Monoclonal sites

A two-site immunometric assay of undecapeptide substance P (SP) has been developed. This assay is based on the use of two different antibodies specifically directed against the N- and C-terminal parts of the peptide (95). Affinity-purified polyclonal antibodies raised against the six amino-terminal residues of the molecule were used as capture antibodies. A monoclonal antibody directed against the carboxy terminal part of substance P (SP), covalently coupled to the enzyme acetylcholinesterase, was used as the tracer antibody. The assay is very sensitive, having a detection limit close to 3 pg/mL. The assay is fiiUy specific for SP because cross-reactivity coefficients between 0.01% were observed with other tachykinins, SP derivatives, and SP fragments. The assay can be used to measure the SP content of rat brain extracts. 

When an antigen is injected into an animal, the resulting antibodies are polyclonal, being synthesized by a mixture of B cells. Polyclonal antibodies are directed against a number of different sites (epitopes or determinants) on the antigen and thus are not monospecific. However, by means of a method developed by Kohler and Milstein, large amounts of a single monoclonal antibody specific for one epitope can be obtained. 

Tomlinson E. Davis S.S. (eds) (1986) Site Specific Drug Delivery. Chichester John Wiley. (This deals in part with monoclonal antibodies.)... 

Figure 1. Transverse section of barley leaf epidermal cells taken perpendicular to the long axis of the cells and anticlinal to the leaf surface. The section has been labeled by the EMSIL technique (see Methods) utilizing purified C. sativus endopolygalacturonase and monoclonal antibody EPG-4, which is specific for this enzyme, in order to localize the substrate of the enzyme at the typical site penetrated by the fungal pathogen. Bar = 1 pm. Inset Comparable cell wall region as in Fig. 1 but labeled with monoclonal antibody JIM 5 to localize non-esterified pectin. Bar = 1 pm. Note the identical labeling patterns obtained with either method.

Information about the putative folding of the H,K-ATPase catalytic subunit through the membrane has been obtained by the combined use of hydropathy analysis according to the criteria of Kyte and Doolittle [51], identification of sites sensitive to chemical modification [46,48,50,52-55], and localization of epitopes of monoclonal antibodies [56]. The model of the H,K-ATPase catalytic subunit (Fig. 1) which has emerged from these studies shows ten transmembrane segments and contains cytosolic N- and C-termini [53]. This secondary structure of the catalytic subunit is probably a common feature of the catalytic subunits of P-type ATPases, since evidence supporting a ten a-helical model with cytosolic N- and C-termini has also been published recently for both Ca-ATPase of the sarcoplasmic reticulum and Na,K-ATPase [57-59]. 

The approximate location of the epitopes for more than 40 monoclonal anti-ATPase antibodies has been mapped to various regions within the cytoplasmic domain of the Ca " -ATPase [285,302-304]. All antibodies were found to bind with high affinity to denatured Ca -ATPase, but the binding to the native enzyme showed significant differences depending on the location of antigenic sites within the ATPase molecule. 

N-donor groups in the form of e-amino groups of lysine as nonspecific binding sites for technetium may play an undesired role in Tc-labelling of monoclonal antibodies for tumour imaging [51]. Nonspecifically bound Tc has a poor in vivo stability and appears to increase the undesired liver uptake and reduces tumour uptake. The metal oxidation state and coordination is not yet known. 

The identity of TES-32 and CTL-1 was confirmed by polyclonal antibodies to recombinant CTL-1, which bound to native TES-32, and by monoclonal antibody Tcn-3, raised to native TES-32 (Maizels et al, 1987), which specifically recognized recombinant CTL-1. The CTL-1 sequence also contained three sites for Afglycosylation, which had previously been shown to be present on TES-32 (Page and Maizels, 1992). Both Tcn-3 and polyclonal antibody to the recombinant CTL-1 protein localize to the cuticle of the infective larvae by immunoelectron microscopy (Fig. 12.2). 

Rituximab -monoclonal antibody to CD20 (B-cell surface antigen) -fever, chills, malaise -nausea, vomiting -flushing -bronchospasm, angioedema, urticaria -rhinitis -pain at disease sites -tumor lysis syndrome may occur in patients with high peripheral lymphocyte count... 

The conformation and orientation of adsorbed proteins has been examined with monoclonal antibodies that recognize a specific site in a protein of interest. Keselowsky et al. examined the conformation of Fn adsorbed to SAMs that carried methyl, hydroxyl, carboxyl, and amine groups [79]. They used monoclonal antibodies that recognized the central cell-binding domain of Fn near the RGD motif. Different SAM functionalities differentially modulated the binding affinities of the monoclonal antibodies (OH > COOH = NH2 > CH3). The strength of cell adhesion to these... 

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