Complement C3 in serum and plasma, as measured by radial immunodiffusion with four commercial kits.

1976 ◽  
Vol 22 (2) ◽  
pp. 267-269 ◽  
Author(s):  
A O Vladutiu ◽  
B M Winiarski
Keyword(s):  
Radial Immunodiffusion ◽  
Complement C3 ◽  
Antigenic Determinants ◽  
Sample Processing ◽  
Single Radial Immunodiffusion ◽  
Normal Individuals ◽  
Commercial Kits

Abstract Quantitation of the C3 component of complement by single radial immunodiffusion is subject to error because the C3 molecule has several antigenic determinants and anti-C3 sera differ in their specificity to these determinants. C3 was measured in plasma (ethylenediaminetetraacetate anticoagulant) and serum, in five normal individuals, by use of four such commercial kits. The effects of ethylenediaminetetraacetate and of incubation at -20, 4, and 37 degrees C for 1, 2, and 7 days were investigated. We found wide variations in C3 in the same sample, as measured with different kits. Incubation for longer than 48 h at 4 degrees C changed C3 concentrations as compared to those in fresh samples. Reference sera for C3 assay and standardized procedures for sample processing are both essential to valid results.

Immunochemical Measurement of Lipoprotein-X

1974 ◽  
Vol 20 (6) ◽  
pp. 676-681 ◽  
Author(s):  
Gerhard M Kostner ◽  
Walter Petek ◽  
Anton Holasek
Keyword(s):  
Coefficient Of Variation ◽  
Peak Height ◽  
Normal Serum ◽  
Radial Immunodiffusion ◽  
Antigenic Determinants ◽  
Globulin Fraction ◽  
Single Radial Immunodiffusion ◽  
Immunochemical Methods

Abstract Lipoprotein-X, an abnormal lipoprotein that is specific for cholestasis, was quantitated by immunochemical methods. Interfering lipoproteins also present in normal serum and sharing antigenic determinants with lipoprotein-X were removed before the sample was applied, by precipitation with purified anti-lipoprotein B or a γ-globulin fraction of specific lipoprotein B antiserum. On Laurell electrophoresis, peak height was linearly related to lipoprotein-X concentration in the range 0.20-10 g/liter of serum. Sensitivity could be increased further by staining the plates. The coefficient of variation was less than 5%. Single radial immunodiffusion (Mancini et al. technique) was somewhat less sensitive and accurate. Results were available after 3 h by Laurell's electroimmunodiffusion technique, and after 72 h by the technique of Mancini et al. Equivalent results were obtained for samples of lipoprotein-X of extrahepatic or intrahepatic origin.

Single radial immunodiffusion of serum apolipoproteins C-II, C-III and E — pretreatment of samples with surfactant

1986 ◽  
Vol 161 (3) ◽  
pp. 275-282 ◽  
Author(s):  
Koji Itakura ◽  
Takeyasu Matsudate ◽  
Takato Sakurai ◽  
Shunichi Hashimoto ◽  
Kazue Ito ◽  
...  
Keyword(s):  
Radial Immunodiffusion ◽  
Single Radial Immunodiffusion

IDENTIFICATION OF ANTIPLATELET ANTIBODY IDIOTYPlSS ASSOCIATED WITH GLYCOPROTEIN Ib SPECIFICITY, PRESENT IN ITP PLASMA AND PRODUCED BY HUMAN HYBRIDOMAS FROM ITP SPLEEN CELL FUSIONS

1987 ◽  
Author(s):  
Diane Nugent
Keyword(s):  
Severe Form ◽  
Antigenic Determinants ◽  
Membrane Glycoproteins ◽  
Cellular Regulation ◽  
Antiplatelet Antibody ◽  
Autoantibody Production ◽  
Normal Individuals ◽  
Clonal Origin

Platelet membrane glycoproteins (GP) express anumber of antigenic determinants important in theetiology of autoimmune thrombocytopenia. Sensitization to GPIb, although not the most frequent cause of ITP, leads to a particularly severe form ofthe disease. We have identified a number of casesof ITP in which GPIb bears the relevant immunogen. Using GPIb-specific autoantibodies isolated from the plasma of one such patient, we have produced a number of rabbit polyclonal and murine monoclonal anti-idiotypic antibodies. These antibodies recognize an idiotype expressed on the IgM antibody of this patient as well as IgG or IgM antibodies from several other patients with ITP, all of which can be shown to bind specifically to GPIb. Statistical analysis of a series of plasmas from normal individuals and thrombocytopenic patients demonstrated that there is a very strong correlation between the presence of the idiotype and GPIb reactivity, (p < 0.00001). These anti-idiotypic antibodies are useful for the detection and characterization of GPIb-specific antibodies in the sera of patients with a clinically severe form of ITP. The classification of patients bearing this idiotype in their plasma may be useful in predicting disease outcome, thus identifying a group of ITP patients in whom more aggressive therapeutic regimens may be indicated. The use of these reagents and the development of human B lymphoblastoid cell lines producing monoclonal anti-GPIb antibodies will serve to elucidate the clonal origin and cellular regulation of autoantibody production in this disease

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