Antibody panning

Cetuximab Herceptin therapeutic monoclonal antibody Panning on plastic-immobilized protein... 

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. 

Davies, J., Jiang, L., Pan, L.-Z., LaBarre, M., Anderson, D., and Reff, M., Expression of GnTlIf in a recombinant anti-CD20 CHO production cell line expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FcyRlll, Biotechnol. Bioeng., 74, 288-294, 2001. 

While invertebrate tropomyosins are likely pan-allergens, vertebrate tropomyosins appear to be nonallergenic (Reese et ah, 1999). Using bioinformatics approaches to compare the sequences of tropomyosins from various species, Goodman et ah (2002) determined that tropomyosins from vertebrate species — rabbit, pig, chicken, and human — share 53-57% amino acid sequence identity to the known shrimp tropomyosin allergen. Met e 1. This comparison likely explains why vertebrate tropomyosins are not allergenic and do not cross-react with IgE antibodies specific to invertebrate tropomyosins. 

Tissue specimens are snap-frozen in liquid nitrogen for 30 sec immediately after removal and then transferred to a cryostat (Kammerer et al., 2001). Serial frozen sections of 5 pan thickness are cut and placed on silane-coated slides. They are air-dried for 30 sec, fixed in acetone for 1 min at room temperature (22°C), and air-dried at 22°C (Fig. 6.11). The sections are incubated with primary antibody in the antibody diluent (Dako) for 3 min by placing the slide horizontally on a hot plate at 37°C. (All incubation steps are carried out by placing the slide horizontally on the hot plate at 37°C.) Following a brief rinse in TBS, the sections are incubated with the goat-anti-mouse EnVision-HRP-enzyme conjugate for 3min at 37°C. 

Hawlisch, H., Muller, M., Frank, R., Bautsch, W., Klos, A., and Kohl, J. (2001) Sitespecific anti-C3a receptor single-chain antibodies selected by differential panning on cellulose sheets. Anal. Biochem. 293, 142-145. 

For some antibodies, positive clones were found even after the first round of panning. It seems that further enrichment is undesirable in these cases because there is the possibility that this might lead to isolation of fewer clones with higher affinity for the antibodies. 

Although T cells can be divided into two populations (helper/inducer and cytotoxic/suppressor) based on CD4 and CD8 expression, T cells can also be divided into two populations based on the structure of the antigen receptor complex, either alp, or yIS, associated with the CD3 antigen (DIO). The yIS T cells represent approximately 5% of all T cells, lack expression of CD5, CD4, and often CD8, are thought to be associated with a more primitive type of immune response, and are usually found in the splenic red pulp and epithelial sites (A15, C26, T6). The alj3 T cells usually express all four pan-T cell antigens (CD2, CD3, CD5, and CD7) and either CD4 or CD8. The CD4-positive cells can be further divided based on cytokine secretion Thl cells, which secrete interleukin-2 (IL-2) and interferon y and help macrophages and other T cells, and Th2 cells, which secrete IL-4, IL-5, IL-6, and IL-10 and help B cells in antibody production (DIO, Dll). 

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