PMID- 2948110
OWN - NLM
STAT- MEDLINE
DCOM- 19870127
LR  - 20190824
IS  - 0161-5890 (Print)
IS  - 0161-5890 (Linking)
VI  - 23
IP  - 7
DP  - 1986 Jul
TI  - Cross-linking of IgE-receptor complexes at the cell surface: a fluorescence 
      method for studying the binding of monovalent and bivalent haptens to IgE.
PG  - 769-81
AB  - 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.
FAU - Erickson, J
AU  - Erickson J
FAU - Kane, P
AU  - Kane P
FAU - Goldstein, B
AU  - Goldstein B
FAU - Holowka, D
AU  - Holowka D
FAU - Baird, B
AU  - Baird B
LA  - eng
GR  - AI18306/AI/NIAID NIH HHS/United States
GR  - AI18610/AI/NIAID NIH HHS/United States
GR  - GM35556/GM/NIGMS NIH HHS/United States
GR  - etc.
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Mol Immunol
JT  - Molecular immunology
JID - 7905289
RN  - 0 (Dinitrobenzenes)
RN  - 0 (Dinitrophenols)
RN  - 0 (Fluoresceins)
RN  - 0 (Haptens)
RN  - 0 (Immunoglobulin Fab Fragments)
RN  - 0 (Receptors, Fc)
RN  - 0 (Receptors, IgE)
RN  - 0 (Receptors, Immunologic)
RN  - 0 (Thiocyanates)
RN  - 104077-24-9 (2,4-dinitrophenylaminocaproyltyrosine)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - 42HK56048U (Tyrosine)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
RN  - Q13SKS21MN (2,4-Dinitrophenol)
SB  - IM
MH  - 2,4-Dinitrophenol
MH  - Animals
MH  - Antibody Affinity
MH  - Antigen-Antibody Reactions
MH  - Basophils/*immunology
MH  - Dinitrobenzenes/immunology
MH  - Dinitrophenols/immunology
MH  - Fluorescein-5-isothiocyanate
MH  - Fluoresceins
MH  - Haptens/*immunology
MH  - Immunoglobulin E/*immunology
MH  - Immunoglobulin Fab Fragments/immunology
MH  - Leukemia, Experimental/immunology
MH  - Mast Cells/*immunology
MH  - Mice
MH  - Receptors, Fc/*immunology
MH  - Receptors, IgE
MH  - Receptors, Immunologic/*immunology
MH  - Thiocyanates
MH  - Tyrosine/analogs & derivatives/immunology
EDAT- 1986/07/01 00:00
MHDA- 1986/07/01 00:01
CRDT- 1986/07/01 00:00
PHST- 1986/07/01 00:00 [pubmed]
PHST- 1986/07/01 00:01 [medline]
PHST- 1986/07/01 00:00 [entrez]
AID - 10.1016/0161-5890(86)90089-1 [doi]
PST - ppublish
SO  - Mol Immunol. 1986 Jul;23(7):769-81. doi: 10.1016/0161-5890(86)90089-1.