Chiurciu Constantin, Chiurciu Viorica, Oporanu Mariana, Pătrașcu Ionel Victor, Mihai Iuliana, Tablică Mădălina, Cristina Romeo Teodor
S.C. Romvac Company S.A., Soseaua Centurii 7, Voluntari, 77190 Ilfov, Romania.
Department of Pharmacology and Pharmacy, Faculty of Veterinary Medicine, Banat's University of Agriculture and Veterinary Medicine "King Michael I of Romania" from Timișoara, 119 Calea Aradului, 300645 Timișoara, Romania.
Evid Based Complement Alternat Med. 2017;2017:8671271. doi: 10.1155/2017/8671271. Epub 2017 Mar 20.
Characterization and evaluation of immunotherapeutic potential of ovotransferrin PC2 (OTf PC2) were performed in this study. The ovoprotein was obtained from egg white from hens immunized with bacterial antigens, pathogenic for humans. For the negative control samples, OTf was extracted from eggs collected from Specific Pathogen-Free (SPF) hens and purified by affinity chromatography on Protein G-agarose column with two eluting peaks: I, representing ovalbumin, and II, ovotransferrin. The final OTf form was reached by successive precipitation with ammonium sulfate and citric acid and the OTf form by saturating the form with FeCl. Multiple OTf PC2 samples were analyzed through Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and, based on the molecular marker migration model, the ovotransferrin (76.5 kDa) and ovalbumin (45 kDa) were detected. The agglutination reaction exhibited statistically significant high specificity of the multiple OTf PC2, by reacting with the antigens used for hens' immunization. Following ELISA, it was established that OTf PC2 from hyperimmune eggs has specificity for all antigens; the antibody titer was high, indicating that OTf PC2 possesses immunological properties similar to immunoglobulin Y (Ig Y). This study suggests that OTf PC2 immunological activity may play a crucial role in the prevention and treatment of infections resistant to antibiotics and OTf PC2 can also act as a valuable nutraceutical.
本研究对卵转铁蛋白PC2(OTf PC2)的免疫治疗潜力进行了表征和评估。该卵蛋白从用对人类致病的细菌抗原免疫的母鸡的蛋清中获得。对于阴性对照样品,OTf从无特定病原体(SPF)母鸡所产鸡蛋中提取,并通过在蛋白G - 琼脂糖柱上进行亲和层析纯化,有两个洗脱峰:峰I代表卵清蛋白,峰II代表卵转铁蛋白。最终的OTf形式通过硫酸铵和柠檬酸连续沉淀获得,而OTf形式通过用FeCl饱和该形式获得。多个OTf PC2样品通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)进行分析,基于分子标记迁移模型,检测到了卵转铁蛋白(76.5 kDa)和卵清蛋白(45 kDa)。凝集反应显示多个OTf PC2具有统计学上显著的高特异性,可与用于母鸡免疫的抗原发生反应。通过酶联免疫吸附测定(ELISA)确定,来自高免疫鸡蛋的OTf PC2对所有抗原均具有特异性;抗体滴度很高,表明OTf PC2具有与免疫球蛋白Y(IgY)相似的免疫学特性。本研究表明,OTf PC2的免疫活性可能在预防和治疗对抗生素耐药的感染中起关键作用,并且OTf PC2还可作为一种有价值的营养保健品。