Creative Biolabs provides the first-class GFP scaffold library construction service for customers. With advanced library construction platform and years of experience in scaffold library construction, our top scientists have built about 55 kinds of scaffold libraries for clients all over the world.
Green fluorescent protein (GFP) is a highly stable molecule characterized by its high quantum yield of fluorescence which depends on the fluorochrome linked in the core. GFP has shown a unique “β-can” formation through structure analysis. It is formed by an extremely well packed β barrel, which is a nearly perfect cylinder, and five shorter α helices on the ends of the structure. In addition, the β barrel consists of 11 β-stranded with a fully buried central α helix that contains a chromophore [4-(p-hydroxybenzylidene)-imidazolidin-5-one (HBI), shown in green]. The fluorescence is generated in an autocatalytic chemical reaction of neighboring residue interactions. Through appropriate modification of these residues, it is available to produce a large variety of GFP derivatives for different characteristics and applications.
Meanwhile, GFP can be developed as a kind of autofluorescent scaffolds and applied as probes for bio-imaging purposes. It is generally difficult to measure the quality and characteristics of phage libraries displayed in vivo in a quantitative manner. Whereas, with an autofluorescent scaffold (e.g. GFP), the properties of the library can be easily quantified. In addition, the expression level and bias of the generated library can also be quantified by a flow sorter (e.g. a FACS machine). This method allows an intracellular screening and enrichment for library sequences with predetermined characteristics, e.g. selection of high expression of correctly folded protein by identifying the clones with high fluorescence. According to the advanced technology, Creative Biolabs is pleased to offer GFP scaffold libraries with high diversity to facilitate our customers’ research and project development.
The GFP scaffold libraries are generated through Hi-Affi™ phage display platform, a proprietary technology of Creative Biolabs. This platform is an improved phage display system using the trimer codon technology and NNK method. Compared with the traditional phage display, Hi-Affi™ platform is more suitable for screening and selecting of high affinity target binders. Ultimately, the HiAffi™ platform can offer 100% precise mutant library construction with over 1010 diversity.
With extensive experience in the field of GFP scaffold library construction, Creative Biolabs is persistent in providing high quality products, excellent credit and passionate service for our global customers.
Fig. 1 S65T Q80R Green Fluorescent Protein (GFP) pH 8.5. (PDB ID: 1Q4A)