Barteling S J, Woortmeyer R
Arch Virol. 1984;80(2-3):103-17. doi: 10.1007/BF01310652.
The inactivation of foot-and-mouth disease virus by formaldehyde was studied under different conditions, both as free virus and (as in routine vaccine production) after adsorption of the virus to aluminium hydroxide gel (alhydrogel). In the latter case infectivity was monitored after elution of the virus from the gel by isopycnic ultracentrifugation of the virus-alhydrogel mixture in CsCl. By this method good virus recoveries were obtained. Adsorption of the virus to alhydrogel (without formaldehyde) did not reduce infectivity significantly. Both adsorbed and non-absorbed virus lost infectivity at a rate of about one log10 per day (at pH 8.5, 25 degrees C--no formaldehyde). Kinetics of formaldehyde inactivation of adsorbed and non-adsorbed virus were also identical, with a fast reduction in the initial phase (in case of O1 and A10-virus approximately one log10/hour). After this initial phase inactivation became linear and rather slow (for O1 and A10-virus 0.2 log10/hour). No "tailing-off" was observed. Under standard conditions (0.04 per cent formaldehyde, pH 8.5, 25 degrees C) CD-virus was inactivated approximately 1.5 times faster than O1 and A10-virus. At 4 degrees C the inactivation of the three strains continued at about one log10/day. Increased lactalbumin hydrolysate concentrations reduced the inactivation rate, especially at the formaldehyde concentration of 0.02 per cent, which was originally applied. Quaternary amines like Tris strongly inhibited formaldehyde activity. These findings might explain some data of others who observed "tailing off". Analysis of formaldehyde inactivated antigen by SDS-PAGE and electrofocusing showed that extensive cross-linking occurs especially of VP1, probably with other virus proteins but also with non-virus proteins from the medium. VP2 and VP3 are less affected. Cross-linking was enhanced when the virus had been adsorbed to alhydrogel during inactivation. Progressive cross-linking was observed during storage of the vaccine at 4 degrees C, which also indicated that inactivation continued at this temperature. These data show that formaldehyde inactivated adsorbate vaccines can be safe.
研究了在不同条件下甲醛对口蹄疫病毒的灭活作用，包括游离病毒以及（如在常规疫苗生产中）病毒吸附到氢氧化铝凝胶（铝胶）后的情况。在后一种情况下，通过在氯化铯中对病毒 - 铝胶混合物进行等密度超速离心从凝胶中洗脱病毒后监测其感染性。通过这种方法可获得良好的病毒回收率。病毒吸附到铝胶上（无甲醛）不会显著降低感染性。吸附和未吸附的病毒均以约每天一个对数 10 的速率丧失感染性（在 pH 8.5、25℃ - 无甲醛条件下）。吸附和未吸附病毒的甲醛灭活动力学也相同，在初始阶段快速降低（对于 O1 和 A10 病毒，约每小时一个对数 10）。在此初始阶段之后，灭活变得线性且相当缓慢（对于 O1 和 A10 病毒为每小时 0.2 个对数 10）。未观察到“拖尾”现象。在标准条件（0.04%甲醛，pH 8.5，25℃）下，CD 病毒的灭活速度比 O1 和 A10 病毒快约 1.5 倍。在 4℃时，三种毒株的灭活以约每天一个对数 10 的速度继续。增加水解乳白蛋白浓度可降低灭活速率，尤其是在最初使用的 0.02%甲醛浓度下。像 Tris 这样的季胺强烈抑制甲醛活性。这些发现可能解释了其他一些观察到“拖尾”现象的数据。通过 SDS - PAGE 和等电聚焦对甲醛灭活抗原的分析表明，尤其是 VP1 发生了广泛的交联，可能与其他病毒蛋白以及培养基中的非病毒蛋白交联。VP2 和 VP3 受影响较小。当病毒在灭活过程中吸附到铝胶上时，交联增强。在疫苗于 4℃储存期间观察到交联逐渐增加，这也表明在此温度下灭活仍在继续。这些数据表明甲醛灭活的吸附疫苗可能是安全的。
