一种人凝血因子的制作方法

一种人凝血因子
ⅸ
的干热灭活装置及工艺
技术领域
1.本发明涉及干热灭活技术领域，具体涉及一种人凝血因子
ⅸ
的干热灭活装置及工艺。


背景技术：

2.人凝血因子
ⅸ
是一种维生素k依赖的单链糖蛋白，在凝血过程中起重要作用，缺乏或缺失人凝血因子
ⅸ
会导致乙型血友病。20世纪90年代起，不含fⅱ、fⅶ、f
ⅹ
等杂蛋白的高纯人凝血因子
ⅸ
制品开始研制，人凝血因子
ⅸ
比活性可达50-200iu/mg，是目前治疗乙型血友病的最佳选择。
3.为了保障血液制品的安全性，现阶段基本采用s/d法（有机溶剂/去污剂混合物法）结合干热灭活法的两次病毒灭活步骤对人凝血因子
ⅸ
制品进行处理。其中，现有的凝血因子类干热灭活工艺大多采用冻干品80℃、72h干热灭活或100℃、30min干热灭活，但这两种干热灭活工艺均存在一定缺陷，很难在灭活效果达标的同时获得理化性质、生物学活性及稳定性均合格的制品。对于80℃、72h的干热灭活工艺，其存在灭活时间长、制品效价损失高的缺陷。对于100℃、30min的干热灭活工艺，由于升温和冷却过程没有进行有效控制，整个干热灭活工序时间难以控制，随着制品升温和冷却时间的延长，制品存在效价的损失；同时在实际批量进行干热灭活操作时，难以控制将灭活温度始终保持在100℃。


技术实现要素：

4.针对现有干热灭活工艺的干热灭活时间长、升温及冷却阶段效价损失、灭活温度不易控制的技术问题，本发明提供一种人凝血因子
ⅸ
的干热灭活装置及工艺，解决了现有的干热灭活工艺工序时间长，升温段及冷却段难以有效控制，从而造成人凝血因子
ⅸ
的效价损失的问题，本发明灭活段温度更容易控制，病毒灭活效果好，干热灭活后效价损失低，具有很大的应用价值。
5.第一方面，本发明提供一种人凝血因子
ⅸ
的干热灭活装置，包括灭活内室，灭活内室分别连通压缩空气管线、排汽管线、注水管线和排泄管线，压缩空气管线设有压缩空气阀，排汽管线设有排汽阀，注水管线设有注水阀，排泄管线设有排泄阀；灭活内室顶部和底部分别设有温度传感器，灭活内室顶部还设有压力传感器；灭活内室顶部通过换热器与蒸汽管线、冷却水管线连通，灭活内室顶部与换热器间的连通管线上设有温度传感器，蒸汽管线包括第一蒸汽子管线和第二蒸汽子管线，第一蒸汽子管线设有大蒸汽阀，第二蒸汽子管线设有小蒸汽阀，冷却水管线包括第一冷却水子管线和第二冷却水子管线，第一冷却水子管线设有大冷却水阀，第二冷却水子管线设有小冷却水阀，此处的“大”、“小”为相对概念，分别依据两个蒸汽阀的最大流通面积和两个冷却水阀的最大流通面积确定；灭活内室底部通过换热器与排水管线、疏水管线和排冷凝水管线连通，排水管线设有排冷却水阀，疏水管线设有疏水阀，排冷凝水管线设有冷凝水旁通阀，灭活内室底部与换热器间的连通管线上设有温度传感器。
6.进一步的，由于温度对于人凝血因子
ⅸ
效价影响巨大，灭活室内底部设置至少两个温度传感器，优选为四个，防止单个传感器温度监测失效的问题；灭活室内顶部设置至少两个压力传感器，防止单个压力传感器故障造成的压力失真。
7.进一步的，小蒸汽阀为比例调节阀，升温阶段温度控制更加精细，温度上升快速均匀。
8.进一步的，灭活室底部与换热器的连通管线上还设有循环泵。
9.进一步的，干热灭活装置还包括控制器，温度传感器、压力传感器、大蒸汽阀、小蒸汽阀、大冷却水阀、小冷却水阀、压缩空气阀、排汽阀、注水阀、排泄阀、排冷却水阀、疏水阀、冷凝水旁通阀分别与控制器电性连接，控制器还与显示器电性连接，显示器用于显示温度传感器和压力传感器的检测数值及阀门的开关状态。
10.进一步的，大蒸汽阀、大冷却水阀、小冷却水阀、压缩空气阀、排汽阀、注水阀、排泄阀、排冷却水阀、疏水阀、冷凝水旁通阀为气动阀。
11.第二方面，本发明提供一种人凝血因子
ⅸ
的干热灭活工艺，采用上述干热灭活装置，步骤如下：s1注水；s2升温：开始升温时大蒸汽阀间歇打开，小蒸汽阀开度调节至100%，升温速度为6.0-7.5℃/s，当灭活内室底部的温度传感器温度到达95.0℃后大蒸汽阀关闭，当灭活内室底部的温度传感器温度到达97.0℃后，小蒸汽阀开度调节至80%，当灭活内室底部的温度传感器温度到达97.5℃时通压缩空气，使灭活内室压力到达160kpa并维持压力在160-165kpa，疏水阀、冷凝水旁通阀开启；s3灭活：当灭活内室底部的温度传感器温度到达98.2℃，小蒸汽阀开度调节至50%，当灭活内室底部的温度传感器温度到达98.6℃，小蒸汽阀开度调节至15%且间歇开启，使灭活温度保持在98.2-99.2℃，间歇通入压缩空气使灭活内室压力保持在160-165kpa，灭活时间为1800s；s4冷却：疏水阀、冷凝水旁通阀关闭，间歇通入压缩空气使灭活内室压力保持在155-160kpa，小冷却水阀打开30s、关闭30s，当灭活内室底部的温度传感器温度到达90.0℃后，排冷却水阀打开，大冷却水阀与小冷却水阀同时打开30s、关闭30s，当灭活内室底部的温度传感器温度低于50.0℃冷却结束；s5排水、排汽。
12.进一步的，s1注水步骤为：打开注水阀，注入去离子水，去离子水的液位控制在150-300mm之间，保证了灭活柜内高温水循环的最低液位，由于高温水循环使用，会接触到模制瓶的外壁，需要保证一定的洁净度，去离子水的电导率小于60us/cm。
13.进一步的，灭活内室底部的温度传感器数量大于一个，s2升温步骤中依据灭活内室底部各个温度传感器的最低温度控制大蒸汽阀开闭及小蒸汽阀开度。
14.进一步的，干热灭活工艺的处理量为2000-6000瓶/批。
15.进一步的，s2升温步骤小于1000s，s4冷却步骤小于600s，防止升温、冷却时间过长造成的效价损失。
16.进一步的，s5排水、排汽步骤为：排汽阀打开，排泄阀打开，灭活内室顶部压力传感
器的压力低于10kpa时排水、排汽结束。
17.进一步的，灭活内室顶部压力传感器数量大于一个，s5排水、排汽步骤中，灭活内室顶部各个压力传感器的压力均低于10kpa时排水、排汽结束。
18.本发明的有益效果在于：本发明通过控制升温及冷却的时间，减少了制品升温及冷却阶段的效价损失，整个干热灭活时间短，升温及冷却阶段采用了双阀控制，升温阶段提升了升温速度，同时保证制品升温的均匀，灭活温度保持在98.2-99.2℃，更加容易控制。
附图说明
19.为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，对于本领域普通技术人员而言，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
20.图1是实施例1人凝血因子
ⅸ
的干热灭活装置的结构示意图。
21.图2是实施例2中2019s02批人凝血因子
ⅸ
的干热灭活趋势图。
22.图3是实施例2中2019s03批人凝血因子
ⅸ
的干热灭活趋势图。
23.图4是实施例2中2019s04批人凝血因子
ⅸ
的干热灭活趋势图。
24.图5是实施例3干热法灭活后人凝血因子
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样品中ppv动力学曲线。
25.图6是实施例3干热法灭活后人凝血因子
ⅸ
样品中emcv动力学曲线。
26.图中，1-小蒸汽阀fc，2-大蒸汽阀fb，3-小冷却水阀fe，4-大冷却水阀fd，5-压力传感器pa，6-压力传感器pb，7-温度传感器th，8-压缩空气阀fa，9-排汽阀fj，10-注水阀ff，11-排泄阀fg，12-温度传感器tl，13-循环泵，14-换热器，15-排冷却水阀fi，16-疏水阀fh，17-冷凝水旁通阀fk，18-温度传感器ta，19-温度传感器tb，20-温度传感器tc，21-温度传感器td，22-温度传感器te。
具体实施方式
27.为了使本技术领域的人员更好地理解本发明中的技术方案，下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都应当属于本发明保护的范围。
28.实施例1一种人凝血因子
ⅸ
的干热灭活装置，包括灭活内室和控制器，灭活内室分别连通压缩空气管线、排汽管线、注水管线和排泄管线，压缩空气管线设有压缩空气阀，排汽管线设有排汽阀，注水管线设有注水阀，排泄管线设有排泄阀；灭活内室顶部设有温度传感器te22、压力传感器pa5和压力传感器pb6，灭活内室底部设有温度传感器ta18、温度传感器tb19、温度传感器tc20和温度传感器td21；灭活内室顶部通过换热器14与蒸汽管线、冷却水管线连通，灭活内室顶部与换热器14间的连通管线上设有温度传感器th7，蒸汽管线包括第一蒸汽子管线和第二蒸汽子管线，第一蒸汽子管线设有大蒸汽阀fb2，第二蒸汽子管线设有小蒸汽阀fc1，冷却水管线包括第一冷却水子管线和第二冷却水子管线，第一冷却水子管线
设有大冷却水阀fd4，第二冷却水子管线设有小冷却水阀fe3，此处的“大”、“小”为相对概念，分别依据两个蒸汽阀的最大流通面积和两个冷却水阀的最大流通面积确定；灭活内室底部通过换热器14与排水管线、疏水管线和排冷凝水管线连通，灭活室底部与换热器14的连通管线上还设有循环泵13，排水管线设有排冷却水阀fi15，疏水管线设有疏水阀fh16，排冷凝水管线设有冷凝水旁通阀fk17，灭活内室底部与换热器14间的连通管线上设有温度传感器tl12；温度传感器ta18，温度传感器tb19，温度传感器tc20，温度传感器td21，温度传感器te22、温度传感器th7、温度传感器tl12、压力传感器pa5、压力传感器pb6、大蒸汽阀fb2、小蒸汽阀fc1、大冷却水阀fd4、小冷却水阀fe3、压缩空气阀fa8、排汽阀fj9、注水阀ff10、排泄阀fg11、排冷却水阀fi15、疏水阀fh16、冷凝水旁通阀fk17分别与控制器电性连接，控制器还与显示器电性连接，显示器用于显示各温度传感器和压力传感器的检测数值及阀门的开关状态；大蒸汽阀fb2、大冷却水阀fd4、小冷却水阀fe3、压缩空气阀fa8、排汽阀fj9、注水阀ff10、排泄阀fg11、排冷却水阀fi15、疏水阀fh16、冷凝水旁通阀fk17为气动阀；小蒸汽阀fc1为比例调节阀（盖米514s-25d-137-51-1）。
29.实施例2采用实施例1的干热灭活装置对人凝血因子
ⅸ
进行干热灭活，步骤如下：s1注水：打开注水阀ff10，注入去离子水，去离子水的液位控制在150-300mm之间，电导率小于60us/cm；s2升温：开始升温时大蒸汽阀fb2间歇打开，小蒸汽阀fc1开度调节至100%，升温速度为6.0-7.5℃/s，此时灭活内室内温度快速升高，当温度传感器ta18、温度传感器tb19、温度传感器tc20和温度传感器td21中的最低温度（以下简称“最低温度”）到达95.0℃后大蒸汽阀fb2关闭，避免温度升高过快，造成人凝血因子
ⅸ
升温不均匀；当最低温度到达97.0℃后，小蒸汽阀fc1开度调节至80%，通过小蒸汽阀fc1开度的调节，保证人凝血因子
ⅸ
升温的均匀；当最低温度到达97.5℃时，压缩空气阀fa8打开，使灭活内室压力到达160kpa，压缩空气阀fa8间歇打开，使灭活内室压力维持在160-165kpa，疏水阀fh16、冷凝水旁通阀fk17开启；升温阶段总时间小于1000s；如图2-4所示，升温段为平滑的曲线，温度上升均匀快速；s3灭活：当最低温度到达98.2℃，小蒸汽阀fc1开度调节至50%，当最低温度到达98.6℃，小蒸汽阀开度调节至15%且间歇开启，使灭活温度保持在98.2-99.2℃，压缩空气阀fa8间歇打开，使灭活内室压力保持在160-165kpa，灭活时间为1800s，此时小蒸汽阀fc1间歇通入少量蒸汽便满足温度变化需求，如图2-4所示，灭活段温度平稳；s4冷却：疏水阀fh16、冷凝水旁通阀fk17关闭，压缩空气阀fa8间歇打开，使灭活内室压力保持在155-160kpa，小冷却水阀fe3打开30s、关闭30s，小冷却水阀fe3打开时温度快速下降，小冷却水阀fe3关闭时温度无明显变化；当最低温度到达90.0℃后，排冷却水阀fi15打开，大冷却水阀fd4与小冷却水阀fe3同时打开30s、关闭30s，在双阀开启的过程中温度快速下降，当最低温度低于50.0℃冷却结束，如图2-4所示冷却段温度阶梯变化，降温速度快；s5排水、排汽：排汽阀fj9打开，排泄阀fg11打开，当压力传感器pa5、压力传感器pb6的压力低于10kpa时排水、排汽结束。
30.上述干热灭活工艺过程中，显示器实时显示各点的温度、压力数值和各阀门的开关状态。
31.根据上述干热灭活工艺进行三批次试验，其中2019s02批（共2574瓶）干热灭活数据如下表1，2019s03批（共2657瓶）干热灭活数据如下表2，2019s04批（共2620瓶）干热灭活数据如下表3，升温阶段每分钟采集一次数据，灭活阶段每30s采集一次数据，冷却阶段每分钟采集一次数据。
32.表1：2019s02批干热灭活数据（℃、kpa）时间tatbtctd平均温度papbtlthte15:10:1823.523.624.124.023.80.00.024.423.623.715:10:2823.523.624.124.023.80.00.024.423.623.715:11:1830.831.031.031.131.04.04.027.930.629.915:12:1837.537.637.637.737.68.08.035.337.436.415:13:1844.444.544.644.744.613.013.042.144.443.115:14:1851.451.551.451.751.520.019.049.151.450.315:15:1858.058.158.258.258.127.026.055.858.056.915:16:1864.564.564.664.764.635.034.062.364.663.415:17:1870.770.770.870.970.845.044.068.670.869.615:18:1876.776.676.877.076.856.055.074.776.976.115:19:1881.981.882.082.282.068.067.080.382.181.215:20:1886.686.586.887.086.779.079.084.986.985.915:21:1891.691.591.892.091.794.094.089.992.091.015:22:1896.296.196.596.696.4110.0110.094.696.795.915:23:1897.697.998.799.098.3161.0161.097.498.197.315:24:1898.198.198.798.798.5162.0163.098.298.598.315:24:4498.398.398.999.098.8164.0164.098.298.898.515:24:4898.998.799.199.199.0164.0164.098.298.798.615:25:1898.798.698.999.098.8163.0164.098.398.998.615:25:4898.698.498.998.998.7164.0164.098.498.698.615:26:1898.998.799.199.199.0164.0164.098.498.798.815:26:4898.898.699.099.198.9163.0164.098.498.898.615:27:1898.798.598.999.098.8163.0164.098.498.698.715:27:4898.898.799.099.198.9164.0164.098.498.998.815:28:1898.998.799.199.199.0164.0164.098.498.798.915:28:4898.798.598.998.998.8164.0164.098.498.698.815:29:1899.098.899.199.299.0164.0164.098.498.998.815:29:4899.098.899.299.199.0164.0164.098.498.799.015:30:1898.998.899.199.199.0164.0164.098.498.798.915:30:4898.798.699.099.098.8164.0164.098.598.698.815:31:1898.798.799.099.098.9164.0164.098.598.798.9
15:31:4898.898.799.099.098.9164.0164.098.598.798.915:32:1898.898.799.099.098.9164.0164.098.598.698.815:32:4898.898.799.099.098.9163.0164.098.598.798.915:33:1898.898.799.099.098.9164.0164.098.598.798.915:33:4898.898.799.099.098.9164.0164.098.598.698.915:34:1898.898.799.099.098.9164.0164.098.598.798.915:34:4898.898.799.099.098.9164.0164.098.598.798.915:35:1899.098.899.199.199.0164.0165.098.598.799.015:35:4899.098.999.299.299.1164.0165.098.598.998.915:36:1898.898.798.999.098.9164.0164.098.598.898.815:36:4898.898.798.998.998.8164.0164.098.598.698.815:37:1898.898.799.099.098.9164.0164.098.598.698.815:37:4898.998.799.099.098.9164.0164.098.598.798.915:38:1899.099.099.299.299.1164.0165.098.598.899.015:38:4898.898.798.998.998.8164.0164.098.598.798.815:39:1898.898.798.999.098.9164.0164.098.598.698.915:39:4898.998.799.099.098.9164.0164.098.598.698.915:40:1898.998.899.099.098.9164.0164.098.598.798.915:40:4898.898.798.998.998.8164.0164.098.598.798.815:41:1898.898.899.099.098.9164.0164.098.598.698.815:41:4898.998.899.099.098.9164.0164.098.598.798.915:42:1899.199.099.299.299.1165.0165.098.598.899.015:42:4898.898.798.998.998.8164.0164.098.598.698.815:43:1898.898.899.099.098.9164.0164.098.598.698.915:43:4898.998.899.199.199.0164.0165.098.598.799.015:44:1898.898.798.999.098.9164.0164.098.598.698.915:44:4898.998.899.099.098.9163.0164.098.598.798.915:45:1899.199.099.199.299.1164.0165.098.598.998.915:45:4898.898.899.099.098.9164.0164.098.598.698.915:46:1898.998.899.099.098.9164.0164.098.598.799.015:46:4898.898.898.999.098.9164.0164.098.598.698.915:47:1898.998.899.099.098.9164.0164.098.598.698.915:47:4899.098.999.199.199.0164.0165.098.598.799.015:48:1898.898.899.099.098.9164.0164.098.598.698.915:48:4898.998.899.099.098.9164.0165.098.598.799.015:49:1898.998.899.099.098.9164.0164.098.598.698.915:49:4898.998.899.099.098.9164.0164.098.598.798.915:50:1898.898.898.999.098.9164.0164.098.598.698.915:50:4898.998.899.099.098.9164.0164.098.698.798.9
15:51:1898.898.898.999.098.9164.0164.098.598.698.915:51:4898.998.899.099.098.9164.0164.098.698.798.915:52:1898.998.898.999.098.9164.0164.098.598.698.915:52:4898.998.899.099.098.9164.0164.098.698.798.915:53:1898.898.898.999.098.9164.0164.098.598.698.915:53:4898.998.899.099.098.9164.0165.098.698.798.915:54:1898.998.898.998.998.9164.0164.098.598.698.915:54:4498.998.899.099.098.9164.0164.098.598.698.915:55:1891.691.392.092.291.8162.0162.095.191.691.315:56:1886.686.486.586.686.5158.0158.090.385.687.215:57:1879.278.779.179.379.1159.0161.083.479.178.815:58:1871.971.971.972.172.0156.0157.077.172.373.015:59:1866.866.566.966.966.8159.0161.071.366.567.516:00:1862.462.362.462.562.4159.0159.066.261.963.016:01:1858.458.458.458.458.4157.0158.061.657.959.316:02:1854.954.954.954.954.9159.0160.057.554.455.516:03:1851.851.651.851.751.7160.0161.053.951.252.216:04:1549.049.049.049.149.0156.0156.050.848.549.916:04:1848.948.948.848.948.9153.0153.050.648.449.416:05:1847.247.246.046.346.795.095.048.848.250.316:06:1845.346.144.144.645.054.055.048.948.349.516:07:1845.846.444.244.745.328.028.048.848.449.016:08:1845.946.644.745.245.612.012.049.048.748.916:08:3046.246.745.145.445.910.010.049.048.849.0由表1可以看出2019s02批升温开始时间为15:10:28，灭菌开始时间为15:24:44，冷却开始时间为15:54:44，冷却结束时间为16:04:15，排水、排汽结束时间为16:08:30。
33.表2：2019s03批干热灭活数据（℃、kpa）时间tatbtctd平均温度papbtlthte14:52:2623.723.724.124.123.90.00.024.623.923.614:52:3623.723.724.124.123.90.00.024.623.923.614:53:2630.330.430.430.530.44.04.028.330.129.414:54:2637.837.937.837.937.98.08.035.837.636.114:55:2644.244.344.244.444.313.013.042.444.142.614:56:2650.250.250.350.450.318.018.048.550.149.214:57:2655.955.956.056.156.024.024.054.255.954.514:58:2661.361.361.461.561.430.030.059.761.459.914:59:2666.566.566.766.766.638.037.065.066.665.715:00:2671.671.771.871.971.846.046.070.271.970.615:01:2676.776.777.077.076.956.055.075.377.075.9
15:02:2681.581.581.881.881.766.066.080.381.881.115:03:2686.386.386.686.786.578.078.085.086.785.915:04:2690.990.991.291.291.192.092.089.891.390.615:05:2695.094.995.295.295.1105.0105.093.495.294.615:06:2697.096.997.397.497.2113.0113.096.297.396.815:07:2698.198.298.798.898.5162.0163.097.698.598.215:08:2298.598.498.898.898.6164.0164.098.298.598.615:08:5698.898.799.199.098.9164.0164.098.398.798.815:09:2698.898.699.099.198.9164.0164.098.498.998.715:09:5698.698.498.998.998.7164.0164.098.498.798.715:10:2698.898.699.099.098.9164.0164.098.498.798.815:10:5698.798.599.099.098.8164.0164.098.498.898.715:11:2698.898.699.099.098.9164.0165.098.498.798.815:11:5698.798.598.998.998.8164.0164.098.598.798.715:12:2698.998.799.199.199.0165.0165.098.498.798.915:12:5698.798.699.099.098.8164.0165.098.598.698.815:13:2698.798.698.998.998.8164.0164.098.598.698.715:13:5698.998.899.199.199.0165.0165.098.498.898.915:14:2698.898.699.099.098.9164.0165.098.598.798.815:14:5698.898.699.099.098.9164.0165.098.598.698.815:15:2698.798.699.099.098.8164.0165.098.598.698.815:15:5698.898.699.099.098.9164.0165.098.598.698.815:16:2698.798.698.999.098.8164.0165.098.598.698.815:16:5698.898.799.099.098.9165.0165.098.598.698.815:17:2698.898.799.099.098.9164.0165.098.598.698.815:17:5698.898.698.999.098.8164.0165.098.598.698.815:18:2698.898.799.099.098.9164.0165.098.598.698.815:18:5698.898.798.999.098.9164.0165.098.598.698.815:19:2698.898.799.099.098.9165.0165.098.598.698.815:19:5698.998.799.099.098.9165.0165.098.598.798.915:20:2699.199.099.299.299.1165.0165.098.598.899.015:20:5698.898.798.998.998.8164.0165.098.598.698.815:21:2698.898.799.099.098.9164.0165.098.598.698.815:21:5698.898.799.099.098.9165.0165.098.598.698.915:22:2698.998.899.099.098.9165.0165.098.598.799.015:22:5699.199.099.299.299.1165.0166.098.598.899.115:23:2698.898.798.999.098.9164.0165.098.598.698.815:23:5698.998.799.099.098.9165.0165.098.598.698.915:24:2698.998.899.099.098.9165.0165.098.598.799.0
15:24:5698.898.799.099.098.9164.0165.098.598.698.915:25:2698.998.899.099.098.9165.0165.098.598.798.915:25:5698.998.999.199.199.0163.0165.098.598.799.015:26:2698.898.798.999.098.9164.0165.098.598.698.815:26:5698.998.899.099.098.9165.0165.098.598.798.915:27:2699.098.999.199.199.0165.0165.098.598.799.015:27:5698.898.798.999.098.9164.0165.098.598.698.915:28:2698.998.899.099.098.9164.0165.098.598.798.915:28:5699.199.099.299.299.1165.0166.098.598.899.015:29:2698.998.899.099.098.9165.0165.098.598.698.915:29:5698.998.899.099.098.9165.0165.098.698.799.015:30:2698.998.899.099.098.9164.0165.098.598.698.915:30:5698.998.899.099.098.9165.0165.098.698.799.015:31:2698.998.899.099.098.9165.0165.098.598.698.915:31:5698.998.899.099.098.9165.0165.098.598.798.915:32:2698.898.798.998.998.8164.0165.098.598.798.915:32:5698.998.899.099.098.9165.0165.098.698.798.915:33:2698.898.898.998.998.9165.0165.098.598.698.915:33:5698.998.899.099.098.9165.0165.098.698.798.915:34:2698.898.898.999.098.9164.0165.098.598.698.915:34:5698.998.899.099.098.9165.0165.098.698.799.015:35:2698.998.899.099.098.9164.0165.098.598.698.915:35:5698.998.899.099.098.9165.0165.098.698.799.015:36:2698.998.899.099.098.9165.0165.098.598.698.915:36:5698.998.899.099.098.9164.0165.098.698.799.015:37:2698.998.899.099.098.9165.0165.098.598.698.915:37:5699.098.999.199.099.0165.0165.098.598.799.115:38:2298.998.899.099.098.9165.0165.098.698.698.915:38:5692.892.893.193.493.0163.0163.094.392.492.615:39:5686.386.186.987.086.6161.0161.090.086.587.015:40:5680.580.380.580.980.6164.0164.082.880.179.615:41:5673.973.774.074.374.0162.0162.076.573.773.015:42:5667.967.867.868.167.9157.0158.070.967.967.915:43:5662.462.162.562.662.4162.0160.065.862.663.515:44:5658.057.958.258.158.1161.0160.061.358.059.115:45:5654.554.554.854.754.6161.0161.057.254.155.315:46:5651.451.351.451.451.4159.0159.053.650.952.315:47:5148.748.748.748.748.7156.0157.050.548.349.915:47:5648.548.448.648.548.5152.0152.050.248.049.5
15:48:5646.346.946.346.446.596.097.048.347.850.515:49:5644.945.345.044.945.057.056.048.547.949.515:50:5644.645.345.145.045.030.030.048.748.348.915:51:5645.645.545.445.445.514.014.048.748.748.815:52:1646.045.945.645.745.810.010.048.848.648.9由表2可以看出，2019s03批升温开始时间为14:52:36，灭活开始时间为15:08:22，冷却开始时间为15:38:22，冷却结束时间为15:47:51，排水、排汽结束时间为15:52:16。
34.表3：2019s04批干热灭活数据（℃、kpa）时间tatbtctd平均温度papbtlthte13:28:4623.823.724.124.023.90.00.024.624.223.913:28:5623.823.724.124.023.90.00.024.624.223.913:29:4630.630.730.130.230.44.04.028.630.429.613:30:4637.437.536.836.937.28.08.035.537.235.813:31:4643.944.043.443.543.713.013.042.143.842.413:32:4650.250.349.849.850.018.018.048.550.248.413:33:4655.855.855.655.655.724.024.054.355.954.413:34:4661.561.661.361.361.431.031.059.961.660.113:35:4667.467.467.167.167.339.039.065.767.565.813:36:4673.373.373.173.173.249.049.071.773.571.913:37:4679.279.279.179.079.161.060.077.679.577.913:38:4684.784.884.784.884.874.074.083.385.283.913:39:4689.489.389.689.689.587.087.088.489.788.713:40:4693.793.693.793.893.7101.0101.092.294.093.313:41:4697.297.197.297.297.2113.0113.096.097.596.813:42:4698.298.098.798.798.4161.0162.097.998.498.113:43:3898.498.398.898.898.6163.0163.098.298.498.313:44:1198.698.398.998.998.7164.0164.098.398.598.513:44:4198.698.498.998.998.7164.0164.098.498.598.613:45:1198.798.599.099.098.8164.0164.098.498.698.713:45:4198.798.599.199.198.9164.0164.098.598.798.813:46:1199.098.899.199.299.0165.0165.098.498.899.013:46:4198.998.798.998.998.9164.0165.098.498.998.613:47:1198.798.598.999.098.8164.0164.098.498.698.713:47:4198.798.699.099.098.8164.0164.098.598.698.813:48:1199.098.899.299.299.1165.0165.098.498.899.113:48:4198.798.598.998.998.8164.0164.098.498.698.713:49:1198.898.699.099.098.9164.0165.098.598.698.813:49:4199.198.999.199.299.1165.0165.098.498.899.013:50:1198.798.698.998.998.8164.0164.098.498.698.7
13:50:4198.898.799.099.098.9164.0165.098.598.698.813:51:1199.098.999.199.299.1165.0165.098.498.899.013:51:4198.798.699.099.098.8164.0164.098.498.698.713:52:1198.898.799.199.098.9165.0165.098.598.798.813:52:4198.898.798.998.998.8165.0165.098.498.898.713:53:1198.898.799.099.098.9164.0165.098.598.698.813:53:4199.299.199.199.299.1165.0166.098.598.999.013:54:1198.898.799.099.098.9164.0165.098.598.698.813:54:4199.299.099.199.299.1165.0165.098.598.999.113:55:1198.898.799.099.098.9164.0165.098.598.698.913:55:4199.199.099.199.299.1165.0166.098.598.899.113:56:1198.898.799.099.098.9165.0165.098.598.698.813:56:4199.299.199.099.199.1165.0166.098.498.999.013:57:1198.898.799.099.098.9165.0165.098.598.698.813:57:4199.199.099.199.299.1165.0166.098.598.899.113:58:1198.898.799.099.098.9164.0165.098.598.698.913:58:4199.299.199.199.199.1165.0166.098.598.999.113:59:1198.998.899.099.098.9164.0165.098.598.798.913:59:4199.198.999.099.099.0165.0166.098.599.098.914:00:1198.998.899.199.199.0165.0165.098.698.798.914:00:4198.898.799.099.098.9164.0165.098.598.698.914:01:1198.998.899.199.199.0164.0165.098.698.799.014:01:4198.898.899.099.098.9164.0165.098.598.698.914:02:1199.299.199.099.199.1166.0166.098.599.099.014:02:4198.998.899.099.199.0165.0165.098.698.798.914:03:1198.898.899.099.098.9164.0165.098.598.698.914:03:4199.098.999.199.199.0165.0165.098.698.799.014:04:1198.998.899.099.098.9165.0165.098.698.798.914:04:4198.898.798.998.998.8164.0165.098.598.698.814:05:1199.098.899.199.199.0165.0165.098.698.799.014:05:4198.998.899.099.098.9165.0165.098.598.698.914:06:1199.199.099.099.099.0165.0166.098.598.998.914:06:4198.998.899.199.199.0165.0165.098.698.799.014:07:1198.998.899.099.098.9165.0165.098.598.698.914:07:4199.199.098.999.099.0165.0166.098.598.999.014:08:1198.998.899.199.199.0165.0165.098.698.799.014:08:4198.998.899.099.098.9164.0165.098.598.698.914:09:1199.199.099.099.099.0165.0166.098.598.998.914:09:4198.998.899.099.098.9165.0165.098.698.798.9
14:10:1198.998.899.099.098.9164.0165.098.598.698.914:10:4199.199.099.199.299.1165.0166.098.598.899.114:11:1198.998.899.099.098.9165.0165.098.698.798.914:11:4198.998.899.099.098.9164.0165.098.598.698.914:12:1199.299.199.199.199.1165.0166.098.598.899.114:12:4198.998.899.099.098.9165.0165.098.698.798.914:13:1198.998.899.099.098.9165.0165.098.598.698.914:13:3899.199.099.099.099.0165.0166.098.598.999.014:14:1192.592.492.592.592.5163.0163.094.292.192.214:15:1185.885.787.787.586.7161.0161.089.586.186.414:16:1180.280.079.579.479.8165.0166.082.279.779.414:17:1173.373.473.072.973.2159.0159.075.873.172.914:18:1167.267.067.667.467.3158.0160.070.067.167.114:19:1161.261.362.462.461.8159.0160.064.761.662.314:20:1156.556.558.057.957.2158.0158.059.956.858.214:21:1152.852.754.154.053.4158.0158.055.752.454.414:22:1149.449.450.550.449.9159.0160.051.848.950.514:22:1849.149.149.749.849.4158.0160.051.149.049.814:23:1147.948.648.348.648.4105.0105.050.049.751.314:24:1145.946.846.346.846.562.061.050.149.650.514:25:1145.346.246.246.346.032.032.050.249.849.914:26:1145.846.946.747.146.614.014.050.249.949.614:26:3046.147.046.847.446.810.010.050.350.149.6由表3可以看出2019s04批升温开始时间为13:28:56，灭活开始时间为13:43:38，冷却开始时间为14:13:38，冷却结束时间为14:22:18，排水、排汽结束时间为14:26:30。
35.由表1-3可以看出，2019s02、2019s03、2019s04三批升温时间分别为856s、946s、882s，冷却时间分别为571s、569s、520s，工序总时间分别为3492s、3590s、3464s，结合图2-4，升温过程平滑快速均匀，灭活过程温度稳定，冷却过程快速。
36.三个批次干热灭活前后效价对比如下表4。
37.表4：2019s02批、2019s03批、2019s04批干热前后效价（iu/瓶）由表4可以看出2019s02批、2019s03批、2019s04批干热前后效价损失分别为8.86%、7.91%、4.34%，相比较其他因子类产品干热灭活效价损失15%-30%，采用本工艺人凝血因子ix的效价损失小。
38.实施例3按照实施例2的干热灭活工艺对人凝血因子
ⅸ
中猪细小病毒（ppv）和脑心肌炎病毒（emcv）的灭活效果进行验证。
39.将2014s06、2014s07、2014s08三批样品分别按照9:1加入猪细小病毒、脑心肌炎病毒，充分混匀。将样品放入冻干机后，进行冻干，冻干前、冻干后及加入病毒后零时对照取样后立即-70℃冻存。处理样品放入干热灭活柜中平衡，当水温上升至98.2℃时开始计时，进行98.2-99.2℃恒温加热处理，分别于5、15、30分钟取样。在96孔细胞培养板中检测其病毒滴度。每批样品至少重复测定二次。
40.实验结果参见表5-6及图5-6。
41.表5：干热法灭活后人凝血因子
ⅸ
样品中残余ppv滴度（lgtcid
50
/0.1ml）表6：干热法灭活后人凝血因子
ⅸ
样品中残余emcv滴度（lgtcid
50
/0.1ml）《血液制品去除灭活病毒技术方法及验证指导原则》规定经过病毒挑战性试验后，病毒降低量≥4lgtcid
50
/0.1ml可认定灭活病毒方法有效。本次病毒灭活可使病毒降低量≥4lgtcid
50
/0.1ml，灭活方法有效。
42.尽管通过参考附图并结合优选实施例的方式对本发明进行了详细描述，但本发明并不限于此。在不脱离本发明的精神和实质的前提下，本领域普通技术人员可以对本发明的实施例进行各种等效的修改或替换，而这些修改或替换都应在本发明的涵盖范围内/任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，可轻易想到变化或替换，都应涵盖在本发明的保护范围之内。

技术特征：
1.一种凝血因子
ⅸ
的干热灭活装置，包括灭活内室，灭活内室分别连通压缩空气管线、排汽管线、注水管线和排泄管线，压缩空气管线设有压缩空气阀，排汽管线设有排汽阀，注水管线设有注水阀，排泄管线设有排泄阀；其特征在于，灭活内室顶部和底部分别设有温度传感器，灭活内室顶部还设有压力传感器；灭活内室顶部通过换热器与蒸汽管线、冷却水管线连通，灭活内室顶部与换热器间的连通管线上设有温度传感器，蒸汽管线包括第一蒸汽子管线和第二蒸汽子管线，第一蒸汽子管线设有大蒸汽阀，第二蒸汽子管线设有小蒸汽阀，冷却水管线包括第一冷却水子管线和第二冷却水子管线，第一冷却水子管线设有大冷却水阀，第二冷却水子管线设有小冷却水阀；灭活内室底部通过换热器与排水管线、疏水管线和排冷凝水管线连通，排水管线设有排冷却水阀，疏水管线设有疏水阀，排冷凝水管线设有冷凝水旁通阀，灭活内室底部与换热器间的连通管线上设有温度传感器。2.如权利要求1所述的干热灭活装置，其特征在于，灭活室内底部设置至少两个温度传感器，灭活室内顶部设置至少两个压力传感器。3.如权利要求1所述的干热灭活装置，其特征在于，小蒸汽阀为比例调节阀。4.如权利要求1所述的干热灭活装置，其特征在于，灭活室底部与换热器的连通管线上还设有循环泵。5.如权利要求1所述的干热灭活装置，其特征在于，干热灭活装置还包括控制器，温度传感器、压力传感器、大蒸汽阀、小蒸汽阀、大冷却水阀、小冷却水阀、压缩空气阀、排汽阀、注水阀、排泄阀、排冷却水阀、疏水阀、冷凝水旁通阀分别与控制器电性连接，控制器还与显示器电性连接。6.一种人凝血因子
ⅸ
的干热灭活工艺，其特征在于，采用如权利要求1-5任一所述的干热灭活装置，步骤如下：s1注水；s2升温：开始升温时大蒸汽阀间歇打开，小蒸汽阀开度调节至100%，升温速度为6.0-7.5℃/s，当灭活内室底部的温度传感器温度到达95.0℃后大蒸汽阀关闭，当灭活内室底部的温度传感器温度到达97.0℃后，小蒸汽阀开度调节至80%，当灭活内室底部的温度传感器温度到达97.5℃时通压缩空气，使灭活内室压力到达160kpa并维持压力在160-165kpa，疏水阀、冷凝水旁通阀开启；s3灭活：当灭活内室底部的温度传感器温度到达98.2℃，小蒸汽阀开度调节至50%，当灭活内室底部的温度传感器温度到达98.6℃，小蒸汽阀开度调节至15%且间歇开启，使灭活温度保持在98.2-99.2℃，间歇通入压缩空气使灭活内室压力保持在160-165kpa，灭活时间为1800s；s4冷却：疏水阀、冷凝水旁通阀关闭，间歇通入压缩空气使灭活内室压力保持在155-160kpa，小冷却水阀打开30s、关闭30s，当灭活内室底部的温度传感器温度到达90.0℃后，排冷却水阀打开，大冷却水阀与小冷却水阀同时打开30s、关闭30s，当灭活内室底部的温度传感器温度低于50.0℃冷却结束；s5排水、排汽。7.如权利要求6所述的干热灭活工艺，其特征在于，s1注水步骤为：打开注水阀，注入去离子水，去离子水的液位控制在150-300mm之间，去离子水的电导率小于60us/cm。
8.如权利要求6所述的干热灭活工艺，其特征在于，干热灭活工艺的处理量为2000-6000瓶/批。9.如权利要求6所述的干热灭活工艺，其特征在于，s2升温步骤小于1000s，s4冷却步骤小于600s。10.如权利要求6所述的干热灭活工艺，其特征在于，s5排水、排汽步骤为：排汽阀打开，排泄阀打开，灭活内室顶部压力传感器的压力低于10kpa时排水、排汽结束。

技术总结
本发明涉及干热灭活技术领域，具体涉及一种人凝血因子


技术研发人员：苏登城 李杰 李昌成 陈勇 褚风磊
受保护的技术使用者：山东泰邦生物制品有限公司
技术研发日：2023.07.07
技术公布日：2023/10/15