Chappel J A, He M, Kang A S
Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
J Immunol Methods. 1998 Dec 1;221(1-2):25-34. doi: 10.1016/s0022-1759(98)00094-5.
Here we describe a phage vector for the display of single chain antibodies and polypeptides on the surface of filamentous M13 phage which permits facile manipulation of the valency of display. The gene encoding the polypeptide is fused to a synthetic copy of the major coat protein VIII gene (gpVIII) which permits incorporation into the phage during assembly of the filament. Here we examine the growth parameters of phage propagation on the subsequent selection of an anti-progesterone antibody fragment from a mixture of display phage. Our results suggest that the density of the polypeptides displayed on phage may be modulated by altering growth conditions. This ability to influence polypeptide display density on filamentous phage may provide a versatile approach for accessing complex libraries and the capture of weaker ligand receptor interactions by avidity, whilst the potential to access and discriminate between higher affinity interactions is not negated.
在此，我们描述了一种用于在丝状M13噬菌体表面展示单链抗体和多肽的噬菌体载体，它能够方便地操控展示的价态。编码多肽的基因与主要外壳蛋白VIII基因（gpVIII）的合成拷贝融合，这使得在丝状噬菌体组装过程中能够掺入噬菌体。在此，我们研究了噬菌体增殖的生长参数，以及随后从展示噬菌体混合物中筛选抗孕酮抗体片段的情况。我们的结果表明，通过改变生长条件可以调节噬菌体上展示的多肽密度。这种影响丝状噬菌体上多肽展示密度的能力，可能为筛选复杂文库以及通过亲和力捕获较弱的配体-受体相互作用提供一种通用方法，同时并不排除获取和区分更高亲和力相互作用的可能性。
