Assessment of Adsorption and Adhesion of Proteins to Polystyrene Microwells by Sequential Enzyme-Linked-Immunosorbent Assay Analysis

Presentation of Peptide Antigens as Albumin Conjugates for Use in Detection of Serum Antibodies by Enzyme-Linked Immunosorbent Assay

“…The maximal level of protein adsorption, which was reached when 2.5 µg per 100 µL was added to a well, was about 70 ng/well, as determined by bicinchoninic acid protein assay (11). This value is close to previous findings (12). Similar to the dependence of ELISA response on the quantity of conjugate adsorbed to the well, an increase in response with the relative content of antigenic peptide in the conjugate was observed (Figure 1).…”

Development of a Competition Assay for the Evaluation of the Binding of Human Parotid Salivary Proteins to Dietary Complex Phenols and Tannins Using a Peroxidase-Labeled Tannin

“…Figure 2 shows the relative binding of HRP-EGC, of HRP conjugated with linker molecule (1,4-butanediol diglycidyl ether), and of HRP alone, in the absence of any test competing tannin, both to plates coated with parotid salivary protein and to uncoated blank plates. The blank values, that is, nonspecific binding of HRP conjugates to the surface of the polystyrene plate in the absence of salivary proteins, were reduced to a low level by the addition of Tween 20 to wash and competition buffers to block such binding sites, a protocol that is commonly used in ELISA assays to overcome problems of nonspecific antibody binding (Wilkins Stephens et al, 1995). The coated plates treated with HRP-EGC gave a 50-fold higher level of HRP binding compared with those treated with the HRP linker, and no binding was detected with HRP alone, thus demonstrating that binding of HRP-EGC was strongly dependent on the conjugation with the tannin.…”

PM-IRRAS Investigation of the Interaction of Serum Albumin and Fibrinogen with a Biomedical-Grade Stainless Steel 316LVM Surface

“…14,15 Since a comprehensive understanding of the mechanisms of protein adsorption is essential for the continuous development of more biocompatible medical devices, the interfacial behavior of proteins has long attracted the interest of researchers. A wide variety of experimental techniques have been applied to study different aspects of protein interaction with solid surfaces, includingenzyme-linkedimmunosorbentassay, 16,17 radiolabeling, [18][19][20] ellipsometry, [21][22][23][24] total internal reflection fluorescence, 25,26 optical waveguide lightmode spectroscopy, 22,27,28 surface plasmon resonance, [29][30][31][32] neutron reflectivity, 33,34 electrochemical quartz crystal nanobalance, 35,36 quartz crystal microbalance, 22,37,38 electrochemical impedance spectroscopy (EIS), 39 X-ray photoelectron spectroscopy, 40,41 time-of-flight static secondary ion mass spectroscopy, 42 and IR-based techniques such as attenuated total reflection Fourier transform IR (FTIR), 2,18,43 grazing angle FTIR 5,18 and circular dichroism. 44 The purpose of this work is to investigate the interaction of two major plasma proteins, bovine serum albumin (BSA) and fibrinogen, with a biomedical-grade 316LVM stainless steel surface by means of the polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) technique.…”