scholarly journals Separation of antigens by immunological specificity. Studies on the desorption of homologous antigen from disulphide-linked antibody immunosorbents

1970 ◽  
Vol 117 (1) ◽  
pp. 49-55 ◽  
Author(s):  
J. W. Chidlow ◽  
J. Stephen ◽  
H. Smith
Keyword(s):  
Hydrochloric Acid ◽  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Cellulose Acetate ◽  
Cellulose Acetate Electrophoresis ◽  
Acid Buffer ◽  
Homologous Antigen ◽  
Immunological Specificity

1. Glycine–hydrochloric acid buffer, pH2.2, desorbed 131I-labelled human serum albumin (100%), lysozyme (100%), ovalbumin (90%), fluorescent ovalbumin (50–60%) and fluorescent human γ-globulin (20%) from their respective homologous disulphide-linked antibody immunosorbents; reasons are suggested for the low recoveries of the fluorescently labelled proteins. 2. Approx. 40% of the recovered 131I-labelled human serum albumin and fluorescent ovalbumin was desorbed above pH6.0, but lysozyme was not eluted until the pH was 3.0 or below. 3. In all cases where high recoveries of antigen were obtained, the immunosorbents could be regenerated and recycled at least four times with full retention of specificity and minimal diminution of capacity. 4. The desorbed antigens were unchanged when compared with the original antigens by quantitative precipitin, specificradioactivity, fluorescent and enzymic analyses and by cellulose acetate electrophoresis. 5. Desorption of antigen with a variety of reagents was investigated. These reagents were less satisfactory than glycine–hydrochloric acid.

Isolation of Albumin from Human Serum by Means of Trichloroacetic Acid and Ethanol

1968 ◽  
Vol 14 (1) ◽  
pp. 22-30 ◽  
Author(s):  
Takuzo Iwata ◽  
Hiromi Iwata ◽  
James F Holland
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Cellulose Acetate ◽  
Present Method ◽  
Trichloroacetic Acid ◽  
Aqueous Suspension ◽  
Final Concentration ◽  
Cellulose Acetate Electrophoresis ◽  
Excellent Reproducibility

Abstract A rapid method for nearly quantitative isolation of human serum albumin is described. The present technic (1) is a modification of the Schwert method (2). In the present method 4 ml. of serum is precipitated with trichloroacetic acid in a final concentration of 5% (w/v). The precipitate is washed with TCA, and after aqueous suspension, ethanol is added to a final concentration of 80%. The undissolved globulin is washed with ethanol and the solubilized albumin used for quantification, characterization, and isotope counting. Recovery of added human serum albumin was approximately 98%; recovery of added 131I albumin was 90%. Cellulose acetate electrophoresis showed a single band, and agar diffusion produced one precipitate line. Immunoelectrophoresis with anti-human serum revealed one or two very faint arcs in the α-, or α- and β-globulin regions, in addition to the major arc of albumin. The method has shown excellent reproducibility in replicate analyses of normal and pathologic serums.

scholarly journals Separation of antigens by immunological specificity. Use of disulphide-linked antibodies as immunosorbents

1966 ◽  
Vol 101 (3) ◽  
pp. 717-720 ◽  
Author(s):  
J Stephen ◽  
RGC Gallop ◽  
H Smith
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Gamma Globulin ◽  
High Capacity ◽  
High Specificity ◽  
Homologous Antigen ◽  
Immunological Specificity ◽  
Insoluble Polymers ◽  
Low Solubility

1. gamma-Globulin concentrates of antisera prepared against ovalbumin and human serum albumin were thiolated and cross-linked to form insoluble polymers. 2. These immunosorbents were of low solubility and of high capacity for homologous antigen. 3. The high specificity of these immunosorbents was demonstrated by fractionation of various binary mixtures of fluorescent ovalbumin, (131)I-labelled human serum albumin, lysozyme and ribonuclease.

scholarly journals Separation of antigens by immunological specificity. Use of cellulose-linked antibodies as immunosorbents

1966 ◽  
Vol 101 (3) ◽  
pp. 711-716 ◽  
Author(s):  
RGC Gallop ◽  
BT Tozer ◽  
J Stephen ◽  
H Smith
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Gamma Globulin ◽  
High Specificity ◽  
Immunological Specificity

1. Immunosorbents were prepared by coupling activated aminocellulose with the gamma-globulin concentrates of antisera prepared against ovalbumin and human serum albumin. 2. The immunosorbents were low in solubility, but high in capacity for homologous antigens. 3. The high specificity of these immunosorbents was demonstrated by their use in fractionating various mixtures of fluorescent ovalbumin, (131)I-labelled human serum albumin, lysozyme and ribonuclease.

scholarly journals The specificity of antisera against crystalline serum albumin

1939 ◽  
Vol 39 (2) ◽  
pp. 170-180 ◽  
Author(s):  
Muriel E. Adair ◽  
John Hamilton
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Horse Serum ◽  
Optimal Point ◽  
Homologous Antigen ◽  
Antigen Antibody

Antisera prepared in the rabbit by injection of crystalline horse-serum albumin or crystalline human-serum albumin show a considerable diminution in specificity after more than one course of six to eight injections has been given.The ratio of precipitate nitrogen to antigen nitrogen at the optimal point is smaller in antisera obtained after one course of injections than in antisera derived from subsequent bleedings.In the case of antisera obtained from first bleedings, the ratio of precipitate nitrogen to antigen nitrogen determined at the optimal point with homologous antigen is of approximately the same value in both the antigen-antibody systems studied. The increase in the values of the ratios for antisera from subsequent bleedings is also the same for both systems.

II. Lokalisation der Placenta mit 113Inm- Human-Serum-Albumin

1969 ◽  
Vol 08 (01) ◽  
pp. 15-21 ◽  
Author(s):  
K. E. Scheer ◽  
J. Heep ◽  
W. Maier-Borst ◽  
W. J. Lorenz ◽  
H. Sinn ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Vena Cava ◽  
Placenta Praevia ◽  
Wird Eine

ZusammenfassungNach tierexperimentellen Voruntersuchungen wurde die Placentographie mit trägerfreiem 113Inm -HSA als klinische Methode eingeführt. Vor Amniocentesen und bei Verdacht auf Placenta praevia werden Placentographien geschrieben. Den Schwangeren wird eine Aktivität von 500 μCi in die Cubitalvene injiziert. Die der Aktivität entsprechende Indiummenge ist kleiner als 0,1 ng. Die fetale Strahlenbelastung liegt unter lOmrad. Bei Anwendung von 113Inm-HSA entfällt eine Blockade der mütterlichen und fetalen Schilddrüsen. Die genaue Abgrenzung einer Placenta praevia wird nicht durch eine Blasenaktivität beeinträchtigt.Es wurden bisher 19 Placentalokalisationen durchgeführt. In allen Fällen konnte der Placentasitz eindeutig festgestellt werden. Bedingt durch die lange Liegezeit beim Aufnehmen eines Szintigramms kam es in zwei Fällen zu einem Vena-Cava-Kompressions-Syndrom. Zur Verhinderung dieser klinischen Zwischenfälle werden inzwischen Placentographien mit der Anger-Kamera aufgenommen. Mit Hilfe des divergierenden Kollimators konnte der gesamte Abdominalbereich erfaßt werden. Die Aufnahmezeit konnte auf 7 — 10 Minuten verkürzt werden. Die intravenöse injizierte Aktivität betrug bei dieser Methode ebenfalls 500 μCi. Der diagnostische Aussagewert der Kamerabilder ist szintigraphischen Aufnahmen gleichwertig.

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