Sudjarwo Sri Agus, Eraiko Koerniasari, Sudjarwo Giftania Wardani
Department of Pharmacology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia.
Department of Conservative Dentistry, Faculty of Dentistry, Airlangga University, Surabaya 60115, Indonesia.
J Adv Pharm Technol Res. 2017 Jul-Sep;8(3):91-96. doi: 10.4103/japtr.JAPTR_167_16.
The aim of this study was to characterize of chicken egg yolk immunoglobulins (IgYs) specific as immunotherapy to complex (MTBC) infection. Lohmann laying hens were immunized intramuscularly with antigenic of MTBC. Egg yolk was separated from egg white, and IgY antibody was then purified by multiple polyethylene glycols 6000 extraction and ammonium sulfate purification steps. The IgY anti-MTBC concentration in egg yolk increased at 2 weeks and it reached a maximum at 4 weeks after immunization. After 6 weeks, the levels of IgY anti-MTBC decreased gradually. The antibody of MTBC was detected and produces a specific line of precipitation in agar gel precipitation test beginning the week 2 after the first immunization. Analysis of results obtained with ELISA showed a significant increase in the MTBC specific antibodies after 2 weeks and reached a plateau at 4 weeks from the booster immunization. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the IgY preparation to be pure and dissociated into protein bands with molecular weights of 112, 78, 69, 49, and 28 kDa and Western blot analysis shown the presence of anti-MTBC IgY in egg yolks, with molecular weights of approximately 78 kDa. These results suggested that egg yolk could be a practical strategy in large-scale production of specific anti-MTBC IgY for immunotherapy of TBC.
本研究的目的是鉴定作为免疫疗法用于结核分枝杆菌复合群（MTBC）感染的鸡卵黄免疫球蛋白（IgY）。用MTBC抗原对罗曼蛋鸡进行肌肉注射免疫。将蛋黄与蛋清分离，然后通过多次聚乙二醇6000提取和硫酸铵纯化步骤纯化IgY抗体。蛋黄中IgY抗MTBC的浓度在免疫后2周增加，并在4周时达到最高。6周后，IgY抗MTBC的水平逐渐下降。在首次免疫后第2周开始的琼脂凝胶沉淀试验中检测到MTBC抗体并产生特异性沉淀线。ELISA结果分析显示，2周后MTBC特异性抗体显著增加，并在加强免疫后4周达到平台期。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示IgY制剂纯净，解离为分子量为112、78、69、49和28 kDa的蛋白条带，蛋白质印迹分析显示蛋黄中存在抗MTBC IgY，分子量约为78 kDa。这些结果表明，蛋黄可能是大规模生产用于结核病免疫治疗的特异性抗MTBC IgY的一种实用策略。
