Development of process to produce polyvalent IgY antibodies anti-African snake venom
Development of process to produce polyvalent IgY antibodies anti-African snake venom
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DOI：
10.1016/j.toxicon.2008.05.022
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发表时间：
2008-08-01
期刊：
影响因子：
2.8
通讯作者：
da Silva, Wilmar Dias
中科院分区：
文献类型：
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作者：
Costa de Almeida, Claudia Maria;da Silva, Claudia Leticia;da Silva, Wilmar Dias
用B制备多价抗Bitis和抗眼镜蛇蛇毒血清IgY抗体。arietans，B. nasicornis，B. B. rhinoceros，新几内亚线虫N. melanoleuca和N. mossambica毒液来免疫鸡。在10个月的免疫期之前和期间收集血液和鸡蛋;然后制备血清和蛋黄提取物，并通过ELISA和Western印迹法测定抗蛇毒血清抗体的存在。ELISA抗蛇毒血清抗体滴度，称为U-ELISA/ml血清或蛋黄提取物，在免疫前血清或蛋黄中不存在，在免疫后4周期间急剧增加，此后达到平台。用ELISA检测到高抗蛇毒效价的卵黄提取物分离和纯化IgY。纯化的IgY制剂识别从10至20 kDa至60和70 kDa的毒液蛋白条带，如Western印迹所示。抗蛇毒血清的回收率在80%以上。最终的制备物显示出高的抗蛇毒活性（> 100，000 U-ELISA/ml）以及中和毒液致死性的效力（1440 μ g IgY中和62.2LD 50的毒液），并且没有毒性产物、热原或细菌和真菌污染。(C)2008爱思唯尔有限公司保留所有权利。
Polyvalent anti-Bitis and anti-Naja antivenom IgY antibodies were prepared using B. arietans, B. nasicornis, B. rhinoceros, N. melanoleuca, and N. mossambica venoms to immunize chickens. Blood and eggs were collected before and during the 10-month immunization period; the sera and yolk extracts were then prepared and assayed for the presence of antivenom antibodies by ELISA and Western blot methods. ELISA Antivenom antibody titers, referred to as U-ELISA/ml of serum or egg yolk extracts, absent in pre-immunization sera or yolk, increased sharply during the 4 weeks after immunization, reaching a plateau thereafter. Yolk extracts with high antivenom titers, as detected by ELISA were used to isolate and purify IgY. Purified IgY preparations recognized venom protein bands from 10 to 20 kDa to 60 and 70 kDa, as shown by Western blot. Recovery of antivenom antibodies from the whole yolk was over 80%. Final preparations exhibited high antivenom activity (> 100,000 U-ELISA/ml) as well as efficacy in neutralizing venom lethality (1440 mu g of IgY neutralize 62.2 LD50 of venom), and were free of toxic products, pyrogen or bacterial and fungal contaminations. (C) 2008 Elsevier Ltd. All rights reserved.