Ruiz Díaz Nancy, Cisternas Carlos, Silva Mauricio, Hernández Adrián, Chacana Pablo
Programa de Doctorado en Ciencias Agropecuarias, Facultad de Recursos Naturales, Universidad Católica de Temuco, Temuco, Chile.
Escuela de Medicina Veterinaria, Facultad de Recursos Naturales y Medicina Veterinaria, Universidad Santo Tomás, Temuco, Chile.
Front Vet Sci. 2024 Apr 12;11:1382510. doi: 10.3389/fvets.2024.1382510. eCollection 2024.
Anti-soybean agglutinin (SBA) IgY was produced, and its potential to neutralize the haemagglutinating activity of SBA was tested. Thirty-five-week-old hens [treatment ( = 5) and control ( = 5)] were immunized with SBA or injected with saline 4 times every 15 days. Eggs were collected after the last immunization, and IgY was extracted using the polyethylene glycol (PEG) method. Serum anti-SBA IgY titres in immunized hens increased after the first immunization and reached a plateau between days 45 and 60. In contrast, specific IgY titres in the control group remained at basal levels throughout the evaluation. Average IgY titres were significantly higher in the treatment group on days 15, 30, 45, and 60. Total IgY content in the egg yolk extract was 38.7 ± 1.6 and 37.7 ± 1.5 mg/ml for the treatment and control groups, respectively. The specific anti-SBA IgY titer detected in the egg yolk extract was significantly higher ( < 0.001) for hens in the treatment group compared to the control group, with OD values of 0.98 ± 0.05 and 0.058 ± 0.02, respectively. The specificity of anti-SBA IgY was confirmed by the Western blotting, and the inhibition of SBA-induced haemagglutination was compared with D-galactose, a known molecule that binds to SBA and blocks its binding to erythrocytes. The inhibition of SBA-induced haemagglutination by the anti-SBA IgY reached 512 units of haemagglutination inhibition (UHI), compared to 8 or 256 UHI, respectively, when IgY from control chickens or D-galactose was used. Thus, anti-SBA IgY antibodies were efficiently produced in large quantities and effectively inhibited SBA-induced haemagglutination .
制备了抗大豆凝集素(SBA)IgY,并测试了其中和SBA血凝活性的潜力。对35周龄的母鸡[处理组(n = 5)和对照组(n = 5)]每15天用SBA免疫或注射生理盐水4次。最后一次免疫后收集鸡蛋,并用聚乙二醇(PEG)法提取IgY。免疫母鸡的血清抗SBA IgY滴度在首次免疫后升高,并在第45天至60天之间达到平台期。相比之下,对照组的特异性IgY滴度在整个评估过程中保持在基础水平。在第15、30、45和60天,处理组的平均IgY滴度显著更高。处理组和对照组蛋黄提取物中的总IgY含量分别为38.7±1.6和37.7±1.5 mg/ml。处理组母鸡蛋黄提取物中检测到的特异性抗SBA IgY滴度显著高于对照组(P < 0.001),OD值分别为0.98±0.05和0.058±0.02。通过蛋白质印迹法确认了抗SBA IgY的特异性,并将抗SBA IgY对SBA诱导的血凝的抑制作用与D-半乳糖进行了比较,D-半乳糖是一种已知的与SBA结合并阻断其与红细胞结合的分子。抗SBA IgY对SBA诱导的血凝的抑制作用达到512个血凝抑制单位(UHI),而使用对照鸡的IgY或D-半乳糖时分别为8或256 UHI。因此,抗SBA IgY抗体被大量高效地产生,并有效抑制了SBA诱导的血凝。