Monoclonal antibody binding

NH Chiem, DJ Harrison. Monoclonal antibody binding affinity determined by microchip-based capillary electrophoresis. Electrophoresis 19 3040-3044, 1998. 

Monoclonal antibody-binding affinity was determined by microchip-based capillary electrophoresis with LIF detection (33). The mixing was carried out off-chip, and the on-chip separations were performed in less than 60 s (Fig. 9). A Scatchard plot analysis resulted in an affinity constant for the monoclonal anti-BSA antibody to fluorescently labeled BSA (BSA ) of... 

As a result, we started to study the mode of reaction of atypical elements with certain monoclonal antibodies binding to individual antigens, tumor markers specific for the respective cellular populations. Moreover, the method can be used to determine the degree of their maturity, presence of individual receptors, and the state of activation in the course of their cellular cycle. 

C. Aspirin inhibits platelet cyclooxygenase. Abciximab, a monoclonal antibody, binds to and inhibits the platelet glycoprotein Ilb/IIIa receptor. Dipyridamole inhibits platelet cyclic AMP phosphodiesterase and raises cyclic AMP levels. Eptifibatide binds to the glycoprotein Ilb/IIIa complex. 

Zhang, T.-H. Luo, J. Zhou, J.-M. Conformational changes at the active site of adenylate kinase detected using a fluorescent probe and monoclonal antibody binding. Biochimie, 84, 335-339 (2002)... 

Using Antibodies to Probe Structure-Function Relationships in Proteins A monoclonal antibody binds to G-actin but not to F-actin. What does this tell you about the epitope recognized by the antibody ... 

Roe, R., Robins, R. A., Laxton, R R., and Baldwin, R W (1985) Kinetics of divalent monoclonal antibody binding to tumor cell surface antigens using flow cytometry—standardization and mathematical analysis Mol Immunol. 22, 11—21... 

Fig. 3.8 Two types of monoclonal antibody binding. Left Fab-directed binding. Right Fc-directed binding.

Martens, W., Greiser-Wilke, I., Harder, T.C., et al. (1995) Spot synthesis of overlapping peptides on paper membrane supports enables the identification of linear monoclonal antibody binding determinants on morbillivirus phosphoproteins. Vet. Microbiol. 44, 289-298. 

In die proximal linkage immunoassay system, two kinds of monoclonal antibodies that can bind to two different antigenic determinants on one antigen are selected. The selected pairs of monoclonal antibodies are conjugated with the proper pair of reactant molecules (Rx, R2) as shown in Table 7 (S3). When unlabeled antigen is added, each reactant-labeled monoclonal antibody binds to a proximal antigenic site of the antigen and the whole process proceeds. 

Bhart, N.M., Bieber, M.M., Chapman, C.J. Human anti-lipid A monoclonal antibodies bind to human B cells and i-antigen on core red blood cells. J Immunol 151 (1993) 5011-5021. 

Di Padova, F., Brade, H., Barclay, G.R., Poxton, I.R., Liehl, E., Schuetze, E., Kocher, H.P., Ramsay, G., Schreier, M.H., McClelland, D.B.L., Rietschel, E.T. A broadly cross-protective monoclonal antibody binding to Escherichia coli and Salmonella lipopolysaccharide. Infect Immun 61 (1993) 3863-3872. 

In addition to the immunogenic potential of murine monoclonal antibodies, another disadvantage of murine antibodies is their rapid elimination from the circulation relative to human antibodies. Muromomab has an elimation half-life of about 18 hours, whereas human antibodies have half-lives of weeks. Human antibodies have long serum persistence because they are recycled by endothelial cells. The Fc portion of murine monoclonal antibodies binds very poorly to human FcRn, and therefore murine antibodies are not recycled resulting in their poor serum persistence [7]. The converse, however, is not true. Human Fc binds well to murine FcRn, and therefore human antibodies have long serum persistence in mice. The lack of FcRn binding in combination with a HAMA response results in rapid clearance from the serum. 

Recombinant human monoclonal antibody binds to vascular endothelial growth factor... 

Antitumor necrosis factor human monoclonal antibody binds to TNI-a... 

Murine monoclonal antibody binds to the CD3 antigen of T lymphocytes... 

Recombinant humanized monoclonal antibody binds to CD 11a on leukocytes... 

Chimeric monoclonal antibody binds with high affinity to the soluble and transmembrane forms of TNFa and inhibits binding of TNFa with its receptors... 

The conformational energy calculations received support subsequently from two types of experimental study by Smith-Gill et al. (1984). The affinity of ring-necked pheasant lysozyme, in which Asn and Arg at positions 113 and 114 are replaced by Lys and His, respectively, is the same for (NAG)e as that of domestic hen egg-white lysozyme (i.e., the right side is not involved). They also showed that a monoclonal antibody bind-... 

Hudecz, F. and Price, M. R. (1992) Monoclonal antibody binding to peptide epitopes conjugated to synthetic branched polypeptide carriers. Influence of the carrier upon antibody recognition. J. Immunol. Methods 147, 201-210. 

Fig. 7. Negative-stained images of monoclonal antibodies binding to apoBlOO on LDL. (A) An equimolar mixture of LDL and each of two anti-apoBlOO monoclonal antibodies, MB47 and MB 19, were adsorbed to the carbon-coated grid, stained with 1% uranyl acetate, and examined in the electron microscope. The insert shows both open (o) and closed (c) complexes. Bar = 0.1 p.m. (B) Complexes formed between LDL and monoclonal antibodies MB47 and MB24. (From Chatterton etal., 1991.)...

Fig. 9. Observed angular distributions of pairs of monoclonal antibodies binding to LDL. (A) The distribution of 124 angles measured between 4G3 and B3 yield an average angle of 104°. (B) The distribution of 139 angles measured 4G3 and B4 yield an average angle of 60°.

M. A. Schenerman and T. J. Collins, Determination of a monoclonal antibody binding activity using immunodetection, Anal. Biochem. 217(2) 241 (1994). 

A possible drawback to the use of antibodies for the treatment of toxicity is the possibility of anti-idiotypes being produced against the monoclonal antibody binding site. These antibodies could potentially mimic drug-like structural features necessary for binding to a receptor. However, antibodies do not cross the blood-brain barrier in significant amounts and studies to date have not found this to be a problem, especially when Fab fragments are used. 

Another interesting application is the analysis of the specificity of monoclonal antibodies, i.e., whether they recognize different epitopes on the antigen (Friguet et al., 1983). After coating the antigen, two monoclonal antibodies are added, either to separate or to the same well and the amount of bound antibody is quantitated with enzyme-labeled anti-mouse IgG. Additivity of enzyme activity is observed when the monoclonal antibodies bind to distinct epitopes. The additivity index (A/) for a pair of antibodies is then defined as ... 

Benzo[a]pyrene (62.5 or 500 pg) was applied once to the shaved backs of male C57BL/6 mice (Bjelogrlic et al. 1994). Mice were killed at different time intervals after the treatment. DNA was isolated from the skin, purified, and analyzed for benzo[a]pyrene-7, 8-diol-9, 10-epoxide-DNA adducts. Skin was also evaluated for monoclonal antibody binding to mouse p53 protein, which has been shown to increase in response to DNA damage. Alterations in p53 are the most frequently observed mutations in human cancer. Benzo[a]pyrene-7,8-diol-9, 10-epoxide-DNA adducts reached their maximum concentration 24 hours after the treatment, and decreased sharply within 1 week, regardless of the dose. An increase in p53 protein was seen only after treatment with 500 pg benzo[a]pyrene. 

Specificity of Anti-thymidylate Synthase Monoclonal Antibodies Binding on Western blots... 

We have developed ELISA systems using a polyclonal (12) and a monoclonal antibody (K4) to Mn-SOD. The Mn-SOD is composed of four identical subunits therefore, one can use a single monoclonal antibody to bind and detect the Mn-SOD. Namely, a monoclonal antibody binds one molecule of the tetrameric enzyme and then the same type of monoclonal antibody linked with peroxidase binds to another monomer of the enzyme. A schematic representation of the ELISA is shown in Fig. 1. 

Ellgaard, L., Holtet, T.L., Moestrup, S.K., Etzerodt, M. and Thogersen, H.C. (1995). Nested sets of protein fragments and their use in epitope mapping characterization of the epitope for the S4D5 monoclonal antibody binding to receptor associated protein. J. Immunol. Meth. 180, 53-61. 

Jensen, T.H., Jensen, J., Szilvay, A.M. and Kjems, J. (1997). Probing the structure of HIV-1 Rev by protein footprinting of multiple monoclonal antibody binding sites. FEBS Lett. 414, 50-54. 

---