title: Ultra-fast mass fingerprinting by high-affinity capture of peptides and proteins on derivatized poly(glycidyl methacrylate/divinylbenzene) for the analysis of serum and cell lysates.
abstract: The development of support materials in mass fingerprinting is an important task required for diagnostic markers in conjunction with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The material-based approach, which we introduced as material-enhanced laser desorption/ionization (MELDI), focuses not only on different functionalities, but also emphasizes the morphology, i.e. porosity or particle size of the carrier material. As a result, it provides a quick and sensitive platform for effective binding of peptides and proteins out of different biofluids, e.g. serum, spinal fluid, urine or cell lysates, and to subsequently analyze them with MALDI-TOF MS. This approach includes a built-in desalting step for serum protein profiling and is sensitive enough to detect proteins and peptides down to 100 fmol/microL. Here we co-polymerized glycidyl methacrylate (GMA) with divinylbenzene (DVB) using thermal polymerization to yield a GMA/DVB polymer for further modifications. Different affinities have been created, such as immobilized metal ion affinity (IDA-Cu2+), reversed-phase (RP) and anion-exchanger (AX) chromatography. The diverse derivatizations and the dispersity of the particles created by different chemical synthetic approaches were confirmed by characteristic infrared (IR) peaks. The polymerization carried out by non-stirring yielded an average pore radius of 6.1 microm (macro-pores) that enhanced the binding capacity enormously by offering enlarged surface areas. Moreover, atomic absorption spectrometry (AAS) provided the metal content loaded on iminodiacetic acid (IDA) in the case of poly(GMA/DVB)-IDA-Cu2+. To summarize, the optimized MELDI approach is sensitive in its performance, extremely fast and can be adapted to robotic systems for routine analysis, allowing sample preparation in less than 5 min in contrast to the conventional surface-enhanced laser desorption/ionization (SELDI) methods.
authors.affiliation: Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innsbruck, Austria.
authors.forename: Matthias
authors.initials: M
authors.lastname: Rainer
authors.forename: Najam-ul-Haq
authors.initials: NU
authors.lastname: Muhammad
authors.forename: Christian W
authors.initials: CW
authors.lastname: Huck
authors.forename: Isabel
authors.initials: I
authors.lastname: Feuerstein
authors.forename: Rania
authors.initials: R
authors.lastname: Bakry
authors.forename: Lukas A
authors.initials: LA
authors.lastname: Huber
authors.forename: Douglas T
authors.initials: DT
authors.lastname: Gjerde
authors.forename: Xiajuan
authors.initials: X
authors.lastname: Zou
authors.forename: Honggang
authors.initials: H
authors.lastname: Qian
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Du
authors.forename: Fang
authors.initials: F
authors.lastname: Wei-Gang
authors.forename: Yang
authors.initials: Y
authors.lastname: Ke
authors.forename: Günther K
authors.initials: GK
authors.lastname: Bonn
mesh_terms: D001798:Blood Proteins / Q000737:chemistry*; D006528:Carcinoma, Hepatocellular / Q000737:chemistry / Q000473:pathology; D002457:Cell Extracts / Q000737:chemistry*; D045744:Cell Line, Tumor; D006801:Humans; D008113:Liver Neoplasms / Q000737:chemistry / Q000473:pathology; D008875:Middle Aged; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000097:blood*; D011109:Polymethacrylic Acids / Q000737:chemistry*; D011145:Polyvinyls / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D001798:Blood Proteins; D002457:Cell Extracts; D010455:Peptides; D011109:Polymethacrylic Acids; D011145:Polyvinyls; C516630:poly(glycidyl methacrylate-co-divinylbenzene)