Anti-protein antibodies

PETTY, R.E. STEWARD, M.W. (1977) The effect of immunological adjuvants on the relative affinity of anti-protein antibodies. Immunology, 32,49-55. 

Incidences The following short list includes murine, chimeric, and humanized monoclonal antibodies as well as interferons and interleukins against which anti-protein antibodies have been raised in patients ... 

The route of application has a substantial impact on eliciting anti-protein antibodies. Intramuscular and subcutaneous applications seem to be more prone to raise anti-protein antibodies as compared to intravenous application [58, 59]. 

In summary, it is of utmost interest to determine the level of anti-protein antibodies and their impact on the in vivo situation in order to allow the implementation of countermeasures to minimize immune reactions. One major... 

Alternatively to direct measurement of anti-protein antibodies, there seems to be an increased interest in the use of T-cell responses as a marker of immunogenic-ity [64] (see also below). 

Peptide-MHC Interaction Analysis The development of an anti-protein antibody response frequently requires activation of T-helper cells that induce B-cells to secrete specific antibodies. The key unit for regulation of this process is a trimolecular complex consisting of the T-cell receptor on T-cells, and the class II major histocompatibility complex (MHC) harboring an MHC-ligand on antigen presenting cells. The MHC-ligands are peptides derived... 

ELP fusion protein (Fig. 9, center). It was shown that picomolar levels of ELP fusion proteins could be purified via ELP coaggregation [47, 48]. The value of this technique was shown by the purification of low levels of ELP fused to an anti-atrazine antibody [49]. 

B) Proteins were resolved by SDS-PAGE, blotted and PG2 polypeptides detected by reaction with anti-PG2 antibodies. Lane 1, 2 pg purified PGl Lane 2, 1 pg purified PG2 Lane 3, 1 pg purified Psubunit. 

Studies with polyclonal antibodies against the 0.2 d protein have identified neuronal 0.2 polypeptides [58] and other studies with anti-similar protein in brain [72]. Future studies will be aimed at further clarifying the similarities and differences in the structures of L channel proteins from various sources. 

We used an anti-DNA antibody as an exploratory model system. The antibody was monoelonal from mouse sourees and its subelass was IgM. Mouse IgG (MW 1.5 x 10 Da) and IgM (MW 9 X 10 Da) antibodies from normal plasma, and bovine serum albumin were used for the eontrol measurements. To prevent the nonspeeilie adsorption of proteins to the uneovered, bare Au site in the modified eleetrode surfaee, the DNA-modified eleetrode prepared by the standard proeedure was further treated with aqueous 2-mercaptoethanol solution and was used for the measurements. 

CV measurements showed that the reversible eleetrode reaetion of the [Fe(CN)6]" redox eouple was suppressed to some extent by the treatment with the DNA. The addition of the anti-DNA antibody further suppressed the redox reaetion thus decreasing the magnitudes of the CV peak currents. This is most likely caused by a steric hindrance of the bulky protein, which binds to the DNA double strands on the electrode surface, to mainly reduce the effective area of the electrode. The electrostatic repulsive effect may also contribute to the electrode response, since the isoelectric point of mouse IgM is commonly in the range of 4.5 to 7.0. Figure 11 shows the relationship between the decrease in the anodic peak current (A/p ) and the antibody concentration. As seen in this figure, the electrode system responded to the anti-DNA antibody in the concentration range of 1 — 100 nM. For the case of the mouse IgM, which does not interact with double-stranded DNA, the present system gave almost no response. The sensor did not respond to other serum proteins as well (data not shown). 

The endothelin B receptor is an example of characterization of a homogeneous, affinity purified protein (Roos et al., 1998). Significant progress has been made in the development of techniques for more high-throughput identification of phosphorlyation events. Analysis of large sets of phosphorylated proteins is facilitated by the availability of affinity purification methods such as anti-phosphotyrosine or anti-phosphoserine antibodies or metal affinity chromatography (Neubauer and Mann, 1999 Soskic et al., 1999). These methods are not specific to a particular protein but rather are used to fractionate all proteins that are phosphorylated. 

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