Development of new multifunctional terpolymer sorbent for proteomics applications.
Determination of the availability of phases for specific separations is an important task achieved by a separation chemist. This becomes vital when the complex samples like biofluids are dealt with in proteome science. The work presented here involves the synthesis and application of terpolymeric sorbent with different functionalizations adopted for the selective enrichment of biomolecules of interest from biological fluids. Synthesis of terpolymer was carried out by the radical polymerization of monomers: methyl acrylate, acrylic acid and vinyl acetate with diethylene glycol dimethacrylate as cross-linking agent, benzoyl peroxide as initiator and chloroform as a porogenic solvent. Characterization was done through Fourier transform infrared spectroscopy, scanning electron microscopy and nitrogen adsorption porosimetry. The polymer was further modified to immobilized metal ion affinity chromatographic material, with immobilized Fe(3+)/La(3+) ions that allowed phosphopeptide enrichment from tryptic digests of standard proteins as well as milk, egg yolk and human serum. Sensitivity of enrichment down to 50 fmol was achieved in the presence of complex protein background as bovine serum albumin. Hydrophobicity was introduced through octadecyl amine, which provides comparable results to ZipTip C18/C4 for desalting of complex mixtures of all caseins. Analysis of the enriched content was performed by Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS).
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Muhammad
M
Najam-ul-Haq
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Adeela
A
Saeed
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Fahmida
F
Jabeen
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Dilshad
D
Hussain
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Naseem
N
Khan
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Maryam
M
Shabir
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Nadeem
N
Raza
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Muhammad Naeem
MN
Ashiq
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Muhammad Aslam
MA
Malana
Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
Zafar Iqbal
ZI
Zafar
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D004530:Egg Yolk / Q000737:chemistry; D008892:Milk / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
D010446:Peptide Fragments; D010750:Phosphoproteins; D011108:Polymers; D014357:Trypsin
IMAC (immobilized metal ion affinity chromatography); MALDI-MS; desalting; phosphoproteomics; polymer; radical polymerization; tryptic digests