sepharose and Hyperlipoproteinemia-Type-II

Topics: Blood Component Removal; Cholesterol; Dextran Sulfate; Equipment Design; History, 20th Century; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hyperlipoproteinemia Type II; Lipoproteins; Plasma Exchange; Sepharose

Topics: Animals; Arteriosclerosis; Blood Component Removal; Dextran Sulfate; Extracorporeal Circulation; Heparin; Humans; Hyperlipoproteinemia Type II; Lipoproteins, LDL; Plasmapheresis; Sepharose; Sorption Detoxification

Immunoadsorption is an application of affinity chromatography, as a therapeutic method to specifically deplete biological fluids such as blood plasma from proteins in excess, or to extract a biomolecule from a complex mixture. However, the leakage of small amounts of antibodies covalently immobilized on the support hampers the practical use of this method. In fact, these released antibodies contaminate the purified proteins or depleted media and, when they are of animal nature, they may lead to immunization of patients, or cause an anaphylactic shock when a clinical use is concerned. It is therefore of prime importance that the immunoadsorbents exhibit a satisfactory stability over the whole range of chemical and biochemical conditions involved during their clinical handling. To determine optimal conditions for the preparation of stable immunoadsorbents designed to remove selectively Low Density Lipoproteins (LDLs) from the plasma of patients affected by familial hypercholesterolemia, various immunoadsorbents were prepared by covalent immobilization of goat anti-apolipoprotein B polyclonal antibodies on different supports (Sepharose CL-4B, Sepharose 6 Fast Flow, Sphérodex and Fractogel) previously activated by various chemical reagents (cyanogen bromide, divinyl sulphone, tresyl chloride and trichloro-s-triazine). Their adsorption capacity, specificity, stability and the amount of immobilized antibodies were compared in terms of the activation method and the support used. It turns out that the immunoadsorbents prepared with Sepharose 6 Fast Flow lead to optimal yield of coupling, adsorption capacity, and an excellent stability at neutral pH. TC-activated-Fractogel turns out as well to afford an excellent coupling yield, a good adsorption capacity and an optimal stability in the whole pH range tested.

Topics: Adsorption; Antibodies; Apolipoproteins B; Chromatography, Affinity; Cyanogen Bromide; Extracorporeal Circulation; Humans; Hyperlipoproteinemia Type II; Immunosorbents; Sepharose

Low levels of high-density lipoproteins (HDLs) may constitute an independent risk factor that may be as important as elevated low-density lipoproteins (LDLs) in coronary artery disease (CAD). Concentrations and distributions of lipids, apolipoprotein (apo) B, and apoA-I in the plasma and lipoprotein subfractions of two groups of swine, one with familial hypercholesterolemia (FHC) and the other with diet-induced hypercholesterolemia (DHC), were examined. Normolipidemic (NL) animals served as controls. All pigs carried the Lpb5 apoB mutation, which is known to influence the formation of atherosclerotic lesions. Mean concentrations of serum total cholesterol in NL, DHC, and FHC were 80.0 +/- 9.3, 774.3 +/- 54.5, and 316.5 +/- 36.1 mg/dL, respectively; HDL cholesterol (HDL-C), 33.5 +/- 1.9, 137.0 +/- 9.9, and 22.3 +/- 2.2 mg/dL; triglycerides, 33.0 +/- 16.3, 40.3 +/- 11.7, and 56.8 +/- 7.2 mg/dL; apoB, 35.7 +/- 3.1, 142.0 +/- 4.8, and 169.3 +/- 13.9 mg/dL; and apoA-I, 62.4 +/- 9.3, 170.9 +/- 6.9, and 42.6 +/- 4.8 mg/dL. The distributions of total cholesterol, apoB, and apoA-I in plasma lipoprotein subfractions were also examined. Compared with NL, FHC had fourfold and 4.7-fold increases in total cholesterol and apoB, respectively, distributed in the lower densities (d < 1.043 g/mL), and low HDL-C and apoA-I levels, resulting in a high total cholesterol/HDL-C ratio (14.4:1) and elevated triglyceride levels. DHC was characterized by 10-fold and fourfold increases in total cholesterol and apoB, respectively, resulting in an LDL particle highly enriched in cholesterol, a fourfold increase of HDL-C, an almost threefold increase in apoA-I, and a normal triglyceride level.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Apolipoprotein A-I; Apolipoproteins B; Cholesterol; Diet; Disease Models, Animal; Electrophoresis, Polyacrylamide Gel; Gels; Hypercholesterolemia; Hyperlipoproteinemia Type II; Immunoelectrophoresis; Lipids; Lipoproteins; Male; Sepharose; Sodium Dodecyl Sulfate; Swine

In summary the therapeutic use of affinity chromatography, the immunapheresis with LDL-Therasorb columns, Baxter, has been shown to be a safe, specific and efficacious extracorporeal treatment. Following GMP rules LDL-Therasorb is produced in a constant quality. Beyond the treatment of hypercholesterolemia the versatility of antibodies opens this technology to the treatment of other diseases.

Topics: Antibody Specificity; Blood Component Removal; Cholesterol; Chromatography, Affinity; Humans; Hyperlipoproteinemia Type II; Immunosorbent Techniques; Lipoproteins, LDL; Risk Factors; Sepharose

Heparin-sorbent was developed in the USSR Cardiology Research Center for LDL-apheresis in patients with familial hypercholesterolemia. Clinical trials of the sorbent were started in 1985. Heparin-sorbent has a high capacity for LDL. One column (400ml) can remove 3 - 6g of LDL cholesterol, depending on the initial LDL content in the plasma. The sorbent demonstrates a high stability, it can withstand more then 75 sorbtion-desorbtion cycles with only a slight loss of the initial capacity. It means that one column can work for 1,5-2,0 years when used weekly. Heparin-sorbent is the sorbent of limited selectivity, apo-B content in eluate is 40-60% of the total protein content, but it is important that it does not bind HDL.

Topics: Binding Sites; Chromatography, Affinity; Heparin; Humans; Hyperlipoproteinemia Type II; Lipoproteins, LDL; Plasmapheresis; Sepharose; Sodium Chloride

Plasmosorption on heparin sepharose columns was shown to cause a decrease in the concentration of atherogenic lipoproteins. The results obtained suggest that plasmosorption results in the improvement of the patients' status, a decrease in the frequency and intensity of anginal attacks. Columns can be effectively used after repeated regenerations up to 30-40 times.

Topics: Adult; Angina Pectoris; Cholesterol; Coronary Artery Disease; Female; Hemoperfusion; Heparin; Humans; Hyperlipoproteinemia Type II; Lipoproteins, LDL; Male; Sepharose; Time Factors

A method is described for the removal of cholesterol fractions by means of intermittent extracorporeal plasma circulation through a column containing heparin-treated agarose in the presence of calcium. The system allows plasma perfusion in one column while simultaneously recharging another. In vitro testing demonstrated its efficiency, and in vivo testing of a normal dog proved its feasibility. In vitro testing revealed a mean removal of 22% of low-density and very low-density lipoproteins combined. Animal testing did not produce any abnormalities in clinically significant parameters.

Topics: Animals; Cholesterol; Dogs; Hemoperfusion; Heparin; Humans; Hyperlipoproteinemia Type II; In Vitro Techniques; Lipoproteins; Plasmapheresis; Sepharose

Extracorporeal removal of cholesterol from blood with heparin-agarose conjugates was studied as a means of reducing the plasma cholesterol concentration in two patients with homozygous familial hypercholesterolemia. Both patients were treated in an outpatient clinic. Patient 1 underwent two separate courses of treatment; during the first course of eight treatments, the plasma cholesterol concentration decreased by a maximum of 54% (from 811-370 mg/dl). The six treatments in the second course resulted in a 35% reduction of plasma cholesterol (from 939--632 mg/dl). In patient two, 12 consecutive treatments resulted in a 56% decrease in the concentration of plasma cholesterol (from 768--298 mg/dl). Repetitive treatments in both patients resulted in a new "steady state" with circulating cholesterol levels lower than the pretreatment baseline value. The decrease in plasma cholesterol is mainly due to the removal of low density lipoproteins. The circulating level of high density lipoproteins is unaffected by the treatment. The treatment has no effect on the chemical composition of both low and high density lipoproteins. The extracorporeal treatment of blood with heparin-agarose was well tolerated by both patients and there were no undesirable effects. The effectiveness and simplicity of the present approach makes it attractive as a possible mode of therapy for homozygous familial hypercholesterolemia.

Topics: Adolescent; Child, Preschool; Evaluation Studies as Topic; Hemoperfusion; Heparin; Humans; Hyperlipoproteinemia Type II; Lipoproteins, LDL; Male; Polysaccharides; Sepharose