Aavula Shukra M, Nimmagadda Sridevi V, Biradhar Neelakantam, Sula Samuel, Chandran Dev, Lingala Rajendra, Villuppanoor Srinivasan Alwar
Research and Development Center, Indian Immunologicals Limited, Rakshapuram, Gachibowli, Hyderabad 500032, India.
Biotechnol Res Int. 2011;2011:652147. doi: 10.4061/2011/652147. Epub 2011 Oct 5.
Recombinant antibody phage display technology is a vital tool that facilitates identification of specific binding molecules to a target enabling the rapid generation and selection of high affinity, fully human, or mouse antibody product candidates essentially directed towards disease target appropriate for antibody therapy. In this study, a recombinant single-chain Fv antibody fragment (scFv) A11 was isolated from immune spleen cells obtained from mice immunized with inactivated rabies virus (Pasteur strain) using standard methodology and was characterized for its specificity towards the rabies virus glycoprotein. Epitope mapping using peptide libraries and truncated glycoprotein polypeptides suggested that A11 bound to the antigenic site II of rabies glycoprotein against which a majority of rabies virus neutralizing antibodies are directed. The use of the above technology could, therefore, allow development of scFvs with different specificities against the rabies glycoprotein as an alternative to the more cumbersome protocols used for the development of monoclonal antibodies.
重组抗体噬菌体展示技术是一种重要工具,有助于鉴定与靶标特异性结合的分子,从而能够快速生成并筛选出高亲和力、全人源或小鼠源抗体候选产品,这些产品主要针对适合抗体治疗的疾病靶标。在本研究中,使用标准方法从用灭活狂犬病病毒(巴斯德株)免疫的小鼠的免疫脾细胞中分离出重组单链Fv抗体片段(scFv)A11,并对其针对狂犬病病毒糖蛋白的特异性进行了表征。使用肽库和截短的糖蛋白多肽进行表位作图表明,A11与狂犬病糖蛋白的抗原位点II结合,大多数狂犬病病毒中和抗体都针对该位点。因此,使用上述技术可以开发出针对狂犬病糖蛋白具有不同特异性的scFv,作为开发单克隆抗体所使用的更为繁琐方案的替代方法。