Use of the Double Antibody and Nitrocellulose Membrane Filtration Techniques to Separate Free Antigen from Antibody Bound Antigen in Radioimmunoassays

Author(s):  
H. VanVunakis ◽  
L. Levine
1979 ◽  
Vol 25 (5) ◽  
pp. 752-756 ◽  
Author(s):  
R K Gupta ◽  
D L Morton

Abstract We compared the protein-A-bearing Staphylococcus aureus immunoadsorbent to the double-antibody precipitation method for separating bound and free radiolabeled antigen in a radioimmunoassay. With human albumin (antigen) and rabbit anti-human albumin (antibody) as a model, our results indicate that formalin-fixed, heat-killed S. aureus cells could be substituted for the double-antibody precipitation method. Ease of preparation and high adsorption capacity of protein-A-bearing S. aureus for most mammalian IgG make this method economical and time saving.


1989 ◽  
Vol 61 (03) ◽  
pp. 386-391 ◽  
Author(s):  
Guido Tans ◽  
Truus Janssen-Claessen ◽  
Jan Rosing

SummaryIn this paper we report a method via which enzymatically active products formed during prothrombin activation can be detected by simple photographic means after SDS-gel electrophoresis, blotting onto a nitrocellulose membrane and visualization with the chromogenic substrate, S2238. After amidolytic detection the same nitrocellulose membrane can also be used for immunologic detection of prothrombin activation products, thus allowing a complete description of product formation during prothrombin activation.The detection limit of the so-called “amidoblot” is approximately 3 ng thrombin per gel sample which is comparable to the sensitivity of immunoblotting.It is further shown that the amidoblot technique can also be applied to other coagulation factors for which a suitable chromogenic substrate is available (factor XIIa, kallikrein, factor XIa, factor Xa, plasmin and activated protein C).


1966 ◽  
Vol 51 (1) ◽  
pp. 88-94 ◽  
Author(s):  
A. Villanueva ◽  
S. J. H. Ashcroft ◽  
J. P. Felber

ABSTRACT The synthetic ACTH peptides β1–39 and β1–24 stimulated lipolysis as determined by the rat epididymal fat pad in vitro. The stimulating effect of these peptides was diminished by prior incubation of the peptides with antibodies produced by the guinea-pig against ACTH. The stimulating effect of these hormones was also diminished by the double antibody system used in the radio-immunoassay of ACTH and other peptide hormones, in which incubation with antiserum is followed by precipitation of the antigen-antibody complex by rabbit anti-guinea-pig-γ-globulin.


1972 ◽  
Vol 71 (1) ◽  
pp. 24-36 ◽  
Author(s):  
Ariel Gordin ◽  
Pirkko Saarinen

ABSTRACT An account is given of a methodological study of the double-antibody radioimmunoassay of human TSH, using highly purified labelled human TSH as tracer. It was shown that conventional paper electrophoresis was not adequate for studying the purity of labelled human TSH. When polyvinylchloride (Pevikon®) electrophoresis was used, four subfractions could still be separated, even though, on paper electrophoresis, the material seemed to be homogeneous. Only two of the four Pevikon fractions were immunoreactive. Purification of labelled human TSH by Pevikon electrophoresis also improved the sensitivity of the assay. Specific activities of about 100 mCi/mg gave the highest initial binding capacity, produced least damage to the labelled hormone and showed the best stability of the tracer without influencing the sensitivity of the method. In different storage conditions, labelled human TSH was found to be most stable at −20°C and diluted 1/100. Only in pregnancy did the addition of HCG seem necessary. The mean TSH value in healthy subjects was 3.6 ± 1.4 μU/ml (mean±sd) with a range from 1.6 μU/ml to 8.8 μU/ml.


2019 ◽  
Vol 56 (6) ◽  
pp. 54-56
Author(s):  
E.Yu. Agarkova ◽  
◽  
A.G. Kruchinin ◽  
A.A. Agarkov ◽  
V.D. Haritonov

2010 ◽  
Vol 5 (4) ◽  
Author(s):  
J. L. Manuszak ◽  
M. MacPhee ◽  
S. Liskovich ◽  
L. Feldsher

The City of Baltimore, Maryland is one of many US cities faced with challenges related to increasing potable water demands, diminishing fresh water supplies, and aging infrastructure. To address these challenges, the City recently undertook a $7M study to evaluate water supply and treatment alternatives and develop the conceptual design for a new 120 million gallon per day (MGD) water treatment plant. As part of this study, an innovative raw water management tool was constructed to help model source water availability and predicted water quality based on integration of a new and more challenging surface water supply. A rigorous decision-making approach was then used to screen and select appropriate treatment processes. Short-listed treatment strategies were demonstrated through a year-long pilot study, and process design criteria were collected in order to assess capital and operational costs for the full-scale plant. Ultimately the City chose a treatment scheme that includes low-pressure membrane filtration and post-filter GAC adsorption, allowing for consistent finished water quality irrespective of which raw water supply is being used. The conceptual design includes several progressive concepts, which will: 1) alleviate treatment limitations at the City's existing plants by providing additional pre-clarification facilities at the new plant; and 2) take advantage of site conditions to design and operate the submerged membrane system by gravity-induced siphon, saving the City significant capital and operations and maintenance (O&M) costs. Once completed, the new Fullerton Water Filtration Plant (WFP) will be the largest low-pressure membrane plant in North America, and the largest gravity-siphon design in the world.


2003 ◽  
Vol 3 (3) ◽  
pp. 43-48
Author(s):  
J.Y. Huang ◽  
T. Uchiburi ◽  
K. Fujita

Electrolytic coagulation has advantages compared with conventional chemicals coagulation treatment. First of all, the addition of alkaline agent is necessary in chemicals coagulation treatment as the addition of acid agents in coagulation of dense wash water makes pH decline. While not much change of pH is observed with electrolytic coagulation, where alkaline agent is not demanded. Secondly, it is easy to control the dose of additional coagulants. In this study, electrolytic coagulation sedimentation treatment is adopted for membrane retentate from membrane filtration treatment without using coagulants. As electricity can control the addition of coagulation, an automation method is applied in the experiment. The lower the iron as well as aluminum dissolved, the higher the sludge density becomes; especially aluminum has this tendency. It is possible to achieve 400 kg/m3 of condensed sludge density with artificial wash water and more than 80 kg/m3 for real retentate, in terms of electrolytic coagulation of iron anode. The sludge density of aluminum anode is higher, but its filtrate density is lower than iron anode.


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