Immunoassay technique for specific immunoglobulin E
OwnerMichael J. Reid, J. Michael Kwasnicki +1inventors
InventorsMichael J. Reid, J. Michael Kwasnicki, Nai-Kong V. Cheung
Life of this patent
Estimating the term…
Abstract
Using monoclonal mouse anti-human IgE antibodies, the microtiter solid phase radioimmunoassay (MSPRIA) and the enzyme-linked immunoabsorbent assay (EILSA) were modified to quantitatively measure honeybee venom (HBV) and perennial rye grass (PRG) specific immunoglobulin E (sIgE). By using a novel dilution, serial transfer and cummulative counting technique, the inhibiting and interfering effects of specific immunoglobulin G (sIgG) in the assay of (sIgE) can be reduced and the quantitative determination of (sIgE) accomplished with precision and reliability.
Claims5
A method for the quantitative determination of immunoglobulin E in a blood serum test sample which comprises the steps of:
- (A) introducing into a polyvinyl chloride test well and incubating an immunologically reactive, specific antigen for about 60 minutes at about 37.degree. C. to effect the absorption of said antigen on the wall of said test well;
- (B) aspirating said specific antigen;
- (C) introducing into said test well and incubating a globulin-free human serum albumin for about 60 minutes at about 4.degree. C.;
- (D) washing and drying said test well;
- (E) introducing into said dry test well and incubating a blood serum test sample for from about 12 to 24 hours at about 4.degree. C. to produce a blood serum supernatant fluid;
- (F) serially transferring and incubating for from about 12 to 14 hours at about 4.degree. C. said supernatant fluid to a succeeding polyvinyl chloride test well previously subjected to steps (A) to (D);
- (G) continuing to serially transfer and incubate sequentially the said supernatant fluid to a pre-determined number of succeeding test wells each of which was previously subjected to steps (A) to (D);
- (H) discarding said final supernatant fluid;
- (I) introducing into each of said predetermined number of said test wells and incubating for from about 12 to 24 hours at about 4.degree. C. a labeled, monoclonal, mouse, anti-human, immunoglobulin E antibody;
- (J) quantitatively measuring and assaying the amount of specific immunoglobulin E in each of said predetermined number of test wells to provide a cumulative total content of specific immunologlobulin E in said blood serum test sample.
Dependencies are inferred from the claim text.