Search For Specific Antibodies Research Articles

Phage display of proteins.

In recent years the phage display approach has become an increasingly popular method in protein research. This method enables the presentation of large peptide and protein libraries on the surface of phage particles from which molecules of desired functional property(ies) can be rapidly selected. The great advantage of this method is a direct linkage between an observed phenotype and encapsulated genotype, which allows fast determination of selected sequences. The phage display approach is a powerful tool in generating highly potent biomolecules, including: search for specific antibodies, determining enzyme specificity, exploring protein-protein and protein-DNA interactions, minimizing proteins, introducing new functions into different protein scaffolds, and searching sequence space of protein folding. In this article many examples are given to illustrate that this technique can be used in different fields of protein science. The phage display has a potential of the natural evolution and its possibilities are far beyond rational prediction. Assuming that we can design the selection agents and conditions we should be able to engineer any desired protein function or feature.

MH27 Binds to Several Types of Epithelial Junctions by Em-Immuncytochemistry in the Nematode C. Elegans

The monoclonal antibody MH27 was used to test a post-embedding method for EM immunocytochemistry in the nematode C. elegans. Primary antibody localization was performed on LR Gold thin sections of lightly fixed him-5 adult animals, followed by a gold-linked anti-mouse IgG secondary antibody (AuroProbe, Amersham Life Science) and viewed on a Philips CM 10 electron microscope. Methods for fixation, embedding, sectioning and antibody procedures followed those of Selkirk et al. (1990) and Hall (1995). Relatively few examples of immunolabelling have been previously demonstrated in C. elegans at the EM level. We have previously found that LR White embedment was unsuitable, as it causes severe shrinkage in nematode tissues. Selkirk and coworkers showed that LR Gold was preferable for EM-immunocytochemistry, and used it to localize several surface antigens. Okamoto and Thomson (1985) previously demonstrated successful results with post-embedding labeling of Epon embedded material, but we anticipate that many antigens will not be well exposed in an Epon thin section.MH27 was generated during a search for specific antibodies to muscle proteins (Francis and Waterston, 1985). The antibody is now widely used for immunofluorescence studies to demarcate the boundaries of the hypodennal cells, and has proven useful to follow cellular development during embryogenesis and larval stages in C. elegans and other nematode species (Fitch and Emmons, 1995). We were interested to learn the ultrastructural localization of the MH27 antibody, as well as to further develop this technique for C. elegans tissues. We used the him-5 strain so that both male and hermaphrodite animals could be sampled.

Comparison of two assays (EIA and EITB) and two samples (saliva and serum) for the diagnosis of neurocysticercosis

The enzyme immunoassay (EIA), standardized with a crude extract, and the enzyme-linked immunoelectrotransfer blot assay (ETIB) with glycoprotein antigens, were compared by using saliva and serum in the search for specific antibodies against Taenia solium larvae, for the diagnosis of neurocy-sticercosis. Saliva was slightly more sensitive in EIA (82·1%) than serum (74·1%). In EITB serum was far more sensitive (100%) than saliva (70·4%). The use of EITB with serum is thus an excellent choice for diagnosis of clinical cases of neurocysticercosis, while EIA using saliva represents a useful combination for diagnosis and, especially, epidemiology, because saliva is easily obtained by a painless and non-invasive procedure and the technique is simpler to perform. Furthermore, cross-reactivity of EIA with Echinococcus does not interfere in countries like Mexico where human hydatid disease is not present.

3] Production and characterization of monoclonal idiotypes and anti-idiotypes for small ligands

The procedures described in this chapter have enabled us to identify and characterize monoclonal antibodies and their respective anti-idiotypes. We have developed several different types of immunoassays which afford greater flexibility to the investigator, depending on the type of antibodies desired and the availability of labeled antigens. Use of the intrasplenic injection technique for the final booster immunization prior to the fusion protocol has enabled us to achieve more consistent results than the usual intravenous or intraperitoneal injection routes. Isoelectric focusing of tissue culture supernatant from monoclonal antibody-secreting clones can easily identify possible duplicate clones, and thereby reduces the amount of labor required for extensive characterization of a large number of clones. We have found that these techniques have enabled us to identify "sister clones" or redundancies in our collection of antiligand and anti-idiotype antibodies rapidly and accurately. These various techniques have allowed us to save much time, labor, and money in the search for specific antibodies with desired characteristics.

Antibodies against pleuropneumonia-like organisms in patients with salpingitis.

Since the first report of Dienes and Edsall (1937), a number of communications have recorded the isolation of pleuropneumonia-like organisms (PPLO) from the genital tract of the human female, especially in association with inflammatory diseases such as bartholinitis, pelvic abscess, and salpingitis (Melen and Odeblad, 195 1; Randall, Stein, and Ayres, 1950; Dienes, Ropes, Smith, Madoff, and Bauer, 1948; Klieneberger-Nobel, 1945, 1959, 1962). In only a few cases has a search for specific antibodies been undertaken. Beveridge, Campbell, and Lind (1946) found antibody in a proportion of the human sera they examined, but were unable to correlate the occurrence of antibody with any clinical condition. Stokes (1955) detected antibody against PPLO in four patients (two with puerperal sepsis, one with pyosalpinx, and one with empyema); PPLO but no other pathogens were isolated from the blood, genital tract, or operation wound. Stokes (1959) reported a further case of infection following panhysterectomy, in which PPLO were isolated from the blood and antibody was present in high titre. Melen and Gotthardson (1955) found high titres of PPLO antibody in five patients with salpingo-oophoritis. Card (1959) examined serologically 56 PPLO strains isolated from the human genital tract and found that they belonged to one broad serological group; she distinguished them serologically from PPLO isolated from the mouth, from sewage, and from various animals. This serological type-specificity of the genital strain made possible a survey of a large number of sera in terms of antigens from a single representative PPLO strain. Sera from patients with venereal infections, non-venereal diseases, healthy blood donors, and children were examined. Female patients from venereal disease clinics had