释义：

Low-affinity interaction detection method designed specifically to detect interactions between extracellular proteins. Recombinant soluble fragments of these proteins are produced in a (generally) mammalian expression cell line and secreted into the medium and produced in two forms: a biotinylated bait which can be captured on a streptavidin-coated solid phase suitable for screening, and a pentamerised enzyme-tagged (beta-lactamase) prey. The bait and prey proteins are presented to each other in a binary fashion to detect direct interactions between them, similar to a conventional ELISA. The pentamerisation of the proteins in the prey is achieved through a peptide sequence from the cartilage oligomeric matrix protein (COMP) and increases the local concentration of the ectodomains thereby providing significant avidity gains to enable even very transient interactions to be detected. By normalising the activities of both the bait and prey to predetermined levels prior to screening, interactions having monomeric half-lives of 0.1 sec can be detected with low false positive rates.

一种低亲和力相互作用检测方法，专门用于检测细胞外蛋白质之间的相互作用。这些蛋白质的重组可溶性片段在 (通常是) 哺乳动物表达细胞系中生产，并分泌到培养基中，生产两种形式：一种是生物素化的诱饵，可被捕获在适合筛选的链霉亲和素涂层固相上，另一种是五聚体酶标记 (β-内酰胺酶) 的猎物。诱饵和猎物蛋白以二元方式呈现给彼此，以检测它们之间的直接相互作用，类似于传统的 ELISA。猎物中蛋白质的五聚体化通过来自软骨寡聚基质蛋白 (COMP) 的肽序列实现，增加了外显域的局部浓度，从而提供显著的亲和力增益，使得即使是非常短暂的相互作用也能被检测到。通过在筛选前将诱饵和猎物的活性归一化到预定的水平，可以以低假阳性率检测到具有 0.1 秒单体半衰期的相互作用。