title: Advance workflow in lipoproteomics via polymeric ion exchanger.
abstract: A workflow is designed for the analysis of lipoproteins, high density lipoproteins (HDL), apoproteins, and lipid fraction, employing an organic polymeric anion exchanger through the enrichment of lipoproteins/peptides from serum. Polymeric separation media are chemically stable over the wide pH range. Poly(GMA/DVB), poly(GMA/EGDMA), and poly(GPE/DVB) are synthesized by radical polymerization, derivatized as strong anion exchangers, and used for lipoproteins enrichment. Lipoprotein's surface is covered by phospholipids, having phosphate groups, therefore lipoproteins are enriched by the interaction of anion exchanger with the phosphate groups and eluted at the pH of 7.5. HDL are further isolated by precipitating the very low-density lipoproteins (VLDL) and low-density lipoproteins (LDL) with phosphotungstic acid as precipitating reagent, followed by delipidation via liquid/liquid extraction. Apolipoproteins profiling is done by MALDI-MS, and lipids are analyzed using gold nanoparticles in the LDI-MS process. This study introduces a lipoproteomics work flow in separation science which analyses the intact lipoproteins. Furthermore, solid phase extraction (SPE)-based methodology is reported for the first time in lipoproteomics. Use of organic polymers, high reproducibility, detailed analysis of lipoproteins, apoproteins/peptides, and lipids from the single serum sample are the distinctive features of this workflow. Being biomarkers of numerous diseases, lipoproteins have clinical significance, and this workflow can be used at diagnostic and therapeutic levels.
authors: Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800, Pakistan.
authors: Rabia
authors: R
authors: Javeed
authors: Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800, Pakistan.
authors: Fahmida
authors: F
authors: Jabeen
authors: Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800, Pakistan.
authors: Hira
authors: H
authors: Saeed
authors: Division of Analytical Chemistry, Institute of Chemical Sciences, Bahauddin Zakariya University, Multan 60800, Pakistan.
authors: Muhammad
authors: M
authors: Najam-ul-Haq
mesh_terms: D006801:Humans; D006863:Hydrogen-Ion Concentration; D007474:Ion Exchange; D008074:Lipoproteins / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013499:Surface Properties
chemical_list: D008074:Lipoproteins; D011108:Polymers