剂量下能完全保护小鼠免受破伤风毒素的影响。然而，在低于足以挽救小鼠的剂量下，各单克隆抗体使症状进展的动力学模式彼此不同，除了MAb-G4外，均与抗破伤风人多克隆抗体的模式不同。它们抑制症状的发展和/或减缓症状进展速度超过96小时，并延迟小鼠死亡。我们提出，将最低存活剂量与已知国际单位的人多克隆抗体的剂量进行比较，是估计单克隆抗体实际保护活性的可靠方法。每只小鼠静脉注射（IV）超过0.03 IU的各单克隆抗体或其混合物，可保护小鼠免受随后20 MLD破伤风毒素的攻击。（摘要截短于250字）
