A New Technique in Quantitative Immunohematology: Solid-Phase Kinetic Enzyme-Linked Immunosorbent Assay

Vox Sanguinis ◽  
1983 ◽  
Vol 45 (6) ◽  
pp. 440-448 ◽  
Author(s):  
S. Spitalnik ◽  
J. Cowles ◽  
M.T. Cox ◽  
D. Baker ◽  
J. Holt ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
New Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
A New Technique

A New Technique in Quantitative Immunohematology: Solid-Phase Kinetic Enzyme-Linked Immunosorbent Assay

Vox Sanguinis ◽  
1983 ◽  
Vol 45 (6) ◽  
pp. 440-448 ◽  
Author(s):  
S. Spitalnik ◽  
J. Cowles ◽  
M. T. Cox ◽  
D. Baker ◽  
J. Holt ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
New Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
A New Technique

Enzyme-linked immunosorbent assay for detection of antibodies to extractable nuclear antigens in systemic lupus erythematosus, with nylon as solid phase.

1983 ◽  
Vol 29 (5) ◽  
pp. 823-827 ◽  
Author(s):  
M Ishaq ◽  
R Ali
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Immunosorbent Assay ◽  
Lupus Erythematosus ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Prolonged Storage ◽  
Antibody Activity ◽  
Systemic Lupus ◽  
Nuclear Antigens ◽  
As Solid Phase

Abstract In this enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against extractable nuclear antigens (ENA) in sera of patients with systemic lupus erythematosus (SLE), nylon is used as solid phase for antigen binding instead of the commonly used polystyrene surface. Optimal conditions for activation of the nylon beads, antigen coating, and other relevant factors have been investigated. We compared the incidence of anti-ENA antibodies in SLE, using chromogenic and fluorogenic enzyme substrates. Of SLE patients, 54% were positive for anti-ENA antibodies when chromogenic substrate was used as compared with 68% for fluorogenic substrate. Antibody activity against Sm and RNP antigens was distinguished on the basis of ribonuclease sensitivity of the RNP antigen. The method described offers advantages such as decreased background activity, increased surface area, facility for prolonged storage of antigen-coated solid phase, and miniaturization of the assay.

A solid-phase enzyme-linked immunosorbent assay for the quantitation of human plasma alpha 1-acid glycoprotein.

1979 ◽  
Vol 25 (4) ◽  
pp. 546-549 ◽  
Author(s):  
H P Wang ◽  
C Y Chu
Keyword(s):  
Alkaline Phosphatase ◽  
Human Plasma ◽  
Immunosorbent Assay ◽  
Specific Antibody ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Normal Range ◽  
Oral Contraceptive Pills ◽  
Acid Glycoprotein ◽  
Contraceptive Pills

Abstract We describe a solid-phase enzyme-linked immunosorbent assay for alpha 1-acid glycoprotein in human plasma. Plasma samples are incubated with alkaline phosphatase-linked, purified alpha 1-acid glycoprotein in alpha 1-acid glycoprotein-specific antibody-coated polystyrene tubes. The alkaline phosphatase that becomes attached to the tube via an immunological reaction between the alpha 1-acid glycoprotein and the specific antibody is measured spectrophotometrically. This assay is accurate reproducible, simple, and economical. As little as 4 microgram of alpha 1-acid glycoprotein per liter can be detected. The normal range for alpha 1-acid glycoprotein in the plasma of healthy adults, as measured by this method, is 0.48-1.27 g/L; the range is significantly different, 0.29-0.73 g/L, for women who are taking oral contraceptive pills.

Enzyme-Linked Immunosorbent Assay For The Detection Of Alloantibodies To Factor IX In Hemophilia B

1981 ◽  
Author(s):  
K H Örstavik ◽  
I Örstavik
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
Factor Ix ◽  
Hemophilia B ◽  
Negative Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nitrophenyl Phosphate ◽  
Human Factor Ix ◽  
Coagulation Assay

A solid phase enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantitative determination of acquired inhibitors to factor IX. Wells of polystyren Micro-ELISA plates were coated with the IgG fraction of a sheep antiserum to human factor IX. After incubation with pooled normal plasma as a factor IX source, the wells were incubated with test plasma. The binding of alloantibodies to the factor IX-sheep-anti-factor IX complexes was then detected by incubation with alkaline phophatase conjugated antiserum to human IgG. As substrate was used p-nitrophenyl phosphate.Plasma samples from five patients with severe hemophilia B and acquired inhibitors to factor IX were examined. All samples gave a positive reaction in the ELISA. The titers as determined in the ELISA were in good agreement with the titers as determined in a coagulation assay (0.1-800 U/ml). Plasma from 13 patients with hemophilia B and no detectable inhibitor in a coagulation assay all gave a negative reaction in the ELISA. A negative reaction was also found in plasma from four patients with hemophilia A and acquired inhibitors to factor VIII, and in plasma from 15 healthy persons.It is concluded that the ELISA is a simple and sensitive technique for the determination of acquired inhibitors to factor IX in hemophilia B.

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