Separation of antigens by immunological specificity: Use of disulphide-linked antibodies as immunosorbents

1. γ-Globulin concentrates of antisera prepared against ovalbumin and human serum albumin were thiolated and cross-linked to form insoluble polymers. 2. These immunosorbents were of low solubility and of high capacity for homologous antigen. 3. The high specificity of these immunosorbents was demonstrated by fractionation of various binary mixtures of fluorescent ovalbumin, ¹³¹I-labelled human serum albumin, lysozyme and ribonuclease.     

Separation of antigens by immunological specificity: Use of cellulose-linked antibodies as immunosorbents

1. Immunosorbents were prepared by coupling activated aminocellulose with the γ-globulin concentrates of antisera prepared against ovalbumin and human serum albumin. 2. The immunosorbents were low in solubility, but high in capacity for homologous antigens. 3. The high specificity of these immunosorbents was demonstrated by their use in fractionating various mixtures of fluorescent ovalbumin, ¹³¹I-labelled human serum albumin, lysozyme and ribonuclease.     

Studies on the purification of chlamydial agents grown in yolk sacs of embryonated eggs using disulphide-linked immunosorbents and enzymes.

Immunosorbents were derived from avid and non-avid sera raised in rabbits to multiple or single injections of chlamydiae passaged once or three times in HeLa cells after routine passage in eggs. Egg-derived suspensions of chlamydiae required pretreatment before application to immunosorbent columns; this was most conveniently done by fractionation on Sepharose 4B. Immunosorbents derived from avid serum had greater capacity than those from non-avid sera. However, organisms were desorbed in low yield from an avid immunosorbent, but in higher yields from non-avid immunosorbents. Even under the best conditions, the immunosorbents yielded suspensions of organisms still contaminated with egg antigens. Partially purified suspensions of chlamydiae were also treated with phospholipase C to yield suspensions of high purity.     

Regulation of in vivo immunological reactions by monoclonal antibody against lymphocyte activation antigen

In vivo effects of a monoclonal antibody that recognizes rat lymphocyte activation antigen were studied. Spleen cells obtained from sheep red blood cell (SRBC)-immunized rats developed strong PFC response against SRBC. However, the 5C6-F4 treatment resulted in the inhibition of subsequent development of PFC response. The suppression of PFC response was due to the inhibition of generation of helper T cells, but not due to the preferential induction of suppressor cells. In addition, 5C6-F4 antibody was also found to inhibit the clinical expression of collagen-induced rat arthritis and the synovial inflammation in collagen-induced arthritis rats. Furthermore, the in vivo generation of cytotoxic cells against syngeneic tumor cells was also inhibited by 5C6-F4 antibody. The in vivo administration of 5C6-F4 antibody did not cause any pathological changes in brain, lung, liver, kidney, spleen, thymus, and lymph nodes.     

Separation of Dirofilaria immitis allergen from the IgG-inducing antigens.

Allergen in crude extract of Dirofilaria immitis was purified and separated from the IgG-inducing antigens by a combination of DEAE-Sephadex A-50 chromatography and Sephadex G-200 gel filtration. The molecular weight of the purified preparation was estimated to be approximately 20,000 by gel filtration. The carbohydrate content of the preparation was apparently low, about 2%. Rats immunized with the allergenic fraction developed only a homocytotropic antibody and no hemagglutination antibody.     

Trichinella spiralis: specificity of ES antigens from pre-encysted larvae.

Excretory/secretory (ES) antigens were obtained by culturing pre-encysted Trichinella spiralis larvae which were recovered from muscles of experimentally infected mice 14-15 days postinfection. Analyses of these antigens (PEL ES) with immunoblotting, SDS-PAGE and Triple Antibody ELISA showed that they yielded a low sensitivity and specificity when tested with antisera against the common nematodes of Chinese pigs. As compared to ES antigens from encysted larvae, PEL ES also contained more low molecular mass proteins.