Nonspecific binding elimination

Purify the iodinated protein from excess reactants by gel filtration using a desalting resin. The column may be pre-treated by passing a solution of bovine serum albumin (BSA) through it to eliminate nonspecific binding sites that could cause significant protein loss in small-sample applications. 

The use of silica particles in bioapplications began with the publication by Stober et al. in 1968 on the preparation of monodisperse nanoparticles and microparticles from a silica alkoxide monomer (e.g., tetraethyl orthosilicate or TEOS). Subsequently, in the 1970s, silane modification techniques provided silica surface treatments that eliminated the nonspecific binding potential of raw silica for biomolecules (Regnier and Noel, 1976). Derivatization of silica with hydrophilic, hydroxylic silane compounds thoroughly passivated the surface and made possible the use of both porous and nonporous silica particles in all areas of bioapplications (Schiel et al., 2006). 

The following protocol is based on the method of Morris and Saelinger (1984) for the labeling of succinylated avidin with gold particles of 5.2nm diameter. Succinylated avidin was used to reduce the pi of the protein, thus eliminating nonspecific binding due to the strong positive... 

With further work on sensor arraying, more effective means of sensor referencing may be developed to eliminate the effects not only of temperature but also nonspecific binding and test sample contamination. The overall objective is to create label-free optical sensor arrays that require minimal if any temperature stabilization, and as little a priori sample purification as possible. 

F(ab )2 fragments of antibodies are generated by pepsin digestion of whole IgG antibodies to remove a portion of the Fc region (see Sect. 1.1). Elimination of nonspecific binding of the Fc region of antibodies to Fc receptors on cell surfaces in... 

Since FPIAs are conducted as homogeneous immunoassays, they are susceptible to effects from endogenous fluorophores and from intersample variations. Such problems and others due to the sample matrix are largely avoided by sample dilutions of several hundredfold. Low-affinity, nonspecific binding of tracers to sample proteins, when present in sufficiently high concentrations, can result in a falsely elevated polarization signal. Interference from sample proteins can be eliminated when warranted, by proteolytic hydrolysis with pepsin.(46)... 

These results show that in a MISPE procedure one may encounter substances with different patterns of behavior. In the ideal case the template (or the template and a group of its analogs which need to be carried on for further analysis) are retained 100% during the sample application and the wash step. All other substances (or at least those which interfere with further analysis) should be fully removed. In the non-ideal case some interferents will not be removed by a particular protocol. In this case it is useful to observe the behavior of these interferents on the NIP. If they are retained by the NIP, too, then their retention is likely due to nonspecific binding. In this case a better choice of the solvents in the protocol may help to eliminate the interferent. The danger here is that the new protocol may also reduce the retention of the template and reduce its recovery. It is therefore interesting to note that even in this good paper it has not been shown that the addition of 1 % methanol to the dichloromethane sample was really necessary with the real sample matrices. 

Zheng M, Davidson F, Huang XY (2003) Ethylene glycol monolayer protected nanoparticles for eliminating nonspecific binding with biological molecules. J Am Chem Soc 125 7790-7791... 

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