Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals
Patent
1990-11-15
1992-05-19
Kepplinger, Esther L.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
435 792, 435 7021, 43524027, 435970, 435 74, 436528, 436548, 436805, 436810, 53038825, 5303882, 5303893, 5303891, G01N 33543
Patent
active
051148633
ABSTRACT:
A double antibody sandwich immunosorbent assay for detecting alpha-1-antitrypsin (.alpha..sub.1 AT) comprising the steps of: (1) reacting (a) a first-species-of-animal antibody to .alpha..sub.1 AT with (b) a solid substrate; (2) reacting (a) .alpha..sub.1 AT in a sample suspected of containing .alpha..sub.1 AT with (b) the first-species-of animal polyclonal antibody to .alpha..sub.1 AT; (3) reacting (a) a second-species-of-animal antibody to .alpha..sub.1 AT with (b) the .alpha..sub.1 AT in a sample suspected of containing .alpha..sub.1 AT; (4) reacting (a) a labelled antibody comprising a first-species-of-animal antibody to an antibody from a second species of animal with (b) the second-species-of-animal antibody to .alpha..sub.1 AT; and (5) detecting the amount of labelled first-species-of-animal antibody to antibody from the second species of animal that reacted with the second-species-of-animal polyclonal antibody to .alpha..sub.1 AT; provided that the first species of animal and the second species of animal are not the same in a given assay; and provided that steps (1) to (4) can be conducted in any order so as to result in a complex comprising: (i) the substrate complexed to (ii) the first-species-of-animal antibody to .alpha..sub.1 AT complexed to (iii) the .alpha..sub.1 AT complexed to (iv) the second-species-of-animal antibody to .alpha..sub.1 AT complexed to (v) the labelled antibody. The antibodies used in the assay can be polyclonal antibodies or monoclonal antibodies. A kit for detecting .alpha..sub.1 AT comprising reagents for conducting the above described assays. A monoclonal antibody having specific reactivity to .alpha..sub.1 AT, the antibody being further characterized in that it exhibits substantially no cross reactivity with albumin. A hybridoma that produces the above described monoclonal antibody.
REFERENCES:
patent: 4590157 (1986-05-01), Chandler et al.
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Michalski et al., "A Modified Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay for Measurement of .alpha.-1-Antitrypsin in Biologic Fluids", J. Immun. Methods, 83 (1985) 101-102.
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Gallaher William R.
McCombs Candace C.
Michalski Joseph P.
Thompson James J.
Board of Supervisors of Louisiana State University & Agricultura
Kepplinger Esther L.
Spiegel Carol A.
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