Antibodies using

The potential of B lymphocyte depletion as an approach to therapy has been confirmed in RA patients seropositive for rheumatoid factor and/or anti-CCP antibodies using the anti-CD20 mAb, rituximab. 

To allow all culture productiou to be coutrolled, a method for rapid analysis is required. Prior to development of an LC-MS method, the analysis was both complex and time-consuming, involving the purification of a relatively large amount of the antibody using affinity chromatography, enzymatic release, and subsequent derivatizafion of the oligosaccharides and their analysis by using capillary electrophoresis. 

IgG below specified limit (if monoclonal antibodies used in purification) ELISA or RIA... 

Santora, L. C., Krull, I. S., and Grant, K., Characterization of recombinant human monoclonal tissue necrosis factor-alpha antibody using cation-ex-change HPLC and capillary isoelectric focusing, Anal. Biochem., 275, 98,1999. 

Differences in the relative proportion of f-PSA and PSA-ACT can affect the result obtained for t-PSA because of the differences in the nature of calibration and the molar response, sensitivity, and specificity of antibodies used in various immunoassays. The efficiency of these immunoassays has been evaluated by several investigators. Because the proportion of free and complexed PSA varies in benign and malignant diseases, these immunoassays measure one form or the other, giving rise to different results for different patient groups. It is very important that data from clinical studies support the proposed intended uses of these assays, since as many as 5 percent of men with a negative free PSA test (free PSA values >25 percent) will have cancer and not be recommended for biopsy. Therefore, a goal for standardization is to detect total and free PSA accurately in equimolar fractions. 

Serological techniques can detect target bacteria rapidly in mixtures, but their accuracy depends on the specificity of the antibody used. The use of monoclonal instead of polyclonal antibodies may increase specificity.49,52,58 However, because the same epitope can be present in more than one species, a monoclonal antibody against one species may cross-react with other bacteria.50 For this reason serological methods are not always successful for detection of bacteria in environmental samples and nucleic acid-based methods are now commonly used. 

Our laboratory is interested in the biology of mammalian SGs and PBs, and in understanding their roles in the spatial regulation of mRNA translation and decay. The methods and procedures presented here describe our present knowledge of SG and PB assembly and composition. We indicate some commercially available antibodies useful as SG and PB markers, describe some immunocytochemical protocols that we employ, and offer some caveats regarding data interpretation. 

Pan CC, Chen PC, Tsay SH, et al. Cytoplasmic immunoreactivity for thyroid transcription factor-1 in hepatocellular carcinoma a comparative immunohistochemical analysis of four commercial antibodies using a tissue array technique. Am. J. Clin. Pathol. 2004 121 343-349. 

The IHC stain procedure is a multistep staining protocol, the various steps intended to provide amplification of stain results. Therefore, a control system must include elements to control each step of the stain process. Such a control should also include a range of reactivities, and that range ideally would encompass the total expression range expected for the measured component. The control should also monitor each step of the multistep protocol. This author has devoted a number of years to this concept, resulting in a patented control for multistep staining processes.14 Such a control provides sufficient information to monitor every IHC stain run, and when the control is evaluated quantitatively, normalization of data from one stain run to another within the same laboratory, and even between laboratories. A process control is a measure of the stain protocol and does not take the place of a control for the primary antibody. While the primary antibody control should include range of expression level detection, a different primary control must be present for every primary antibody used in a stain run (Fig. 10.4). 

It became apparent very early in the development of such agents that their conception and design was much easier to imagine than to successfully implement. Monoclonal antibodies used... 

Purify the reduced antibody using gel filtration on a column of Sephadex G-25. Concentrate the protein to lOmg/ml using centrifugal concentrators. 

Figure 22.21 Antibodies may be conjugated to liposomes using an indirect approach incorporating a (strept)avidin-biotin system. Biotinylated liposomes may be complexed with biotinylated antibodies using (strept)avidin as a bridging molecule or may be complexed with an antibody-(strept)avidin conjugate.

Goldman, E.R. et al. (2002b) Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays. Anal. Chem. 74, 841-847. 

Yoshitake, S., Yamada, Y., Ishikawa, E., and Masseyeff, R. (1979) Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N-(4-carboxycyclohexyl methyljmaleimide. Eur.J. Biochem. 101, 395-399. 

Zopf, D.A., Smith, D.F., Drzeniek, Z., Tsai, C.-M., and Ginsburg, V. (1978a) Affinity purification of antibodies using oligosaccharide-phenethylamine derivatives coupled to Sepharose. In Methods in Enzymology, (V. Ginsburg, ed.), Vol. 50, pp. 171-175. Academic Press, New York. 

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