PREPARATION AND CHARACTERIZATION OF POLYCLONAL ANTI-RPRESSS OF HBSAG

HBSAG 多克隆抗抑制物的制备和表征

• 批准号: 3804908
• 负责人: M W YU
• 金额: --
• 依托单位: CENTER BIOLOGICS EVALUATION RESEARCH HEMATOLOGY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3804908
• 关键词: Freund's adjuvant; antibody; hemocyanin; hepatitis B antigens; laboratory rabbit; peptides; radioimmunoassay; synthetic peptide; western blottings

Two rabbits were immunized with multiple injections of recombinant preS1 peptide (rpreS1) utilizing the commercial Ribi Adjuvant System. Sera were screened for specific anti-rpreS1 by immune blotting and by radioimmunoassay; very little specific antibody was detected. Two additional rabbits were then immunized with the same immunogen in Freund's Adjuvant, and although a good response was observed, the antisera were not specific for preS1, suggesting that the rpreS1 immunogen was not pure. The anti-preS1 titers observed in these sera were lower than those of a previous preparation obtained by immunization with a synthetic preS1 peptide (residues 21-47) conjugated to keyhole limpet hemocyanin, and they were also lower than that of commercially available monoclonal anti-preS1. Because the rpreS1 preparation was recently found to contain two peptides, separable on 10% Tricine gels, we have now immunized two rabbits with the material from these separated bands. It is hoped that this will yield specific antibodies against preS1.

用重组preS 1多次注射免疫两只家兔 肽（rpreS 1），利用商业Ribi佐剂系统。 血清 通过免疫印迹法和免疫荧光法筛选特异性抗rpreS 1抗体。 放射免疫测定法;检测到非常少的特异性抗体。 两 然后用相同的免疫原免疫另外的兔子， 弗氏佐剂，虽然观察到良好的反应， 抗血清对preS 1无特异性，提示rpreS 1 免疫原不纯。 在这些血清中观察到的抗preS 1滴度 低于通过免疫获得的先前制剂的那些 用合成的preS 1肽（残基21-47）与钥匙孔结合 血蓝蛋白，也低于市售 可获得单克隆抗preS 1。 因为rpreS 1制备是 最近发现含有两种肽，可在10% Tricine凝胶上分离， 我们现在已经用这些分离的材料免疫了两只兔子， 乐队. 希望这将产生特异性抗体， preS1。