Activation of human complement by mouse and mouse/human chimeric monoclonal antibodies.

The complement (C)-activating capabilities in human serum of 32 mouse and 10 mouse/human chimeric MoAbs of different isotypes, and their fragments, were tested in vitro. Activation of C via the classical pathway (CP) was performed in 1% factor D-deficient serum in gelatin containing Veronal buffer in the presence of calcium and magnesium (GVB++), while activation of the alternative pathway of C (AP) was assessed in 10% C1q-depleted serum in the presence of 5 mM MgCl2 in GVB++. The C-activating ability of MoAbs was expressed relative to the degree of activation of complement by aggregated IgG for the CP and relative to mouse IgG1 for the AP.

Modulation of the structure-binding relationships of antagonists for muscarinic acetylcholine receptor subtypes.

1. Membranes from rat cerebral cortex, myocardium and extraorbital lacrimal gland were used as sources of M1, M2 and M3 muscarinic acetylcholine receptors respectively and the affinities of seven antagonists for the three subtypes were examined under different experimental conditions. 2.

The modes of binding of ligands to cardiac muscarinic receptors.

Ionizable groups on the cardiac M2 muscarinic receptor which regulate the binding of ligands have been examined by studying the pH dependence of the ligand affinity constants. The presence of three titratable residues (approximate pK values, 5.4, 6.

Binding and hydrodynamic properties of muscarinic receptor subtypes solubilized in 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propanesulfonate.

The muscarinic receptor from the cerebral cortex, heart, and lacrimal gland can be solubilized in the zwitterionic detergent 3-(3-cholamidopropyl)dimethylammonio-2-hydroxy-1-propane sulfonate (CHAPSO) with retention of high affinity [3H]N-methyls-copolamine binding. However, in this detergent there are significant differences in the binding properties of the receptors, compared with those observed in membranes and digitonin solution. Some agents retain a degree of selectivity.

The existence of stable enantiomers of telenzepine and their stereoselective interaction with muscarinic receptor subtypes.

[3H]Telenzepine has been shown to bind with high affinity (3 x 10(9) M-1) to a subpopulation of muscarinic binding sites in rat cerebral cortex, which have a high affinity for pirenzepine. The binding kinetics were very slow at 30 degrees. Only 50% of the [3H] telenzepine was found to be capable of binding to the receptors with high affinity.