Half-life Assay by Fluorescent
For Research Use Only. Not for Clinical Use.
Fig.1 Sketch of half-life assay methods by fluorescent. (Eldeeb, 2019)
It is well known that most therapeutic proteins suffer from poor pharmaceutical properties due to endogenous degradation, hence it is important to acquire precise kinetic spatiotemporal data regarding rates of degradation in diverse kinds of mammalian cells. Many fluorescent protein-based tools have been developed to address this experimental issue. As a leading biotechnology service provider, Creative Biolabs has established a mature platform for protein expression and fluorescence modification. Based on rich experience and proficient skills in protein engineering, we offer rapid and inexpensive half-life assay service by fluorescent to support your protein drugs research.
Introduction of Half-life Assay by Fluorescent
Fluorescent protein research began from the discovery of the green fluorescent protein (GFP). In recent years, a variety of multifunctional fluorescent proteins were expressed as fluorescent labels that can be incorporated into proteins by genetic fusion. Among them, fluorescent proteins with features of photo-activation, photo-switching, or photo-converting are popularly used in the proteins half-life assay in vitro owe to their many advantages such as low immunogenicity, high stability, and strong fluorescence. Moreover, the introduction of kinds of red fluorescent proteins enables multicolor labeling, further promotes novel fluorescent-based half-life assay technology development. Nowadays, fluorescent-based half-life assay has gradually become the most applied method to measure the pharmacokinetic properties of recombinant proteins because it is suitable for almost all proteins and various cell lines. Therefore, therapeutic proteins' half-life measured by fluorescence in vitro is also often regarded as an indicator of marketing potential.
Applications of Half-life Assay by Fluorescent
- Fluorescent switcher
Photo-switchable fluorescent proteins are exploited in proteins dynamics measurement for those fluorescent protein fragments that can acquire or lose fluorescence with intervention. Following distinct population of labeled proteins is produced using specific wavelength and intensity of light, the speed with which unlabeled proteins replace tagged ones is quantified and the half-life of the protein can be estimated.
- Fluorescent timers
A novel approach to monitoring protein degradation rate is utilizing fluorescent timers which are fluorescent protein fragments that change their color as the protein populations age. Based on foregone information of the maturation kinetics of the fluorescent protein probes, the protein degradation rate, half-life, and other pharmacokinetic constants can be obtained by fitting the data to computational models.
- Tandem fluorescent protein fusion
Tandem fluorescent protein fusion is a new-developed strategy to assay protein half-life in vitro based on a conventional fluorescent protein timer. A novel feature of this method is that each moiety exhibits a distinct maturation time, which has circumvented the major limitations of the original fluorescent timers. Under steady-state conditions, the average age of the proteins directly reflects the protein half-life, irrespective of the synthesis rates.
Due to low-autofluorescence, non-toxicity, and convenient multiplexing, half-life assay by fluorescent has gradually become the most satisfying choice to research protein pharmacokinetics. If you are interested in our services, please feel free to contact us.
Reference
- Eldeeb, E. A.;et al. A molecular toolbox for studying protein degradation in mammalian cells. Journal of neurochemistry.2019, 151(4), 520-533.
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For Research Use Only. Not for Clinical Use.