Monoclonal antibodies characterization

Urmann, M., Graalfs, H., Joehnck, M., Jacob, L.R., and Freeh F Ch. (2010) Cation-exchange chromatography of monoclonal antibodies -characterization of a novel stationary phase designed for production-scale purification. mAbs, 2, 395-404. 

Visilizumab is a humanized anti-CD3 monoclonal antibody characterized by a mutated IgG2 isotype, lack of binding to Fey receptors, and an ability to induce apoptosis selectively in activated T cells. 

The investigation and characterization of cell surfaces by probing with monoclonal antibodies is one of the most vital areas of application. In this context they have been used in the following ways ... 

Xiang SH, Doka N, Choudhary RK, Sodroski J, Robinson JE. Characterization of CD4-induced epitopes on the HIV type 1 gpl20 envelope glycoprotein recognized by neutralizing human monoclonal antibodies. AIDS Res Hum Retroviruses 2002 18(16) 1207—1217. 

Parasitism by T. spiralis has been a subject of scientific interest for over 150 years. Recently, considerable attention has been paid to the parasite by immunologists interested in immunity to nematodes in general, and mucosal immunity in particular. It has been shown that glycan-specific antibodies are highly effective mediators of host defence against intestinal 7. spiralis infection. Protective monoclonal antibodies have been used to elucidate mechanisms of worm expulsion, as well as to identify molecules that the parasite uses to create its niche. In the future, detailed characterization of these molecules and their functions should afford additional insights into parasitism by Trichinella spiralis, and possibly also by other types of pathogen. 

The involvement of mast cells in host protection against nematode infection is well characterized in T. spiralis infection. W/Wv mice exhibited a significant delay in worm expulsion, and treatment with either anti-SCF or anti-SCF receptor monoclonal antibody dramatically inhibited mast cell responses and expulsion of T. spiralis for the duration of treatment (Donaldson et al., 1996). W/Wv mice also lack interstitial cells of cajal and intraepithelial y T cells (Maeda et al., 1992 Puddington et al., 1994), which may contribute to the impaired response in these animals (see below). However, supporting evidence of a role for mast cells in protection against T. spiralis comes from studies in which overexpression of IL-9 in mice (which is known to influence the mast cell responses see above) resulted in an extremely rapid mast cell-dependent expulsion of T. spiralis (Faulkner et al., 1997). 

Yasuda K, Yamashita S, Aiso S, et al. Immunohistochemical study of gamma-glutamyl transpeptidase with monocloncal antibodies. I. Preparation and characterization of monoclonal antibodies to gamma-glutamyl transpeptidase. Acta Histochem. Cytochem. 1986 19 589-592. 

The simplicity of this assay makes it highly attractive for further development. Some of the limitations include the need for the preparation of relatively large amounts of appropriately modified DNA for the initial immunization. This may be reduced with the development of in vitro immunization techniques in which as little as 5ng of antigen can be effective. The antibodies must also be characterized once prepared. For example, a variety of monoclonal antibodies against B[a]PDE-modified DNA have been prepared (42) which show varying specificity. At one extreme, some require the full structure of the adduct bound to DNA, at the other, BtalP tetraol will effectively compete. What is not clear at the moment is how specific such antisera are for a particular PAH. Will these antisera recognize only B[a]P tetraol structures or those of any diol epoxide modified DNA ... 

There are literally hundreds of markers that are currently available for the mouse and human than can be used to characterize lymphoid and myeloid cells and subsets in primary and secondary lymphoid organs. Many of the markers expressed in mammals are highly conserved across species and have been designated as genetic clusters of differentiation (CD). CDs can be identified with fluorescently labeled monoclonal antibodies. As presented previously, when combined with other fluorescent probes, important information on intracellular biochemistry and cell function can be obtained. Many of the biochemical markers used by immunotoxicologists are common to other... 

Funke, C. et al., Harbor seal (Phoca vitulina) C-reactive protein (C-RP) purification, characterization of specific monoclonal antibodies and development of an immuno-assay to measure serum C-RP concentrations, Vet. Immunol. Immunopathol., 59, 151, 1997. 

De Guise, S. et al., Characterization of a monoclonal antibody that recognizes a lymphocyte surface antigen for the cetacean homologue to CD45R, Immunology, 94, 207, 1998. 

Bartek J, Vojtesek B, Staskova Z, Bartkova J, Kerekes Z, Rejthar A, Kovarik J (1991) A series of 14 new monoclonal antibodies to keratins characterization and value in diagnostic histopa thology. J Pathol 164(3) 215 224... 

L7. Lafferty, M. A., Salamon, A. M., and Usher, D. C., Immunochemistry of human Lp(a) Characterization of monoclonal antibodies that cross-react strongly with plasminogen. J. Lipid Res. 32, 277-292 (1991). 

R. Bethell, M. Dawson, and M. J LaFoe, Characterization of monoclonal antibodies to cell surface antigens by particle concentration fluorescence immunoassay (PCFIA), BioTechniques 3, 466-473 (1985). 

Fischer D, Kissel T (2001) Histochemical characterization of primary capillary endothehal cells from porcine brains using monoclonal antibodies and fluorescein isothiocyanate-labelled lectins Implications for drug delivery. Europ J Pharm Biopharm 52 1-11... 

ACE has been used to characterize protein-sugar interactions,45 DNA binding to an anti-DNA monoclonal antibody,46 antibody-antigen47 antisteroidal inflammatory drugs,48 and prion protein in sheep brain preparations.49... 

Roberts G.D., Johnson W.P., Burman S., Anumula K.R., and Carr S.A. (1995), An integrated strategy for structural characterization of the protein and carbohydrate components of monoclonal antibodies application to anti-respiratory syncytial virus MAb, Anal. Chem. 67, 3613-3625. 

A., and Roepstorff P. (2001), Characterization of a recombinant monoclonal antibody by mass spectrometry combined with bquid chromatography. J. Chromatogr. 

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