scholarly journals A plasma protein fractionation procedure for use in studies of protein metabolism. Its application to the measurement of synthesis rates of two α1-glycoproteins and other serum proteins in the rat

1974 ◽  
Vol 141 (3) ◽  
pp. 655-665 ◽  
Author(s):  
J. Ho ◽  
K. N. Jeejeebhoy ◽  
R. H. Painter
Keyword(s):  
Electrophoretic Mobility ◽  
Plasma Protein ◽  
Protein Metabolism ◽  
Plasma Proteins ◽  
Serum Proteins ◽  
Single Sample ◽  
Protein Fractionation ◽  
Step Method ◽  
Preparative Scale ◽  
Fractionation Procedure

A two-step method for the separation of five different plasma proteins on a preparative scale, which is capable of being extended to allow the separation of other plasma proteins, is described. The proteins separated were fibrinogen, two α1-glycoproteins, albumin and transferrin. The α1-glycoproteins were characterized in terms of electrophoretic mobility, ultracentrifugal and immunological characteristics. By using this method, it was shown that a single sample of plasma could be fractionated to yield purified proteins in sufficient quantity to simultaneously measure the synthesis of the two α1-glycoproteins, albumin and transferrin in the rat with McFarlane's technique (McFarlane, 1963; Reeve et al., 1963; McFarlane et al., 1965).

Evaluation of electrophoretic mobility shift assay as a method to determine plasma protein binding of siRNA

Bioanalysis ◽  
2019 ◽  
Vol 11 (21) ◽  
pp. 1927-1939 ◽  
Author(s):  
Carrie Rocca ◽  
Sean Dennin ◽  
Yongli Gu ◽  
Joohwan Kim ◽  
Samantha Chigas ◽  
...  
Keyword(s):  
Protein Binding ◽  
Electrophoretic Mobility Shift Assay ◽  
Electrophoretic Mobility ◽  
Plasma Protein Binding ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Small Interfering Rnas ◽  
Mobility Shift ◽  
Mobility Shift Assay

Aim: The electrophoretic mobility shift assay (EMSA) was evaluated as an alternative to ultrafiltration (UF) to assess plasma protein binding (PPB) of small interfering RNAs (siRNA) and antisense oligonucleotides (ASO). Results & methodology: EMSA analysis showed that PPB depended on siRNA and plasma concentration. Conversely, when analyzed by ultrafiltration, siRNA bound the filtration device nonspecifically and PPB remained >98% across physiologically relevant siRNA concentrations. Using EMSA, siRNA exhibited charge-based interactions with plasma proteins, while ASO remained highly bound to plasma proteins or albumin in the presence of 500 mM salt. Conclusion: PPB characteristics of siRNA and ASO can be distinguished using EMSA. Characterization of siRNA PPB by EMSA enhances our knowledge of siRNA absorption, distribution, metabolism and excretion and advanced development of RNA interference therapeutics.

scholarly journals PLASMA PROTEIN METABOLISM—NORMAL AND ASSOCIATED WITH SHOCK

1944 ◽  
Vol 80 (6) ◽  
pp. 455-475 ◽  
Author(s):  
R. M. Fink ◽  
T. Enns ◽  
C. P. Kimball ◽  
H. E. Silberstein ◽  
W. F. Bale ◽  
...  
Keyword(s):  
Plasma Protein ◽  
Protein Metabolism ◽  
Plasma Proteins ◽  
Blood Stream ◽  
Dilution Curve ◽  
Body Protein ◽  
Protein Mass ◽  
Rapid Exchange ◽  
Isotope Concentration ◽  
Heavy Nitrogen

Labeled plasma proteins are produced by administering to dogs the amino acid lysine synthesized with heavy nitrogen. Such labeled proteins are apparently indistinguishable biologically from proteins of normal isotope concentration. Labeled plasma proteins, as plasma, injected into normal dogs pass out of the blood stream at an initially rapid but constantly decreasing non-logarithmic rate. This outflow is balanced by a simultaneous inflow of plasma proteins from the tissues. Fifty per cent of the labeled protein is out of the blood stream in about 24 hours; 75 per cent in about 6 days. Shock due to trauma of intestine or leg shows a dilution curve of labeled plasma protein not unlike that of the normal dog. If anything, dilution appears a little less rapid in shock. Since the usual shrinkage of plasma volume and plasma protein mass is present in these shocked dogs, these data are compatible with a decreased inflow of protein into the plasma during shock. Methods are described which are suitable for the use of heavy nitrogen incorporated in the epsilon group of lysine and its subsequent analysis in body fluids. These data may indicate that the plasma proteins are normally in constant and rapid exchange with a mobile pool of body protein.

scholarly journals EFFECTS OF LARGE INFUSIONS OF HETEROLOGOUS SERUM PROTEINS ON THE SERUM PROTEIN METABOLISM OF RABBITS

1955 ◽  
Vol 101 (3) ◽  
pp. 233-244 ◽  
Author(s):  
Frank J. Dixon ◽  
Paul H. Maurer
Keyword(s):  
Protein Synthesis ◽  
Serum Protein ◽  
Plasma Protein ◽  
Protein Metabolism ◽  
Serum Proteins ◽  
Foreign Protein ◽  
Protein Catabolism ◽  
Homologous Proteins ◽  
Protein Pool ◽  
Heterologous Serum

Infusions of heterologous serum proteins large enough to replace the entire normal catabolic loss of the corresponding autologous proteins in the recipient rabbits caused increased rates of plasma protein catabolism, an increase in the size of the plasma protein pool and normal or even slightly increased rates of plasma protein synthesis. The principal proteins in these infusions were catabolized at rates similar to those for corresponding homologous proteins. The most marked hyperproteinemias which developed were caused principally by increases in the host's own globulin and to a lesser extent by the presence of foreign protein in the circulation.

Modified Crossed Immunoelectrophoresis to Study with Whole Plasma the Reversible Complex Formation of Histidine-Rich Glycoprotein with Plasminogen

1988 ◽  
Vol 60 (03) ◽  
pp. 411-414 ◽  
Author(s):  
C Kluft ◽  
P Los
Keyword(s):  
Complex Formation ◽  
Electrophoretic Mobility ◽  
Plasma Protein ◽  
Plasma Proteins ◽  
Molecular Forms ◽  
Agarose Gel ◽  
Modified Technique ◽  
Crossed Immunoelectrophoresis ◽  
Dose Dependent

SummaryTo study the reversible complex formation between the plasma protein histidine-rich glycoprotein (HRG) and plasminogen, crossed immunoelectrophoresis of HRG was modified. In the modification, purified plasminogen was introduced into the gel of the first dimension electrophoresis.Two molecular forms of plasminogen, Glu- and Lys-plasmino-gen, induced a dose-dependent reduction of the electrophoretic mobility of HRG, with a half maximal retardation for both plasminogens at 0.50–0.55 μM of added plasminogen to the agarose gel. HRG in plasma behaved as a uniform fraction with respect to plasminogen binding. In contrast, with the same modified technique another plasma protein, β2-antiplasmin, separated into a retarded plasminogen-binding form and a nonretarded non-plasminogen-binding form.The method can be used to assess several aspects of reversible complex formation between plasma proteins, as demonstrated for plasminogen binding of HRG and α2-antiplasmin in whole plasma.

Effects of training and competitive swimming on serum proteins

1961 ◽  
Vol 16 (5) ◽  
pp. 807-809 ◽  
Author(s):  
Thomas F. Johnson ◽  
Harry Y. C. Wong
Keyword(s):  
Plasma Protein ◽  
Plasma Proteins ◽  
Serum Proteins ◽  
College Men ◽  
Paper Electrophoresis ◽  
Acute Effects ◽  
Competitive Swimming ◽  
Protein Levels ◽  
Significant Difference ◽  
Beta Globulin

The major concern of this investigation was to observe chronic and/or acute effects of a typical intercollegiate training and competitive swimming program on plasma protein levels in young men. Plasma protein levels, followed over a period of 14 months in seven young college men, were analyzed by the technique of paper electrophoresis. No chronic effects in plasma proteins were observed; that is, no permanent change in plasma protein level was seen. Comparisons of data between competitive and noncompetitive seasons showed no significant difference. However, there were acute effects resulting from brief maximal exercise efforts. Albumin increased in response to participation in one or more competitive swimming events. A slight decrease was observed in the beta-globulin fraction. A similar albumin-beta pattern was seen in another group of varsity swimmers sprinting 220 yards. In both instances, the greatest deviation from pre-exercise levels was observed the next morning, some 11—20 hr later. Some of the mechanisms thought to act in causing a temporary alteration in plasma proteins during and after exercise are discussed. Submitted on October 12, 1960

scholarly journals THE PLACENTA AND PROTEIN METABOLISM

1955 ◽  
Vol 101 (6) ◽  
pp. 617-626 ◽  
Author(s):  
G. H. Whipple ◽  
R. B. Hill ◽  
R. Terry ◽  
F. V. Lucas ◽  
C. L. Yuile
Keyword(s):  
Amino Acids ◽  
Oral Administration ◽  
Plasma Protein ◽  
Protein Metabolism ◽  
Plasma Proteins ◽  
Maternal Plasma ◽  
Blue Dye ◽  
Placental Function ◽  
Length Of Gestation

Plasma proteins tagged in vivo by feeding D-L-lysine-ϵ-C14 to donor dogs have been administered to pregnant dogs by both oral and intravenous routes. A relatively small percentage of the C14 activity originally incorporated in these proteins is found to pass from mother to fetus after intravenous injection. The amount transferred tends to increase with the length of gestation period and total number of fetuses. Plasma protein labeled with I131 does not cross the placenta in the dog, but does in the rabbit. Evans blue dye does not cross the placenta of the dog. After oral administration of labeled plasma protein or lysine, C14 is transferred promptly and in considerable quantity to the fetus. Labeled plasma proteins disappear more rapidly from the circulation of pregnant than of normal dogs. This increased metabolic turnover occurs without excretion of any excess waste metabolites. The chorionic epithelium, gram for gram, is probably 2 to 3 times as active as the hepatic epithelium in protein metabolism. These findings indicate an important placental function related to maternal and fetal protein metabolism. While the placenta utilizes maternal plasma proteins and amino acids, in a quantitative sense the latter appear to supply the major nitrogen needs of the growing fetus.

An Evaluation of a Plasma Fractionation System

1960 ◽  
Vol 4 (01) ◽  
pp. 031-044
Author(s):  
George Y. Shinowara ◽  
E. Mary Ruth
Keyword(s):  
Biological Activity ◽  
Ionic Strength ◽  
Plasma Proteins ◽  
High Concentration ◽  
Significant Evidence ◽  
Native Proteins ◽  
Mobility Data ◽  
Fractionation Procedure ◽  
The Individual ◽  
Low Ionic Strength

SummaryFour primary fractions comprising at least 97 per cent of the plasma proteins have been critically appraised for evidence of denaturation arising from a low temperature—low ionic strength fractionation system. The results in addition to those referable to the recovery of mass and biological activity include the following: The high solubilities of these fractions at pH 7.3 and low ionic strengths; the compatibility of the electrophoretic and ultracentrifugal data of the individual fractions with those of the original plasma; and the recovery of hemoglobin, not hematin, in fraction III obtained from specimens contaminated with this pigment. However, the most significant evidence for minimum alterations of native proteins was that the S20, w and the electrophoretic mobility data on the physically recombined fractions were identical to those found on whole plasma.The fractionation procedure examined here quantitatively isolates fibrinogen, prothrombin and antithrombin in primary fractions. Results have been obtained demonstrating its significance in other biological systems. These include the following: The finding of 5 S20, w classes in the 4 primary fractions; the occurrence of more than 90 per cent of the plasma gamma globulins in fraction III; the 98 per cent pure albumin in fraction IV; and, finally, the high concentration of beta lipoproteins in fraction II.

scholarly journals Human serum protein fractionation by gel filtration

1970 ◽  
Vol 118 (5) ◽  
pp. 869-873 ◽  
Author(s):  
T. Freeman ◽  
J. Smith
Keyword(s):  
Human Serum ◽  
Serum Protein ◽  
Serum Proteins ◽  
Gel Filtration ◽  
Protein Fractionation ◽  
Logical Approach ◽  
Complex Protein ◽  
Human Serum Protein ◽  
Immunological Technique ◽  
Current Terminology

The development of a quantitative immunological technique using polyvalent antiserum permits a more logical approach to the fractionation of complex protein mixtures. In this study whole serum was separated by conventional gel filtration and the fractions obtained were analysed. This demonstrates over 60 immunologically distinct serum proteins. Because the current terminology is inadequate to describe this number of proteins, a temporary numerical nomenclature has been used.

scholarly journals BARTONELLA BODIES IN THE BLOOD OF A NON-SPLENECTOMIZED DOG

1935 ◽  
Vol 62 (3) ◽  
pp. 353-258 ◽  
Author(s):  
James B. McNaught ◽  
Francis M. Woods ◽  
Virgil Scott
Keyword(s):  
Intravenous Injection ◽  
Plasma Protein ◽  
Whole Blood ◽  
Protein Level ◽  
Plasma Proteins ◽  
Basal Diet ◽  
Blood Stream ◽  
Inhibitory Effect ◽  
The Many ◽  
Plasma Protein Level

A non-splenectomized dog, on a vitamin-adequate basal diet, in the course of a plasmapheresis experiment, developed an uncontrollable anemia associated with the presence of bodies in or on the erythrocytes, indistinguishable from the descriptions of Bartonella canis. The normal plasma protein level of 7.3 per cent was reduced to 4.1 per cent by diet and the removal of 5354 ml. of whole blood in 33 bleedings. The Bartonella infection was transferred to a splenectomized dog by an intravenous injection of whole blood. Each animal was apparently sterilized by one injection of neoarsphenamine equivalent to 15 mg. per kilo weight. It is possible that the spleen liberates some substance into the blood stream which has an inhibitory effect upon a latent Bartonella infection and that this protective substance was diminished by the many bleedings associated with the lowering of plasma proteins in the non-splenectomized dog and was lacking in the inoculated splenectomized dog.

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