Study of parameters involved in specific immunoadsorption of apolipoprotein B.

A therapeutic immunoadsorption system on immobilized and anti-apolipoprotein B as a plasma cholesterol lowering procedure was optimized. Several antibodies were compared and highest adsorption capacity was obtained with goat polyclonal antibodies. Optimum quantities of antibodies to be immobilized on the gel and quantities of apo-B to be applied to columns were determined. The amount of antibodies released from immunoadsorbents can be minimized by treatment with a 0.005% glutaraldehyde solution with an acceptable reduction rate of adsorption capacity. Each phase, adsorption and desorption respectively, were well-defined and synchronized so two columns could be used in parallel in an automated procedure. In these conditions, the immunoadsorption system can efficiently, specifically and safely remove cholesterol and has to be subjected to clinical trials.

[Development of a technique of immunoadsorption of LDL-cholesterol]. / Mise au point d'une technique d'immuno-adsorption du LDL-cholestérol.

An immunoadsorption system for lowering plasma cholesterol was optimized. Several polyclonal and monoclonal antibodies were compared and the best results were obtained with goat polyclonal antibodies. The optimum quantity of antibodies to be immobilized on the gel was 5 mg/ml. Taking into account two variables, i.e., 1) that the regeneration must be as complete as possible and, 2) that immunoadsorbents must be used several times without a loss of adsorption capacity, desorption was achieved with 0.3 M glycine adjusted to pH 2.8. Antibody release from the immunoadsorbent was determined and can be minimized by glutaraldehyde treatment of the immunoadsorbent. Each phase, adsorption and desorption, respectively, was well-defined and synchronized, so that two columns could be used in parallel in an automated procedure. The kinetics of plasma protein removal demonstrated the efficiency and the specificity of the procedure.

[Immunosorption of apolipoproteins B. Optimization of the method in an in vitro bench test]. / Immunoépuration des apolipoprotéines B. Optimisation de la méthode sur banc d'essai in vitro.

The aim of the present study was to determine optimum conditions for preparation and use of immunoadsorbents for removal of apolipoprotein B from plasma. An in vitro study was carried out to optimize different chromatography parameters: linear flow rate, apo B quantity loaded, gel regeneration conditions. Specificity of removal of apo B, stability of antibody covalent linkage and possible multiple use of immunoadsorbents were assessed. No significant loss of adsorption capacity was observed after sterilization and storage of immunoadsorbents.