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Fluobody Production

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Creative Biolabs offers custom fluobody construction service for a series of research, diagnostic, and therapeutic applications. Compared to the traditional chemical linked organic fluorochromes, the fluobody holds more advantages, such as strong stability and high sensitivity. Creative Biolabs has established an advanced fluobody construction platform which could facilitate the success of our customers’ projects in an efficient and cost-effective manner.

Nowadays, immunofluorescence (IF) is significant in flow cytometry for both localization studies and diseases diagnosis. Conventionally, the antibodies are often chemically conjugated with the fluorochrome, such as fluorescein isothiocyanate (FITC). However, such conjugation is labile with illumination, and the fluorochrome may be conjugated to the antigen binding sites which affect the antigen binding capacity of the antibodies. To overcome these drawbacks, the construction of fluobody has become a substitute to chemical antibody-fluorochrome conjugation. Compared to the conventional chemical conjugation format, fluobody has comparable but more stable excitation and emission spectra without influence on the antigen binding capacity, which is more suitable for the immunofluorescence (IF) applications. At present, Creative Biolabs is able to provide such fluobody construction service by fusing the interested antibody with various fluorescent proteins (e.g. GFP) to meet our clients’ specific demands.

Green fluorescent protein (GFP) is a 238 amino acids protein discovered in jellyfish. GFP is intensely fluorescent and stable in a variety of conditions. It has the same emission wavelength with FITC at 510 nm. The wild-type GFP has a major excitation peak at 395 nm and a minor excitation peak at 470 nm. In addition, a large number of GFP variants have been developed till now. They hold differences in their spectral characteristics, fluorescence intensity, stability, and other properties which enlarge the toolbox of the fluorescent protein labeling.

Instead of chemical coupling, Creative Biolabs has developed a fluobody construction platform which genetically fusing the antibody and GFP encoding genes in a 1:1 ratio. The combination of antibody and fluorochrome could enhance the accuracy in quantitative control. Furthermore, the fused GFP is relatively more stable and not easy to be inactivated. We are able to clone and express recombinant antibodies in E. coli and a variety of other heterologous production platforms.

Figure. 1 Genetic fusion of the GFP-Fluobody. (Kim et al. 2002) Figure. 1 Genetic fusion of the GFP-Fluobody. (Kim et al. 2002)

With our advanced phage display platform and hybridoma technology, Creative Biolabs is able to select single-chain variable fragment (scFv), which contains the variable regions of the heavy and light chain connected by a short linker sequence. The antibody genes will be cloned and fused with the GFP encoding gene in the optimal expression system. The constructed fluobodies could be applied to detect a variety of targets, such as plant pathogen and picloram herbicide. In addition to the GFP, our scientists can also generate custom fluobodies with many other fluorescent protein variants.

Table 1. Properties of the best FP variants. (Shaner et al. 2005)

Table 1. Properties of the best FP variants. (Shaner et al. 2005)

Creative Biolabs is a leading service provider in the field of antibody research and development. We are proud to provide the custom fluobody service for immunofluorescence. Our Ph.D. level scientists are pleased to offer the best solutions and the high-quality products tailored to our clients’ requirements.

Reference:
Kim I S, Shim J H, Suh Y T, et al. Green fluorescent protein-labeled recombinant fluobody for detecting the picloram herbicide[J]. Bioscience, biotechnology, and biochemistry, 2002, 66(5): 1148-1151.
Shaner N C, Steinbach P A, Tsien R Y. A guide to choosing fluorescent proteins[J]. Nature methods, 2005, 2(12): 905-909.




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