Antibody concentrations serum

The concentration of antibody in tissue cultures of the hybridoma is low (10-60"gml- ) but the use of large culture vessels can obviate this. The hybridoma can also be propagated in mice where the antibody concentration in the serum and other body fluids can reach lOrngml. ... 

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). 

Antibody concentrations and affinities vary considerably. The optimal dilution for a given primary antibody must be determined empirically (although most companies will give an indicative range of dilutions). In general, early bleed serum or tissue culture supernatant are used at 1 100-1 1,000 dilution, and ascites fluid or serum from hyperimmunized animals at 1 1,000-1 100,000 dilution. Secondary antibodies are used at dilutions ranging from 1 2,000 to 1 10,000... 

Halofuginone can be also analyzed in chicken serum by a competitive ELISA developed on the basis of monoclonal antibodies (99). In this study, a serum matrix effect that afforded a higher sensitivity for the detection of halofuginone in chicken serum than in assay buffer or in highly diluted serum was observed. The sensitivity of the ELISA improved when used in more concentrated serum. 

Bartelds GM, Wijbrandts CA, Nurmohamed MT, Stapel S, et al. 2007. Clinical response to adal-imumab Relationship to anti-adalimumab antibodies and serum adalimumab concentrations in rheumatoid arthritis. Ann Rheum Dis. 66 921-926. 

The mixture is injected once a week for 3 weeks, and then the animal is maintained for 3 weeks without additional injections. Approximately 25-40 ml of blood is removed from the animal s ear vein to test for antibodies. One week after the first bleed the rabbit is boosted with half the antigen amount used earlier along with incomplete adjuvant incomplete adjuvant does not contain the mycobacteria. Serum is again removed 2 weeks after this injection and tested for antibody response. The enzyme-linked immunosorbent assay is used to determine if the titer (or antibody concentration) of the serum is sufficiently high to establish antibody binding to the antigen. This 3-week cycle is repeated as long as necessary to obtain the antibodies desired. 

Rauer S, Andreou I. Tumor progression and serum anti-HuD antibody concentration in patients with paraneoplastic neurological syndromes. Eur Neurol 2002 47(4) 189-195. 

Figure 8-10. Comparison of serum antibody concentrations following the first and second injections of immunogen (indicated as arrows beneath the abscissa). Note the logarithmic scale for antibody concentration. Time units are unspecified to indicate the great variability encountered with different immunogens. (From B. D. Davis, R. Dulbecco, H. N. Eisen, H. S. Ginsberg, and W. B. Wood, Jr., Microbiology, 2nd ed.. Harper and Row, New York, 1973.)...

Fig. 8. The same antigen solution (horse serum albumin) was placed in the upper layer of these two simple diffusion tubes. The antiserum used in the right-hand tube was that of the left-hand tube (i.e., anti-horse serum albumin) that had been partially absorbed by the homologous antigen, so that not only the antibody concentration had been decreased, but the proportion of precipitating to nonprecipitating antibodies had also been greatly decreased hence the difference in the appeiu-ance of the precipitation zone smaller density and fuzzy (instead of sharp) leading edge. The antiserum in the left-hand tube had been diluted in normal rabbit serum in order to ensure a similar penetration h in the two tubes. From J. Oudin, in Methods in Immunology and Immunochemistry (C. A. Williams and M. W. Chase, eds.), Vol. 3, p. 125. Academic Press, New York, 1971.

Dilutions of the antisera are generally made in 2-fold steps. If the initial dilution to be tested is less than 1 1000 it is advisable to use separate pipettes for each step because antibody from the more concentrated serum carried on the outside of the pipette can introduce serious errors. An initial 1 1000 dilution is most readily made in three successive steps of 10-fold dilutions. 

Add the primary antibody for pAb 1801 in the dilution 1/100 containing 2% bovine serum albumin (BSA) for 30 min (Optimal antibody concentration has to be determined on a section with a proven p53 mutation and positive immunohis-... 

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