| 1. |
- Jagadeesan, Kishore, et al.
(författare)
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Filter Plate–Based Screening of MIP SPE Materials for Capture of the Biomarker Pro-Gastrin-Releasing Peptide
- 2017
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Ingår i: SLAS Discovery. - : Sage Publications. - 2472-5560 .- 2472-5552. ; 22:10, s. 1253-1261
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Tidskriftsartikel (refereegranskat)abstract
- Affinity-based solid-phase extraction (SPE) is an attractive low-cost sample preparation strategy for biomarker analysis. Molecularly imprinted polymers (MIPs) as affinity sorbents offer unique opportunities for affinity SPE, due to their low manufacturing cost and high robustness. A limitation is the prediction of their affinity; therefore, screening of analyte recovery and specificity within a large range of SPE conditions is important in order to ensure high-sensitivity detection and assay reproducibility. Here, a µ-SPE method for screening of the MIP-SPE materials using a commercial 384-well filter plate is presented. The method allows for rapid and automated screening using 10?30 µL of packed SPE sorbent per well and sample volumes in the range of 10?70 µL. This enables screening of many different SPE sorbents while simultaneously identifying optimal SPE conditions. In addition, the 384-well format also facilitates detection with a multitude of analytical platforms. Performance of the µ-MIP-SP
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| 2. |
- Qader, Abed Abdel, et al.
(författare)
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Peptide imprinted receptors for the determination of the small cell lung cancer associated biomarker progastrin releasing peptide
- 2014
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Ingår i: Journal of Chromatography A. - : Elsevier. - 0021-9673 .- 1873-3778. ; 1370, s. 56-62
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Tidskriftsartikel (refereegranskat)abstract
- Peptide imprinted polymers were developed for detection of progastrin releasing peptide (ProGRP); a low abundant blood based biomarker for small cell lung cancer. The polymers targeted the proteotypic nona-peptide sequence NLLGLIEAK and were used for selective enrichment of the proteotypic peptide prior to LCMS based quantification. Peptide imprinted polymers with the best affinity characteristics were first identified from a 96-polymer combinatorial library. The effects of functional monomers, crosslinker, porogen, and template on adsorption capacity and selectivity for NLLGLIEAK were investigated and optimized. Ultimately, a solid phase extraction method was developed for highly selective enrichment of the target peptide from tryptic digests.
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| 3. |
- Rossetti, Cecilia, et al.
(författare)
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Antibody-free biomarker determination : exploring molecularly imprinted polymers for pro-gastrin releasing Peptide
- 2014
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 86:24, s. 12291-12298
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Tidskriftsartikel (refereegranskat)abstract
- Biomarker mass spectrometry-assays are highly demanded and analysis of Pro-Gastrin Releasing Peptide (ProGRP) as Small Cell Lung Cancer marker has been recently investigated by mass spectrometry after immuno-extraction. In this paper we introduce an assay based on molecularly imprinted polymers (MIPs) targeting the proteotypic peptide of ProGRP, as a possible alternative to current immuno-based assay. The MIPs were prepared by surface initiated Reversible Addition-Fragmentation chain Transfer (RAFT) polymerization and were introduced as sorbents for clean-up and enrichment of ProGRP signature peptide from tryptically treated serum samples. The use of an appropriate solid phase extraction (SPE) protocol allowed specific extraction of the target peptide while depleting other peptides risen from the sample digestion, hence resulting in a reduced background. The selective extraction of ProGRP signature peptide, after serum samples digestion, translates into a time and cost-effective method suited for bottom-up analysis wherever targeted peptide extraction from complex matrices is required
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| 4. |
- Rossetti, Cecilia, et al.
(författare)
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Automated Protein Biomarker Analysis : on-line extraction of clinical samples by Molecularly Imprinted Polymers
- 2017
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Ingår i: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 7
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Tidskriftsartikel (refereegranskat)abstract
- Robust biomarker quantification is essential for the accurate diagnosis of diseases and is of great value in cancer management. In this paper, an innovative diagnostic platform is presented which provides automated molecularly imprinted solid-phase extraction (MISPE) followed by liquid chromatography-mass spectrometry (LC-MS) for biomarker determination using ProGastrin Releasing Peptide (ProGRP), a highly sensitive biomarker for Small Cell Lung Cancer, as a model. Molecularly imprinted polymer microspheres were synthesized by precipitation polymerization and analytical optimization of the most promising material led to the development of an automated quantification method for ProGRP. The method enabled analysis of patient serum samples with elevated ProGRP levels. Particularly low sample volumes were permitted using the automated extraction within a method which was time-efficient, thereby demonstrating the potential of such a strategy in a clinical setting.
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| 5. |
- Rossetti, Cecilia, et al.
(författare)
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Exploring the peptide retention mechanism in molecularly imprinted polymers
- 2017
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Ingår i: Analytical and Bioanalytical Chemistry. - : Springer. - 1618-2642 .- 1618-2650. ; 409:24, s. 5631-5643
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Tidskriftsartikel (refereegranskat)abstract
- Molecularly imprinted polymers (MIPs) have been used as useful sorbents in solid-phase extraction for a wide range of molecules and sample matrices. Their unique selectivity can be fine-tuned in the imprinting process and is crucial for the extraction of macromolecules from complex matrices such as serum. A relevant example of this is the application of MIPs to peptides in diagnostic assays. In this article the selectivity of MIPs, previously implemented in a quantitative mass-spectrometric assay for the biomarker pro-gastrin-releasing peptide, is investigated. Partial least squares regression was used to generate models for the evaluation and prediction of the retention mechanism of MIPs. A hypothesis on interactions of MIPs with the target peptide was verified by ad hoc experiments considering the relevant peptide physicochemical properties highlighted from the multivariate analysis. Novel insights into and knowledge of the driving forces responsible for the MIP selectivity have been obtained and can be directly used for further optimization of MIP imprinting strategies. Graphical Abstract Applied analytical strategy: the Solid Phase Extraction (SPE) of digested Bovin Serum Albumin (BSA), using Molecularly Imprinted Polymers (MIP), is followed by the liquid chromatography-mass spectrometry (LC-MS) analysis for the identification of the retained peptides. The further application of multivariate analysis allows setting up a Partial Least Square (PLS) model, which describes the peptide retention into the MIP and gives additional knowledge to be used in the optimization of the MIP and the whole SPE method.
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