Preparation of optically clear lyophilized human serum for use in preparing control material.

1976 ◽  
Vol 22 (4) ◽  
pp. 456-460 ◽  
Author(s):  
G J Proksch ◽  
D P Bonderman
Keyword(s):  
Human Serum ◽  
Dextran Sulfate ◽  
Raw Materials ◽  
Divalent Metal ◽  
Divalent Metal Ions ◽  
Quality Control Material ◽  
Control Material ◽  
Acid Citrate Dextrose ◽  
Citrate Dextrose ◽  
Citrate Phosphate

Abstract We describe a simple, rapid process--which includes the specific precipitation of pre-beta and beta-lipoproteins with dextran sulfate and divalent metal ions--for preparing an optically clear human serum that retains its clarity upon reconstitution with water after having been frozen and lyophilized. Such serum contains the normal constituents of human serum, except for the removed lipoproteins. The process causes no apparent interference with results of analyses for 22 of the more commonly measured constituents. Fresh or aged pooled serum or blood-bank plasma containing acid-citrate-dextrose or citrate-phosphate-dextrose are equally suitable as raw materials. This stabilized serum is an excellent matrix for use in preparing standards and quality-control material for assay of components of human serum.

Assembly of particles from southern bean mosaic virus and sowbane mosaic virus components

1977 ◽  
Vol 55 (16) ◽  
pp. 2274-2277 ◽  
Author(s):  
J. H. Tremaine ◽  
W. P. Ronald
Keyword(s):  
Mosaic Virus ◽  
Dextran Sulfate ◽  
Ethylenediaminetetraacetic Acid ◽  
Sodium Dodecyl ◽  
Divalent Metal ◽  
Spherical Particles ◽  
Divalent Metal Ions ◽  
Southern Bean Mosaic Virus ◽  
Sodium Dextran Sulfate ◽  
Protein Components

Southern bean mosaic virus (SBMV) and sowbane mosaic virus (SoMV) were each dissociated into RNA and protein components in neutral pH buffers containing ethylenediaminetetraacetic acid and 1 M NaCl. The assembly of SBMV particles and of SoMV RNA in SBMV protein was accomplished by dialysis of the virus components into low molarity buffers containing divalent metal ions. Some of these particles were stable in 1% sodium dodecyl sulfate (SDS) and had sedimentation and electrophoretic properties identical with untreated virus. Spherical particles were also assembled with either SBMV or SoMV RNA in SoMV protein and with sodium dextran sulfate in either SBMV or SoMV protein, but these particles were not stable in 1% SDS.

Studies on a ph related zone phenomenon in erythrocytes

1978 ◽  
Vol 36 (2) ◽  
pp. 296-297
Author(s):  
Alan H. Timme
Keyword(s):  
Alcian Blue ◽  
Room Temperature ◽  
Unit Membrane ◽  
Paired Samples ◽  
Cacodylate Buffer ◽  
Charged Membrane ◽  
Acid Citrate Dextrose ◽  
Citrate Dextrose ◽  
Citrate Phosphate ◽  
Positively Charged

In 1967 Baker reported the occurrence of a zone of dense material adjacent to the inner leaflet of the unit membrane of erythrocytes exposed to buffers with a pH 4.5. It was postulated that this arose because the haemoglobin, positively charged at that pH, was attracted to the negatively charged membrane.In an attempt to elucidate the mechanisms underlying the formation of the zone, fresh human erythrocytes were washed for 15 minutes at room temperature in citrate phosphate buffers in which the pH ranged from 4.0 - 7.4, or in a standard preservative solution, Acid Citrate Dextrose (ACD), pH 5.0. Paired samples were subsequently washed in buffer at pH 7.4 for 10 minutes to assess the possible reversibility of any changes produced. The cells were fixed in 2.5% glutaraldehyde in 0.1 m cacodylate buffer at the same pH as the washing buffer for one hour and then processed in two ways: Some were post-fixed in OSO4, The cells were stained with 0.5 - 1% Alcian Blue 8 G S in cacodylate buffer pH 7.0 for 15 - 30 minutes.

Influence of matrix on concentrations of somatotropin measured in serum with commercial immunoradiometric assays.

1989 ◽  
Vol 35 (7) ◽  
pp. 1423-1426 ◽  
Author(s):  
R A Felder ◽  
R W Holl ◽  
P Martha ◽  
G Bauler ◽  
P Hellman ◽  
...  
Keyword(s):  
Quality Control ◽  
Human Serum ◽  
Horse Serum ◽  
Human Growth ◽  
Reference Intervals ◽  
Genetically Engineered ◽  
Quality Control Material ◽  
Serum Samples ◽  
Control Material ◽  
Human Pituitary

Abstract Using immunoradiometric assays (IRMAs) from Hybritech Inc. (H) and Nichols Institute Diagnostics (ND), we measured somatotropin (human growth hormone, hGH) in serum samples obtained every 20 min for 24 h from 10 prepubertal subjects with short stature. Results obtained with the ND reagents were 2.74 times greater than those obtained with the H reagents (P = 0.00001, r = 0.94, SEE = 3.9, n = 720). We therefore compared the IRMAs with the standard hGH RIA from the National Institutes of Health (NIH) National Hormone and Pituitary Program, using the genetically engineered hGH preparations (from Genentech Inc.) 22-kDa hGH and methionated 20-kDa hGH. We also assayed human pituitary hGH (NIH, lot no. AFP-4793B). Each hGH preparation was diluted in three diluent buffer systems: horse serum from H and from ND, and human serum. The RIA and H-IRMA gave superimposable standard curves for all hGH preparations in each diluent. The methionated 20-kDa hGH was not detected in the H-assay. Use of human serum matrix in the ND-IRMA shifted the standard curve as compared with the horse-serum matrix, giving equivalent binding at lower concentrations; i.e., serum hGH was overestimated in samples assayed against standards diluted in horse serum. Quality-control materials (Ciba-Corning) yielded disparate results in all three assays, yet human serum pools containing hGH gave similar results in the H and the NIH assays, and higher values in ND. When a human serum standard was used in the ND assay, both IRMAs gave similar results to the RIA assay for human serum samples. Reference intervals for hGH should be determined by each analytical laboratory, to prevent misdiagnosis of patients. Furthermore, quality-control material should be of human origin, because commercially supplied quality-control material does not react the same as human serum in some hGH assays.

scholarly journals Changes in intracellular Mg adenosine triphosphate and ionized Mg2+ during blood storage: detection by 31P nuclear magnetic resonance spectroscopy

Blood ◽  
1985 ◽  
Vol 65 (6) ◽  
pp. 1526-1530 ◽  
Author(s):  
JL Bock ◽  
B Wenz ◽  
RK Gupta
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Adenosine Triphosphate ◽  
Resonance Spectroscopy ◽  
Blood Storage ◽  
31P Nuclear Magnetic Resonance ◽  
Acid Citrate Dextrose ◽  
Citrate Dextrose ◽  
Citrate Phosphate ◽  
Nuclear Magnetic

Abstract 31P nuclear magnetic resonance (NMR) spectroscopy was used to measure changes in intra-erythrocyte Mg adenosine triphosphate (MgATP) and free Mg2+ during blood storage at 4 degrees C in standard citrate preservation media. The extent of Mg2+ complexation of ATP and the concentration of free Mg2+ were measured from the Mg2+-dependent chemical shift differences, at 22 degrees C, between the P beta and P alpha resonances of intracellular ATP. This difference changed from 721.0 +/- 1.4 Hz (mean +/- SE) on the day of collection to 741.0 +/- 3.4 Hz after three to seven days and 774.0 +/- 2.8 Hz after 11 to 40 days storage in either acid-citrate-dextrose (ACD) or citrate-phosphate- dextrose-adenine (CPDA-1). Changes in intracellular pH, detected from shifts in the intracellular Pi resonance, averaged 0.27 units after 11 to 40 days of storage. These data indicate a sizable decrease in the extent of Mg2+ complexation of ATP, and a decrease by a factor of 2.6 in free Mg2+, during the shelf-life of blood stored in ACD or CPDA-1.

Density Increase and Ageing of Erythrocytes in Stored Blood

1989 ◽  
Vol 17 (5) ◽  
pp. 461-466 ◽  
Author(s):  
M. Rocchigiani ◽  
M. Pescaglini ◽  
S. Sestini ◽  
V. Micheli ◽  
C. Ricci
Keyword(s):  
Dehydrogenase Activity ◽  
Whole Blood ◽  
Storage Time ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Acid Citrate Dextrose ◽  
Citrate Dextrose ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Stored Blood ◽  
Citrate Phosphate

An increase in the density of erythrocytes was observed after storage of whole blood for 30 days at 4°C in either acid citrate – dextrose or citrate – phosphate – dextrose – adrenaline. Glucose- 6-phosphate dehydrogenase activity in unfractionated red blood cell lysates did not vary with the storage time. Enzyme activity in the lighter fraction separated by density gradient centrifugation was higher than that in heavier fractions. The decline in glucose-6-phosphate dehydrogenase activity with density was less marked after storage of whole blood for 30 days. It is suggested that density modifications are not related to the ageing of erythrocytes and additional mechanisms may be involved.

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