Van Wyngaardt W, Du Plessis D H
Immunology Division, Onderstepoort Veterinary Institute, South Africa.
Onderstepoort J Vet Res. 1998 Jun;65(2):125-31.
A filamentous phage library displaying a vast repertoire of synthetic single chain fragment variable (scFv) antibody fragments was subjected to affinity selection on purified bluetongue virus (BTV) particles. After four rounds of selection and amplification, 73 out of a total of 90 fusion phage clones tested were found to bind to purified BTV in ELISA. One of these, the clone producing the highest ELISA signal, was selected for an investigation of its potential as an immunodiagnostic reagent. The binding of this phage antibody (designated A12) could be inhibited by free virus and by antibodies in immune serum. Inhibition with antibodies in guinea-pig sera suggested that it recognized an antigenic region on BTV that was similar on at least 10 different BTV serotypes. A sandwich ELISA utilizing antibody A12 was capable of detecting approximately 60 ng of purified BTV.
一个展示大量合成单链抗体可变区（scFv）抗体片段的丝状噬菌体文库，在纯化的蓝舌病毒（BTV）颗粒上进行了亲和筛选。经过四轮筛选和扩增后，在总共测试的90个融合噬菌体克隆中，有73个被发现在酶联免疫吸附测定（ELISA）中能与纯化的BTV结合。其中一个产生最高ELISA信号的克隆被选来研究其作为免疫诊断试剂的潜力。这种噬菌体抗体（命名为A12）的结合可被游离病毒和免疫血清中的抗体抑制。用豚鼠血清中的抗体进行抑制表明，它识别BTV上至少10种不同BTV血清型中相似的一个抗原区域。利用抗体A12的夹心ELISA能够检测到约60 ng的纯化BTV。
