Binding of C1q and DNA to support materials by means of a new coupling procedure.

Since several years matrix-bound biologically active substances are widely used in biosciences, biotechnology and medicine. We are presenting a simple and inexpensive activation procedure for support materials which allows stable binding of C1q and DNA. We could not found a remarkable leakage of the bound ligands. In connection with the good biocompatibility of our procedure, both are important for the application in extracorporeal immunoadsorption. The method is technologically simple and represents an alternative to the activation methods applied so far.

Determination of acetylcholine and choline by flow-injection with immobilized enzymes and fluorometric or luminometric detection.

A method for determination of picomolar quantities of acetylcholine and choline in solutions and tissue extracts is described. The analytes are injected into a continuous stream of a simple medium flowing through a sequence of enzyme reactors containing acetylcholinesterase, choline oxidase, and peroxidase. Additional reactors with choline oxidase and catalase are used to remove endogenous choline from the tissue extracts in which the content of acetylcholine is to be measured. Reaction products are detected fluorometrically or luminometrically. The limits of sensitivity are about 10 pmol/sample with luminometric and 0.2 pmol/sample with fluorometric detection.

Influence of salts on the covalent immobilization of proteins to modified copolymers of 2-hydroxyethyl methacrylate with ethylene dimethacrylate.

In the study of the covalent immobilization of aminoacylase, thermitase, pepsin, trypsin, chymotrypsin, elastase, subtilisin, penicillinamidohydrolase, carboxypeptidase A, cystathionine-beta-synthase, and anticathepsin D-IgG to copolymers of 2-hydroxyethyl methacrylate and ethylene dimethacrylate (Separon HEMA) containing epoxy groups a marked influence of added salts on the immobilization efficiency was observed. Yields in covalently bound active enzymes were dependent on the concentrations and type of ions added, which can be arranged according to the Hofmeister series. At a distinct concentration, the salting-out ions cause a protein-matrix hydrophobic interaction which is a prerequisite for the covalent bond formation.

The mitogenicity of extracts from Sarcocystis gigantea macrocysts is due to lectin(s).

As recently reported, extracts from macrocysts of Sarcocystis gigantea obtained by sonication show mitogenic activity towards lymphocytes. By affinity chromatography using a galactose gel, a lectin fraction which contained the entire mitogenic activity was obtained from these extracts.

Separation of human lymphoid cells by affinity chromatography and cell surface labelling by hydroxyethyl methacrylate particles using monoclonal antibodies.

The hydroxyalkyl methacrylate gel Separon, Sepharose 6MB and polystyrene dishes with bound antibodies were used for affinity chromatography of cells isolated from human tonsils and peripheral blood. Enrichment of T cells by filtration through anti-human immunoglobulin Separon or Sepharose 6MB columns was comparable to that achieved by "panning" on dishes or by Nylon wool column fractionation. High enrichment of B cells was obtained when cells pretreated with monoclonal anti-T cell antibodies were fractionated on columns or dishes where anti-mouse antibody was bound. Hydroxyethyl methacrylate copolymer microspheres were conjugated with monoclonal antibodies directed against surface markers of human lymphocytes or with anti-mouse immunoglobulin antibodies and used for surface labelling of cells. The results obtained by immunofluorescence were in good correlation with labelling by hydroxyethyl methacrylate immunobeads.

Ion-exchange derivatives of Spheron. V. Sulphate and sulpho derivatives.

Using various reactions, strongly acidic derivatives of Spheron of the sulphate and sulpho types were prepared with nominal capacities between 0.37 and 2.0 mequiv./g. They were characterized by their capacities for small ions, by their static and dynamic capacities for proteins, by elemental analysis, by their working volumes and specific internal surface areas. The differences between the nominal capacities calculated theoretically from the sulphur contents and the measured values are discussed, as are the relationships between the nominal capacities for small ions and the static and dynamic capacities for proteins. The chromatographic properties of the cation exchangers are illustrated by the separation of an artificial mixture of proteins (serum albumin, chymotrypsinogen and lysozyme) at various pH values, and of a natural mixture of egg-white proteins. The separation of active enzymes is also reported. Sulphobutyl-Spheron was tested also as a catalyst for the esterification of alcohols.

The significance of vaginal hormonal cytology and the leukocyte adherence inhibition test for the detection of malignancy.

Raised leukocyte adherence inhibition (LAI) test values were found for 25 women with endometrial carcinoma and 12 women with benign hyperplastic endometrium. Of 42 patients with oligomenorrheic and polymenorrheic disturbances of the menstrual cycle and whose hyperestrogenism was assessed by cytologic indices (i.e., eosinophilic and karyopyknotic indices), 38 exhibited positive or borderline LAI test values with either one or both of the antigens used. The LAI test was negative with both antigens in four women with hyperestrogenism associated with a 28-day menstrual cycle. The high correlation between hyperestrogenism and positive LAI test results as well as between the LAI test and the presence of tumors suggests that both hormonal cytology and the LAI test might be useful as screening methods for evaluating risk factors for the development of malignant tumors of the female reproductive tract. Both methods are fairly simple, reproducible and convenient for long-term follow-up.