The characterization and adsorption of sensitizing antibodies (original) (raw)

A method of measuring adsorption without washing the tissue has been devised and studied.

Diffusion through chopped lung tissue has been studied from a theoretical and experimental point of view. Predicted curves for the y-globulin content of the extracellular space have been calculated and applied to experimental results. The rate of uptake was found to be at least partly diffusion controlled. Adsorption equilibrium and rate constants have been rigorously defined. No evidence was found for a fast initial phase of sensitization or adsorption.

Calculations have been performed concerning the consequences of slow steady y-globulin uptake after long periods of incubation.

The amounts of rabbit y-globulin adsorbed onto lung tissue were found to be of the same order of magnitude, or somewhat lower, than those previously reported.

No evidence against a linear adsorption isotherm was found in any experiments.

Neither adsorption nor sensitization were altered by reducing the calcium concentration during passive sensitization.

Reduction of the ionic strength of the medium caused a large increase in the amount of Y-globulin adsorbed but did not increase sensitization.

Two guinea pig antibodies have been separated by preparative electrophoresis and ion exchange chromatography.

The antigenic relationship and purity of γ₁-and γ₂-globulins have been studied. No contamination was detectable in γ₂_globulin, but y₁-globulin contained fast γ₂-globulin and sometimes ß-globulins as well.

Ovalbumin (crystallized 5 times) has been shown to contain at least four proteins.

It has been shown that γ₁-globalin anti-bodies sensitize lung tissues in very low concentrations, but no evidence was found that the very low sensitizing power of γ₂-globulin antibodies was not due to impurity.

Quantitative passive cutaneous anaphylaxis experiments performed in parallel with the tests on lung tissue showed that whenever the γ₂-globulin fraction contained antibody it appeared more potent relative to γ₂-globulin than when tested on lung tissue although the γ₁-globulin was always considerably more potent in both tests.

The skin sensitization produced by the γ₂--globulin fraction disappeared faster than that produced by the γ₁-globulin fraction.

It was not possible to detect enough γ₁-globulin contaminant in the γ₂-globulin fraction to account for the skin sensitizing ability of the latter.

It was concluded that γ₂-globulin antibody must have some skin sensitizing ability of its own, but that it is considerably less potent than the γ₁-globulin antibody.

No difference was detectable between the extents of adsorption of γ₁-and γ₂-globulins onto lung tissue.

An equation has been derived describing the loss of thiosulphate by radiative oxidation in iodine-131 solutions.

An analysis has been presented of interpolation and other errors in a rapid method for assaying large numbers of histamine solutions using visual linear interpolation between two standards.