Preparatıon of Cryogel Columns for Depletıon of Hemoglobın from Human Blood

Proteins are vital molecules for the living organisms. They carry out many important physiological metabolic pathways. Therefore they have always been the study of interest. The term proteome defines the large-scale study of proteins including the proteins produced by any organism as well as their modifications made after the synthesis. An evolving field of the proteomics is to identify disease linked novel proteins within the erythrocyte proteins. As in the all proteomic studies of the blood plasma, high abundant proteins in erythrocytes, i.e. hemoglobin and carbonic anhydrase, interfere with the analysis of low abundant and disease related proteins. Thus it is essential to remove these proteins before detailed analysis of cytosolic proteins of erythrocytes. In the previous studies nickel affinity chromatography has been investigated for its efficiency for hemoglobin depletion and shown to be effective for increasing the number of detectable proteins.Proteinler canlı organizmalar için yaşamsal öneme sahip moleküllerdir. Çok sayıda önemli fizyolojik metabolik yolakları yürütürler. Dolayısıyla her zaman yoğun bir ilgiyle çalışılmaktadırlar. Proteom terimi herhangi bir organizmanın ürettiği proteinlerin, sentezlendikten sonraki modifikasyonlar da dahil, çalışılmasını ifade etmektedir. Proteomik çalışmalarının gelişen alanlarından bir tanesi de eritrosit proteinleri içerisinde hastalıklarla ilişkili yeni olanların tanımlanmasıdır. Kan plazmasının proteomik analizinde olduğu gibi, eritrositlerdeki fazla miktarda bulunan proteinler, hemoglobin ve karbonik anhidraz gibi, az miktarda bulunan ve hastalıklarla ilişkili proteinlerin görülmesini engellemektedir. Daha önceki çalışmalarda nikel afinite kromatografisi hemoglobin uzaklaştırma etkinliği bakımından incelenmiş ve tayin edilebilir protein sayısının artması için etkili olduğu gösterilmiştir

Molecularly imprinted hydrophobic polymers as a tool for separation in capillary electrochromatography

The use of molecular imprinted polymers (MIPs), which provides a means for preparing stationary phases with predetermined selectivity for a target molecule in capillary electrochromatography (CEC), is attractive because it combines selectivity, higher separation efficiency and shorter analysis time. A bisphenol A (BPA)-imprinted monolithic capillary BPA/PMAPA column was synthesized for the selective separation of bisphenol A (BPA) from aqueous solutions containing the competitor molecule phenol (PH), which is similar in size and shape to the template molecule. BPA-imprinted monolithic column was prepared in the presence of the template molecule, BPA, which results in the formation of recognition cavities complementary to the template molecule, after the removal of template molecule. An amino acid based monomer, N-methacryloyl-L-phenyl alanine (MAPA), was used as the functional monomer. The new stationary phase contains both charged and hydrophobic groups originating from MAPA monomer, which behaves as both an electroosmotic flow (EOF) supplier and a hydrophobic matrix. The MAPA containing BPA imprinted column behaves as a mixed mode stationary phase, as ion exchanger and hydrophobic matrix depending on the pH of the medium. Scanning electron microscopy was used to identify the structural features of the molecular imprinted column. MIP column performance was evaluated by the electrochromatographic separation of alkylbenzenes. The novelty of this work originated from dual separation mechanism shown by MAPA, which has the ability to form both hydrophobic and electrostatic interactions by the charged and hydrophobic groups of phenylalanine amino acid. This new column with mixed-mode characteristics was then used successfully as the stationary phase in CEC for the selective separation of BPA in MIP systems

Triazine herbicide imprinted monolithic column for capillary electrochromatography

Trietazine was selectively separated from aqueous solution containing the competitor molecule cyanazine, which is similar in size and shape to the template molecule. Structural features of the molecularly imprinted column were figured out by SEM. The influence of the mobile-phase composition, applied electrical field, and pH of the mobile phase on the recognition of trietazine by the imprinted monolithic polymer has been evaluated, and the imprint effect in the trietazine-imprinted monolithic polymer was demonstrated by an imprinting factor. The optimized monolithic column resulted in separation of trietazine from a structurally related competitor molecule, cyanazine. In addition, fast separation was obtained within 6 min by applying higher electrical field, with the electrophoretic mobility of 2.97 x 10(-8) m(2)V(-1)s(-1) at pH 11.0