Facile preparation of organic-inorganic hybrid polymeric ionic liquid monolithic column with a one-pot process for protein separation in capillary electrochromatography.

An organic-inorganic hybrid monolithic column based on 1-vinyl-3-dodecylimidazolium bromide (VC12Im(+)Br(-)) has been prepared in a single step by combining radical copolymerization with a non-hydrolytic sol-gel (NHSG) process. The NHSG process was significantly shortened to 6 h by using formic acid as catalyst. For comparison, we also prepared polymeric ionic liquid (PIL) monolithic columns by hydrolytic sol-gel and organic polymeric process, respectively. The resulting monolithic columns were characterized by Fourier transform infrared spectra, scanning electron microscopy, and Brunauer-Emmett-Teller. Under the capillary electrochromatography mode, these columns were applied to separate alkylbenzenes, anilines, and proteins, respectively. The results indicated that the NHSG-based hybrid PIL monolithic column exhibited the highest column efficiency among the three types of columns; organic solvent, commonly required by the traditional columns to achieve satisfactory separation efficiency for proteins, was absent in the NHSG-based hybrid PIL monolithic column because of the biocompatibility of the VC12Im(+)Br(-), which was beneficial to analysis of protein containing samples. In order to demonstrate its application potential, the developed NHSG-based hybrid PIL monolithic column was also employed to separate egg white sample.

Fluorescence sensor based on molecularly imprinted polymers and core-shell upconversion nanoparticles@metal-organic frameworks for detection of bovine serum albumin.

A novel strategy for sensing protein was proposed through combining the high selectivity of molecular imprinting technology with the excellent upconversion fluorescence of upconversion nanoparticles (UCNPs) and high specific surface area of metal-organic frameworks (MOFs). Herein, the UCNPs acted as signal reporter and MOFs were introduced to increase the rate of mass transfer. The UCNPs@MIL-100 as support material was prepared via a step-by-step method. The imprinted material-coated UCNPs@MIL-100 (UCNPs@MIL-100@MIPs) were obtained by sol-gel technique. The results showed that as the increase of the template protein concentration, the fluorescence intensity of UCNPs@MIL-100@MIPs quenched gradually, and the imprinting factor was 2.90. The linear in the range of 1.00 to 8.00 µM, and the detection limit was 0.59 µM. Therefore, the novel optosensing material is very promising for future applications.

Macroporous molecularly imprinted monolithic polymer columns for protein recognition by liquid chromatography.

Macroporous cytochrome c (cyc)-imprinted monolithic polyarylamide columns were prepared, and applied for the template protein recognition by HPLC. With cyc (18.8 mg) as template, the imprinted monolithic materials were in situ polymerized in an HPLC column tube, with methacrylamide (450 mg), methacrylic acid (15.8 mg), piperazine diacrylamide (720 mg) and ammonium sulfate (390 mg) dissolved in 5 mL of phosphate buffer (pH 7.4), initiated by ammonium persulfate and TEMED. After the reaction, cyc was removed with acetic acid (10%, v/v) containing 10% w/v SDS. The non-imprinted monolithic column was prepared under the same procedure except without cyc. Retention of cyc and its competitive protein, lysozyme (lys), on molecular-imprinting polymer (MIP) and non-imprinted polymer columns was studied. When the pH value of mobile phase was 4.0, on MIP column, the retention factors of cyc and lys were 2.0 and 1.3, respectively. However, those on non-imprinted polymer column were very similar, both as 1.1. Even in competitive environment, a mixture of cyc and lys could be separated on MIP column under gradient elution, with resolution as 1.2. These results indicate that protein-imprinted monolithic polymer columns could offer obvious affinity and specific recognition to the template protein.

One-step post-imprint modification achieve dual-function of glycoprotein fluorescent sensor by "Click Chemistry".

A novel molecular imprinting fluorescent sensor synthesized avoiding complicated design and synthesis of a fluorescent functional monomer, and without the extra cost of additional fluorescence materials (quantum dots, carbon dots and fluorescein), only by a click reaction making the common protein imprinting polymers upgraded to a fluorescent sensing material which can be applied to the fast detection of glycoproteins. In this study, a common protein imprinted polymer (protein-IP) which containing a disulfide bond in the imprinted cavities was first synthesized, then the disulfide linkage was cleaved by reduction, the exposed thiol groups were used for the click reaction to introduce 4-vinyl phenylboronic acid to improve the imprinted recognition for glycoproteins and convert recognition behavior into fluorescent signal simultaneously in one-step site-specific post-imprinting modifications (PIMs). Good linear relationship was obtained in the range of 0.1-10mgmL-1 by fluorescence assay, comparison of the adsorption capacity of the MIPs before and after modification proved that the method has no negative impact on the imprinting effect and exhibit good imprinting effect for glycoproteins of different molecular size and weight.

Upconversion fluorescence metal-organic frameworks thermo-sensitive imprinted polymer for enrichment and sensing protein.

A novel fluorescence material with thermo-sensitive for the enrichment and sensing of protein was successfully prepared by combining molecular imprinting technology with upconversion nanoparticles (UCNPs) and metal-organic frameworks (MOFs). Herein, the UCNPs acted as signal reporter for composite materials because of its excellent fluorescence property and chemical stability. MOFs were introduced to molecularly imprinted polymer (MIP) due to its high specific surface area which increases the rate of mass transfer relative to that of traditional bulk MIP. The thermo-sensitive imprinted material which allows for swelling and shrinking with response to temperature changes was prepared by choosing Bovine hemoglobin (BHB) as the template, N-isopropyl acrylamide (NIPAAM) as the temperature-sensitive functional monomer and N,N-methylenebisacrylamide (MBA) as the cross-linker. The recognition characterizations of imprinted material-coated UCNPs/MOFs (UCNPs/MOFs/MIP) were evaluated, and the results showed that the fluorescence intensity of UCNPs/MOFs/MIP reduced gradually with the increase of BHB concentration. The fluorescence material was response to the temperature. The adsorption capacity was as much as 167.6 mg/g at 28°C and 101.2mg/g at 44°C, which was higher than that of traditional MIP. Therefore, this new fluorescence material for enrichment and sensing protein is very promising for future applications.

Metal-organic frameworks supported surface-imprinted nanoparticles for the sensitive detection of metolcarb.

A novel approach to synthesize molecularly imprinted polymer (MIP) nanoparticles using a MIL-101 support (a type of metal-organic framework) is reported herein for the first time; the sample is referred as MIL@MIP. The nanoparticles were well distributed within the polymer film, and exhibit an octahedral shape, satisfied thermal stability, and a high specific surface area (SSA) of 1579.43 m(2)g(-1). The adsorption behavior of MIL@MIP toward metolcarb in aqueous solution was subsequently examined. The synthesized MIL@MIP displayed satisfactory high transfer mass rates and a high selective adsorption affinity for metolcarb. Based on these results, a quartz crystal microbalance (QCM) sensor based on MIL@MIP was subsequently constructed and examined for the sensitive detection of metolcarb. Under optimal conditions, the detection limit of the system assessed in pear juice was 0.0689 mg L(-1) within a linear concentration range of 0.1-0.9 mg L(-1). MIL@MIP-QCM system combines the advantages of MIL-101 and molecularly imprinted technology (MIT), thereby achieving high detection sensitivity and selectivity. The current findings suggest the potential of MIL@MIP for detecting trace level pesticides and veterinary drugs for food safety and environmental control.

A Novel Poly(ionic liquid) Interface-Free Two-Dimensional Monolithic Material for the Separation of Multiple Types of Glycoproteins.

Currently, many types of affinity materials have been developed for the enrichment of glycoproteins potentially considered to be clinical biomarkers; however, they can not effectively distinguish between different glycoproteins and thus lack the functionality that may be the key to the diagnosis of specific diseases. In the present work, a novel interface-free 2D monolithic material has been developed for the separation of multiple types of glycoproteins, in which boronate-functionalized graphene acts as preconcentration segment and poly(guanidinium ionic liquid) acts as separation segment. The resultant 2D material was characterized by X-ray photoelectron spectroscopy, elemental analysis, and electroosmotic flow analysis to demonstrate successful modification at each step. The performance of this 2D material was evaluated by capillary electrochromatography and allowed the successful online concentration and separation of five standard glycoproteins. The high separation efficiency can be largely attributed to the good orthogonality of boronate-functionalized graphene monolith and poly(guanidinium ionic liquid) monolith.

Preparation of protein imprinted materials by hierarchical imprinting techniques and application in selective depletion of albumin from human serum.

Hierarchical imprinting was developed to prepare the protein imprinted materials, as the artificial antibody, for the selective depletion of HSA from the human serum proteome. Porcine serum albumin (PSA) was employed as the dummy template for the fabrication of the recognition sites. To demonstrate the advantages of the hierarchical imprinting, molecularly imprinted polymers prepared by hierarchical imprinting technique (h-MIPs) were compared with those obtained by bulk imprinting (b-MIPs), in terms of the binding capacity, adsorption kinetics, selectivity and synthesis reproducibility. The binding capacity of h-MIPs could reach 12 mg g(-1). And saturation binding could be reached in less than 20 min for the h-MIPs. In the protein mixture, h-MIPs exhibit excellent selectivity for PSA, with imprinting factors as about 3.6, much higher than those for non-template proteins. For the proteomic application, the identified protein group number in serum treated by h-MIPs was increased to 422, which is 21% higher than that obtained from the original serum, meanwhile the identified protein group number for the Albumin Removal kit was only 376. The results demonstrate that protein imprinted polymers prepared by hierarchical imprinting technique, might become the artificial antibodies for the selective depletion of high abundance proteins in proteome study.

Highly sensitive fluorescent sensing for water based on poly(m-aminobenzoic acid).

Fluorescent conjugated polymer, poly(3-aminobenzoic acid), as a new class of fluorescence sensor for detection of trace amounts of water in organic solvents was developed. This sensor exhibits extraordinarily high sensitivity with a detection limit as low as 0.008 wt% for water in DMF.

(S)-Ibuprofen-imprinted polymers incorporating gamma-methacryloxypropyl-trimethoxysilane for CEC separation of ibuprofen enantiomers.

In this report, a novel preparation method of molecularly imprinted polymers (MIPs) for CEC was developed. Molecularly imprinted monolithic columns for (S)-ibuprofen were prepared and evaluated, in which charged entities responsible for establishing EOF have been derived from gamma-methacryloxypropyltrimethoxysilane (gamma-MAPS), which was hydrolyzed following copolymerization with 4-vinylpyridine (4-VPY) and ethylene glycol dimethacrylate (EDMA). The EOF and molecular recognition of the stationary phase were investigated in aqueous and nonaqueous media, respectively. The experimental results indicated that the material showed a reasonably stable EOF and the prepared separation materials were capable of separating racemic ibuprofen, a task that could not be accomplished by MIPs prepared in parallel, using methacrylic acid (MAA) as a functional monomer. The efficiency at pH 3.2 for the first-eluted enantiomer and the last-eluted enantiomer (the imprinted analyte) were 128,700 and 2100 plates/m, respectively.