An "antibody electrode," preliminary report on a new approach in enzyme immunoassay.

1979 ◽  
Vol 25 (2) ◽  
pp. 318-321 ◽  
Author(s):  
J L Boitieux ◽  
G Desmet ◽  
D Thomas
Keyword(s):  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Solid Phase ◽  
Biological Fluids ◽  
Hepatitis B Surface Antigen ◽  
Antigen Concentration ◽  
New Approach ◽  
Solid Phase Radioimmunoassay ◽  
Reliable Procedure ◽  
Sensitive Electrode

Abstract We report preliminary experiments on a new, sensitive, and reliable procedure for enzyme immunoassay of various antigens in biological fluids. The method, developed from the biological model "hepatitis B surface-antigen/antibodies," is less time consuming than most immunochemical techniques and eliminates many inconveniences arising from use of isotopes. We use a solid-phase "sandwich" procedure, the antibodies being immobilized on gelatin membranes, and determine antigen concentration with the help of an iodide-sensitive electrode modified by fixing the active membrane onto the crystal sensor. We have established the analytical criteria of the method and compared it with the solid-phase radioimmunoassay for the surface antigen in dilution series. One-tenth microgram of antigen per liter can be reproducibly detected with our method. Use of antibody electrode should easily be extended to assay of other antigens and haptens that usually are determined by radioimmunoassay.

scholarly journals Effect of solid phase radioimmunoassay-bead method to detect hepatitis B surface antigen.

RADIOISOTOPES ◽  
1984 ◽  
Vol 33 (12) ◽  
pp. 887-890
Author(s):  
Akio YOSHIMOTO ◽  
Yoshikatsu SAKAGISHI ◽  
Jean Dean LIU ◽  
Seiei TANAKA
Keyword(s):  
Hepatitis B ◽  
Surface Antigen ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen ◽  
Solid Phase Radioimmunoassay ◽  
Bead Method

scholarly journals Solid-phase enzyme-immunoassay for detection of hepatitis B surface antigen.

1976 ◽  
Vol 29 (10) ◽  
pp. 873-879 ◽  
Author(s):  
G Wolters ◽  
L Kuijpers ◽  
J Kacaki ◽  
A Schuurs
Keyword(s):  
Hepatitis B ◽  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen

Solid-phase enzyme immunoassay for hepatitis B surface antigen.

1977 ◽  
Vol 23 (5) ◽  
pp. 813-815 ◽  
Author(s):  
R Wei ◽  
G J Knight ◽  
D H Zimmerman ◽  
H E Bond
Keyword(s):  
Hepatitis B ◽  
Horseradish Peroxidase ◽  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Room Temperature ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen ◽  
Solid Support ◽  
Controlled Pore Glass ◽  
Pore Glass

Abstract A solid-phase enzyme immunoassay is described for measuring hepatitis B surface antigen in human serum or plasma. Immunologically purified antibody labeled with horseradish peroxidase was used as the indicator. In the assay system, antibody-coated controlled-pore glass is used as a solid support and there are three sequential incubations, totaling 2 h, at room temperature. Results for serially diluted positive and reference sera compare favorably to radioimmunoassay in sensitivity and specificity.

A solid-phase enzyme immunoassay for the common and subtypic determinants of hepatitis B surface antigen with monoclonal antibodies

1986 ◽  
Vol 87 (2) ◽  
pp. 203-210 ◽  
Author(s):  
Sadakazu Usuda ◽  
Fumio Tsuda ◽  
Tohru Gotanda ◽  
Katsumi Tachibana ◽  
Motozumi Nomura ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Hepatitis B ◽  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen ◽  
The Common

A monoclonal-antibody enzyme immunoassay for detection of hepatitis B surface antigen with use of a biotin-avidin system.

1985 ◽  
Vol 31 (2) ◽  
pp. 202-205 ◽  
Author(s):  
Y S Liu ◽  
A Green
Keyword(s):  
Hepatitis B ◽  
Enzyme Immunoassay ◽  
Surface Antigen ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen ◽  
Standard Procedure ◽  
Complex Form ◽  
Antibody Complex ◽  
Covalently Linked ◽  
Viral Determinants

Abstract In this solid-phase two-site enzyme immunoassay for hepatitis B surface antigen (HBsAg), three monoclonal anti-HBs are used: 5D3 (IgM) is immobilized on plastic beads; 5C3 (IgG2a) and 5C 11 (IgG1), labeled with biotin, are used as the first conjugate. Horseradish peroxidase covalently linked to avidin is the second conjugate. First, serum or plasma is incubated with the antibody-coated bead and biotin-labeled antibodies, simultaneously, at 45 degrees C for 1 h ("stat" procedure), 3 h ("standard" procedure), or 18 h ("overnight" procedure), during which HBsAg forms a complex with the solid-phase antibody and the biotinylated antibodies. The enzyme-conjugated avidin is then bound to the biotin on the antigen-antibody complex at 45 degrees C for 15 min ("stat") or 30 min (standard and overnight procedures). The beads are incubated with enzyme-substrate solution (H2O2 and o-phenylenediamine). Color developed is measured at 492 nm. All procedures satisfied third-generation HBsAg tests required by the FDA Office of Biologics, being sensitive both to ad and ay subtypes in subnanogram amounts. The assay is reactive with adw2, adw4, adr, ayw2, ayw3, and ayr subtypes and can detect viral determinants in HBsAg-anti-HBs immune complex form. Thus it provides a sensitive, simple, and reproducible alternative to radioimmunoassay.

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