Gel Entrapment of Antibody: A New Strategy for Facilitating Both Manual and Automated Radioimmunoassay

1973 ◽  
Vol 19 (12) ◽  
pp. 1339-1344 ◽  
Author(s):  
Stuart J Updike ◽  
John D Simmons ◽  
Douglas H Grant ◽  
Judith A Magnuson ◽  
Theodore L Goodfriend
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Room Temperature ◽  
Solid Phase ◽  
Binding Activity ◽  
Angiotensin I ◽  
Chromatographic Columns ◽  
New Strategy ◽  
Sample Application ◽  
Solid Phase Binding

Abstract A technique of radioimmunoassay is presented that eliminates pipetting and centrifugation, and excludes interferences by high-molecular-weight materials from the incubation and separation steps. A solid-phase binding reagent is prepared by first entrapping antibody in polyacrylamide gel. This gel is then fragmented, sieved, dried with ethanol or lyophilized, and placed in miniature disposable chromatographic columns. Application of the sample to the intra-gel column compartment is determined by the water regain of the gel. This pipetless method of sample application depends on reproducible aliquots of dry gel particles in every column. A method for preloading radiolabeled hormone and standard hormone into the column is also described. This technique has been successfully applied to the assay of angiotensin I and insulin. Dry antibody—gel stored at room temperature for 26 months has not shown loss of binding activity.

Comparative analysis of the transferrin and lactoferrin binding proteins in the family Neisseriaceae

1989 ◽  
Vol 35 (3) ◽  
pp. 409-415 ◽  
Author(s):  
Anthony B. Schryvers ◽  
B. Craig Lee
Keyword(s):  
Binding Proteins ◽  
Solid Phase ◽  
Bacterial Species ◽  
Binding Activity ◽  
Intact Cells ◽  
Human Transferrin ◽  
Affinity Isolation ◽  
Lactoferrin Receptor ◽  
Solid Phase Binding ◽  
Branhamella Catarrhalis

Intact cells of several bacterial species were tested for their ability to bind human transferrin and lactoferrin by a solid-phase binding assay using horseradish peroxidase conjugated transferrin and lactoferrin. The ability to bind lactoferrin was detected in all isolates of Neisseria and Branhamella catarrhalis but not in isolates of Escherichia coli or Pseudomonas aeruginosa. Transferrin-binding activity was similarly detected in most isolates of Neisseria and Branhamella but not in E. coli or P. aeruginosa. The expression of transferrin- and lactoferrin-binding activity was induced by addition of ethylenediamine di-o-phenylacetic acid and reversed by excess FeCl3, indicating regulation by the level of available iron in the medium. The transferrin receptor was specific for human transferrin and the lactoferrin receptor had a high degree of specificity for human lactoferrin in all species tested. The transferrin- and lactoferrin-binding proteins were identified after affinity isolation using biotinylated human transferrin or lactoferrin and streptavidin–agarose. The lactoferrin-binding protein was identified as a 105-kilodalton protein in all species tested. Affinity isolation with biotinylated transferrin yielded two or more proteins in all species tested. A high molecular mass protein was observed in all isolates, and was of similar size (approximately 98 kilodaltons) in all species of Neisseria but was larger (105 kilodaltons) in B. catarrhalis.Key words: iron, Neisseria, transferrin, lactoferrin, receptor.

An Immunoprecipitation Assay for High Molecular Weight Alkaline Phosphatase in Human Serum

1989 ◽  
Vol 26 (2) ◽  
pp. 151-157 ◽  
Author(s):  
Gerald A Maguire ◽  
Halima Adnan
Keyword(s):  
Molecular Weight ◽  
Alkaline Phosphatase ◽  
Liver Disease ◽  
High Molecular Weight ◽  
Solid Phase ◽  
Bone Alkaline Phosphatase ◽  
Intestinal Alkaline Phosphatase ◽  
Alkaline Phosphatases ◽  
Molecular Weight Form ◽  
Hepatic Malignancies

The serum of patients with obstructive liver disease may contain a high molecular weight form of alkaline phosphatase (high Mr alkaline phosphatase). The presence of this form of alkaline phosphatase is associated with hepatic malignancies. We have investigated the use of anti-alkaline phosphatase monoclonal antibodies which do not bind high Mr alkaline phosphatase in assays for high Mr alkaline phosphatase. Direct immunoprecipitation of liver and bone alkaline phosphatase with solid phase anti-liver alkaline phosphatase antibody (which also reacts with bone alkaline phosphatase) and measurement of the residual supernatant alkaline phosphatase activity led to a precise assay. Intestinal alkaline phosphatase interfered in this assay which, consequently, was of little use in the differential diagnosis of liver disease. Indirect precipitation of liver, bone, placental and intestinal alkaline phosphatase by soluble anti-liver alkaline phosphatase (which reacts with liver and bone alkaline phosphatases), soluble anti-intestinal alkaline phosphatase (which reacts with placental and intestinal alkaline phosphatases) and solid phase anti-mouse IgG led to an assay which, although less precise, showed more promise of being useful clinically.

Intraindividual Comparison of the Impact of two Selective Apheresis Methods (DALI and HELP) on the Coagulation System

2000 ◽  
Vol 23 (3) ◽  
pp. 199-206 ◽  
Author(s):  
U. Julius ◽  
G. Siegert ◽  
S. Gromeier
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Protein S ◽  
Binding Activity ◽  
Coagulation System ◽  
Factor Xii ◽  
High Molecular Weight Kininogen ◽  
Prothrombin Fragment ◽  
Intraindividual Comparison ◽  
The Impact

We performed an intraindividual comparison of the effect on the coagulation system of two selective apheresis procedures: Direct Adsorption of Lipoproteins (DALI) and Heparin-induced Lipoprotein Fibrinogen Precipitation (HELP). Six patients suffering from heterozygous familial hypercholesterolemia have been treated with 2 sessions of each procedure. Anticoagulation was carried out according to usual recommendations. Blood samples were taken before, immediately after and on the second day after the sessions. We assessed global coagulation tests (prothrombin time, activated partial thromboplastin time), fibrinogen, prothrombin fragment F 1 + 2 and a variety of factors (Factors II, V, VII, XIII, IX, X, XI, XII, XIIa; von Willebrand Factor; collagen-binding activity, prekallikrein, high-molecular weight kininogen) and antagonists (antithrombin III, protein S activity, free protein S). In fact, all parameters measured have been influenced by the apheresis treatment. Fibrinogen is lowered more by HELP, which also has a more definite impact on factors belonging to the prothrombin complex (II, VII, X). In contrast, the major effects of the DALI system have been seen on the intrinsic pathway of the coagulation system (IX, XI, prekallikrein, high-molecular-weight kininogen). With both systems, no increases in activated Factor XII or in prothrombin fragment F1 + 2 have been observed. These data provide a solid basis for individual adaptations of anticoagulant doses.

Solid-phase polycondensation for producing polyethylene terephthalate with a high molecular weight

1974 ◽  
Vol 5 (5) ◽  
pp. 493-495
Author(s):  
M. I. Simonova ◽  
E. M. Aizenshtein ◽  
V. V. Shevchenko
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Polyethylene Terephthalate ◽  
Solid Phase

Selective or living organopolymerization of a six-five bicyclic lactone to produce fully recyclable polyesters

2019 ◽  
Vol 10 (23) ◽  
pp. 3097-3106 ◽  
Author(s):  
Robin M. Cywar ◽  
Jian-Bo Zhu ◽  
Eugene Y.-X. Chen
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Room Temperature ◽  
Heterocyclic Carbenes ◽  
Base Pairs ◽  
Ring Open ◽  
Bicyclic Lactone

A ring-fused γ-butyrolactone can be selectively ring-open polymerized at room temperature by N-heterocyclic carbenes to cyclic polyester or by bifunctional (thio)urea and base pairs in a living fashion to high molecular weight linear polyester that can be organocatalytically and quantitatively recycled at 120 °C.

Formation of high-molecular weight polyaminoborane by Fe hydride catalysed dehydrocoupling of methylamine borane

2017 ◽  
Vol 46 (21) ◽  
pp. 6843-6847 ◽  
Author(s):  
F. Anke ◽  
D. Han ◽  
M. Klahn ◽  
A. Spannenberg ◽  
T. Beweries
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Room Temperature ◽  
Catalyst Precursor

The complex [(PNHP)Fe(H)(CO)(HBH3)] (PNHP = HN(CH2CH2Pi-Pr2)2) serves as a catalyst precursor for the selective dehydrocoupling of methylamine borane at room temperature, tentatively via an off-metal polymerisation pathway.

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