Antibodies adsorption

To assess any antibody adsorption to Leica manufactured vials, a primary antibody was stored in the reagent vials for up to 1 mo at 4°C. Subsequently, the antibody was removed, and the remaining reagents of an immunostaining run were sequentially added, as in the Cadenza vial and Code-On isolon experiments. Slight vial staining was seen after the 1-mo incubation period, indicating minimal antibody adsorption. 

To assess possible adsorption of antisera to the Ventana dispensers, a Ventana pipeter filled with L26 antisera was stored at 4°C for up to 1 mo. At the end of the storage period, the antisera was decanted, and the pipeter thoroughly rinsed with PBS. The pipeter was then stained to assess antibody adsorption using sequential application of standard immunostaining reagents. After 4 wk of storage, there was no detectable immunoreactivity in the Ventana pipeter. 

TABLE 7 Ligand Selection for Antibody Adsorption by Affinity Related Mechanisms... 

Step 1 Capture antibody adsorption to solid-phase support... 

In heterogeneous assays, the detection is usually performed in a competitive format, in which labelled antigen is added to the sample solution, or in a sandwich format, in which after washing the siuface, secondary labelled antibodies are specifically adsorbed onto the immobilized antigens. For the latter format, the antigen must have two different epitopes—one for adsorption onto the capture antibody and one for the secondary antibody adsorption ... 

Acquired von Willebrand disease is a rare bleeding disorder that is similar to the congenital form of the disease. It has been reported primarily in association with autoimmune disorders, such as systemic lupus erythematosus, lymphoproliferative disorders, and neoplastic disease. Certain medications also have been associated with acquired von Willebrand disease, including valproic acid, dextran, and ciprofloxacin. Bleeding manifestations vary frommild to severe, and the condition often resolves with treatment of the underlying disease. In the acquired disease, von Wfllebrand factor appears to be synthesized in normal amounts, but then is rapidly removed from plasma by anti-von Willebrand factor antibodies, adsorption to tumor cells, or other mechanisms. ... 

In several series of binary mixtures, relative concentration of each protein in solution determined which was adsorbed most (15), but relationships were not simply predictable. For example, judged by optical thickness of deposited antibody, adsorption of fibrinogen in presence of IgG rapidly rose to its maximum as the relative concentration of fibrinogen was increased, while in presence of albumin fibrinogen adsorption increased almost linearly as the proportion of fibrinogen to albumin in solution was increased (15). 

Figure 3.8 The most frequently used methods to immobilize capture molecules onto microarray surfaces. (A) Antibody adsorption by poly-L-lysine-, nitrocellulose-, or polyvinylidene fluoride-treated surface. (B) Covalent binding using various silane reagents of APTES (3-aminopropyl)triethoxysilane, GPTS (3-gly-cidoxypropyl)trimethoxysilane, and MPTS (3-mercaptopropyl)trimethoxysilane. (C) Affinity interactions by biotin/streptavidine or histidine-tag/nickel-nitrilotri-acetic acid. (D) Diffusion-based (hydrogel) antibody-immobilization technique.

ZhouJ, Tsao HK, Sheng YJ, et al Monte Carlo simulations of antibody adsorption and orientation on charged surfaces, J Chem Phys 121 1050-1057, 2004a. 

A decrease of 0/C and P/C concentration ratios at the surface of Streptococcus sobrims was reported as a result of preincubation with polyclonal antibodies. A combination of the pH-dependent zeta potential and the XPS data suggested that polyclonal antibody adsorption occurred through blocking of surface phosphate. XPS measurements provided evidence for IgG attachment to Pseudomonas aeruginosa. The antibody-modified cells showed a reduced adhesion to glass under flow in a nutrient medium. ... 

Imbert-Laurenceau, E., Berger, M.C., Pavon-Djavid, G., Jouan, A., and Migonney, V. 2005. Surface modification of polystyrene particles for specific antibody adsorption. Polymer 46 1277-85. 

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