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Synthesis and Identification of FITC-Insulin Conjugates Produced Using Human Insulin and Insulin Analogues for Biomedical Applications

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Abstract

Human insulin was fluorescently labelled with fluorescein isothiocyanate (FITC) and the conjugate species produced were identified using high performance liquid chromatography and electrospray mass spectroscopy. Mono-labelled FITC-insulin conjugate (A1 or B1) was successfully produced using human insulin at short reaction times (up to 5 h) however the product always contained some unlabelled native human insulin. As the reaction time was increased over 45 h, no unlabelled native human insulin was present and more di-labelled FITC-insulin conjugate (A1B1) was produced than mono-labelled conjugate with the appearance of tri-labelled conjugate (A1B1B29) after 20 h reaction time. The quantities switch from mono-labelled to di-labelled FITC-insulin conjugate between reaction times 9 and 20 h. In the presence of phenol or m-cresol, there appears to be a 10 % decrease in the amount of mono-labelled conjugate and an increase in di-labelled conjugate produced at lower reaction times. Clinically used insulin analogues present in commercially available preparations were successfully fluorescently labelled for future biomedical applications.

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Acknowledgments

We thank EPSRC National Mass Spectrometry Facility, Swansea University for their help with FITC-insulin analysis. D Jacob would like to acknowledge the research studentship provided by De Montfort University.

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Authors and Affiliations

  1. Leicester School of Pharmacy, Faculty of Health and Life Sciences, De Montfort University, The Gateway, Leicester, LE1 9BH, UK

    Dolly Jacob, M. Joan Taylor, Paul Tomlins & Tarsem S. Sahota

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  1. Dolly Jacob
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  2. M. Joan Taylor
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  3. Paul Tomlins
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  4. Tarsem S. Sahota
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Correspondence to Tarsem S. Sahota.

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Jacob, D., Joan Taylor, M., Tomlins, P. et al. Synthesis and Identification of FITC-Insulin Conjugates Produced Using Human Insulin and Insulin Analogues for Biomedical Applications. J Fluoresc 26, 617–629 (2016). https://doi.org/10.1007/s10895-015-1748-1

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  • DOI: https://doi.org/10.1007/s10895-015-1748-1

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