Proteomic analysis of protein adsorption: Serum amyloid P adsorbs to materials and promotes leukocyte adhesion

Abstract

Serum and plasma protein adsorption on materials was analyzed using gel electrophoresis and ion trap mass spectrometry. Following incubation of polypropylene, polyethylene terephthalate (PET), or polydimethylsiloxane (PDMS) with 5% serum for longer than 4 h, we found unexpectedly high amounts of the pentraxin serum amyloid P. It was previously shown that serum amyloid P is constitutively expressed in humans, functions as an opsonin, and interacts with the Fcgamma receptors on leukocytes. We demonstrate that serum amyloid P adsorbed to tissue culture polystyrene, PDMS, and PET promotes the adhesion of granulocytes and monocytes in the presence of calcium. The methods developed for these studies may be useful for the large-scale study of protein adsorption and do not rely on radiolabeling or the availability of antibodies.

Binding of human serum amyloid P componentto L‐selectin

“…Immobilization of either SAP [30,31] or Lselectin [32] may enhance their affinity for respective ligands. SAP has recently been described to be highly absorbed from plasma on biomedical materials such as polystyrene, promoting Ca 2+ -dependent leukocyte adhesion [31].…”

“…Immobilization of either SAP [30,31] or Lselectin [32] may enhance their affinity for respective ligands. SAP has recently been described to be highly absorbed from plasma on biomedical materials such as polystyrene, promoting Ca 2+ -dependent leukocyte adhesion [31]. We found previously that circulating Lselectin plasma concentrations drop by almost 50% in infants undergoing open heart surgery employing cardiopulmonary bypass [33], which may be due to trapping of soluble L-selectin by SAP absorbed onto the surface of plastic tubings.…”

Quantitation of nonspecific protein adsorption at solid–liquid interfaces for single-cell proteomics

“…Besides using MS for direct analysis at the surface, by stripping the proteins off the surface and using shotgun proteomics 133 (detection and quantification of unknown proteins based on sequencing their derived peptides), hundreds and thousands of types of proteins can be quantified simultaneously, which provided unprecedented information for the biofouling on medical implants [134][135][136][137] . Examples of these applications include the 2D-D r a f t gel MS as mentioned in the SDS-PAGE section and gel-free liquid chromatography separation and MS analyses.…”

Mass spectrometric mapping of fibrinogen conformations at poly(ethylene terephthalate) interfaces☆

“…Each experiment was immediately followed by a blank injection to ensure that proteins were not carried over from previous injections. Data were collected in full MS/MS mode and biotin-labeled peptides were identified using an in-house program as described previously [40].…”

“…We have developed a proteomics-based strategy to analyze the adsorption of bFg onto PET, which supplements our earlier proteomic studies on the adsorption of serum and plasma proteins on biomaterials [40]. The conformational changes that occur upon fibrinogen adsorption were examined by comparing the mass spectra of bFg that was biotin-labeled in solution to bFg that was labeled after adsorption to the surfaces of PET particles.…”