Quantification and confocal imaging of protein specific molecularly imprinted polymers.
We have employed FITC--albumin as the protein template molecule in an aqueous phase molecular imprinted polymer (HydroMIP) strategy. For the first time, the use of a fluorescently labeled template is reported, with subsequent characterization of the smart material to show that the HydroMIP possesses a significant molecular memory in comparison to that of the nonimprinted control polymer (HydroNIP). The imaging of the FITC--albumin imprinted HydroMIP using confocal microscopy is described, with the in situ removal of the imprinted protein displayed in terms of observed changes in the fluorescence of the imprinted polymer, both before and after template elution (using a 10% SDS/10% AcOH (w/v) solution). We also report the imaging of a bovine hemoglobin (BHb) imprinted HydroMIP using two-photon confocal microscopy and describe the effects of template elution upon protein autofluorescence. The findings further contribute to the understanding of aqueous phase molecular imprinting protocols and document the use of fluorescence as a useful tool in template labeling/detection and novel imaging strategies.
Hawkins
Daniel M
DM
School of Biomedical and Molecular Sciences, University of Surrey, Guildford, Surrey, GU2 7XH, United Kingdom.
Trache
Andreea
A
Ellis
E Ann
EA
Stevenson
Derek
D
Holzenburg
Andreas
A
Meininger
Gerald A
GA
Reddy
Subrayal M
SM
D000418:Albumins / Q000737:chemistry*; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D001709:Biotechnology / Q000379:methods*; D002417:Cattle; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry*; D006454:Hemoglobins / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D017785:Photons; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013678:Technology, Pharmaceutical
D000418:Albumins; D001672:Biocompatible Materials; D006454:Hemoglobins; D011108:Polymers; D011506:Proteins; D016650:Fluorescein-5-isothiocyanate