FITC labeling of human insulin and transport of FITC-insulin conjugates through MDCK cell monolayer

Fluorescein isothiocyanate-labeled insulin (FITC-insulin) has been widely used for bioanalytical applications. Due to the high cost of commercial FITC-insulin and tedious labeling procedures described in the literature, there is still a need to develop a cost effective, reliable and quick labeling m...

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Main Authors: Darshana Shah (Author), Yuxing Guo (Author), Joseph Ocando (Author), Jun Shao (Author)
Format: Book
Published: Elsevier, 2019-12-01T00:00:00Z.
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042 |a dc 
100 1 0 |a Darshana Shah  |e author 
700 1 0 |a Yuxing Guo  |e author 
700 1 0 |a Joseph Ocando  |e author 
700 1 0 |a Jun Shao  |e author 
245 0 0 |a FITC labeling of human insulin and transport of FITC-insulin conjugates through MDCK cell monolayer 
260 |b Elsevier,   |c 2019-12-01T00:00:00Z. 
500 |a 2095-1779 
500 |a 10.1016/j.jpha.2019.08.002 
520 |a Fluorescein isothiocyanate-labeled insulin (FITC-insulin) has been widely used for bioanalytical applications. Due to the high cost of commercial FITC-insulin and tedious labeling procedures described in the literature, there is still a need to develop a cost effective, reliable and quick labeling method for insulin. The purpose of the present work was to develop a quick and affordable method for FITC labeling of human insulin and to determine the effect of different conjugations of FITC to human insulin on its permeability through the MDCK cell monolayer. FITC labeling of insulin gives mono-, di- or tri-conjugates depending on the reaction time and the molar ratio of FITC:insulin. Mono-conjugate with unlabeled insulin, mixture of di- and tri-conjugate, and tri-conjugate with very little amount of di-conjugate were synthesized in less than 4 h. Degree of conjugation had an effect on the permeability of insulin through the MDCK cell monolayer. Mono-conjugate had higher permeability than the unlabeled insulin due to increase in partition coefficient. However, tri-conjugate showed lower permeability than the unlabeled insulin due to the increase in molecular weight. Keywords: Insulin, FITC labeling, Conjugates, MDCK cells, Mono-conjugate, Tri-conjugate 
546 |a EN 
690 |a Therapeutics. Pharmacology 
690 |a RM1-950 
655 7 |a article  |2 local 
786 0 |n Journal of Pharmaceutical Analysis, Vol 9, Iss 6, Pp 400-405 (2019) 
787 0 |n http://www.sciencedirect.com/science/article/pii/S2095177919300668 
787 0 |n https://doaj.org/toc/2095-1779 
856 4 1 |u https://doaj.org/article/a56679a2d7e1412a895ee1ba3d7920a3  |z Connect to this object online.