Abstract:
:We have developed a method for use in investigating factors controlling the binding and cross-linking by bivalent haptens of immunoglobulin E (IgE) bound to receptors on rat basophilic leukemia (RBL) cells. This method employs monoclonal anti-2,4-dinitrophenyl (DNP) IgE that is labeled with fluorescein-5-isothiocyanate (FITC), and it measures FITC quenching that accompanies DNP occupation of the antibody combining sites in a titration experiment. The validity of this approach is demonstrated using the monovalent hapten DNP-L-lysine. The affinity constant for this ligand obtained by the FITC quenching method is compared with those obtained with previously established methods: equilibrium dialysis and quenching of endogenous tryptophan for IgE in solution and [3H]-DNP-L-lysine binding to IgE on cells. The FITC quenching method has been used to carry out a detailed study of the binding of monovalent DNP-aminocapryol-L-tyrosine (DCT) and bivalent (DCT)2-cystine to FITC-IgE and its Fab fragments in solution. Intrinsic (K) and cross-linking (Kx) affinity constants are obtained by analyzing the binding curves in terms of simple equilibrium equations. With these DCT haptens the ability of this method to assess hapten binding and cross-linking of IgE bound to receptors on RBL cells is shown.
journal_name
Mol Immunoljournal_title
Molecular immunologyauthors
Erickson J,Kane P,Goldstein B,Holowka D,Baird Bdoi
10.1016/0161-5890(86)90089-1subject
Has Abstractpub_date
1986-07-01 00:00:00pages
769-81issue
7eissn
0161-5890issn
1872-9142journal_volume
23pub_type
杂志文章abstract::Structurally diverse dextrans from Leuconostoc mesenteroides and related bacteria have been used extensively in fundamental immunochemical studies such as induction and characterization of anti-dextran antibodies, as well as in studies of their interaction with pneumococcal antisera, normal bovine serum, concanavalin ...
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pub_type: 杂志文章
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