Pretreatment method for carcinoembryonic antigen assay

Claim 2

In a method of measuring the level of carcinoembryonic antigen (CEA) in a specimen by pretreating with perchloric acid to dissociate the CEA material from binding proteins by precipitating said binding proteins, separating said precipitate from the CEA-containing supernatant, and measuring the level of CEA wherein the improvement comprises eliminating the necessity of dialysis to purify the perchloric acid treated specimen by: (a) adding to said CEA-containing supernatant a buffered source of potassium ions to precipitate potassium perchlorate; and (b) separating said potassium perchlorate precipitate to yield a CEA-containing supernatant ready for assay.

Claim 3

A method as described in claim 1 wherein the specimen is human plasma or serum.

Claim 4

A method as described in claim 1 wherein the buffered source of potassium ions yields a pH of the treated sample in the range of about 4.5 to 8.0.

Claim 5

A method as described in claim 1 wherein the buffered source of potassium ions is a mixture of KOH and K 2 HPO 4 .

Claim 6

A method as described in claim 1 wherein said CEA-containing supernatant is assayed using a solid phase radioimmunoassay.

Claim 7

A method as described in claim 1 wherein said neutralized CEA-containing supernatant is assayed using an immunoassay.

Claim 8

A relatively rapid, sensitive assay for measuring the CEA level in a specimen, said assay characterized by tolerance toward changes in system pH and ionic strength and comprising the steps of: (a) pretreating a specimen of blood with a perchloric acid; (b) centrifuging the mixture from step (a) to obtain a CEA-containing supernatant. (c) adding to said supernatant a buffered source of potassium ions to precipitate potassium perchlorate; (d) separating without dialysis, said salt to yield a CEA-containing supernatant; (e) incubating a support having a coating of CEA antibody thereon with a known amount of said supernatant from step (d), to thereby bind any CEA present in said supernatant to said coating of antibody; (f) washing said thus coated support; (g) incubating said washed support with a known amount of labeled CEA antibody to produce a layer of labeled antibody bound to the CEA on the antibody coated support; (h) washing the labeled, coated support of step (g); and (i) detecting quantitatively the amount of labeled antibody on the washed, labeled and coated support to thereby obtain information directly related to the level of CEA present in the specimen.

Claim 9

A relatively rapid, sensitive assay for measuring the CEA level in a specimen, said assay characterized by tolerance toward changes in system pH and ionic strength and comprising the steps of: (a) pretreating a specimen of blood with a perchloric acid; (b) centrifuging the mixture from step (a) to obtain a CEA-containing supernatant. (c) adding to said supernatant a buffered source of potassium ions to precipitate potassium perchlorate; (d) separating without dialysis, said salt to yield a CEA-containing supernatant; (e) incubating a support having a coating of CEA antibody thereon with a known amount of said supernatant from step (d), to thereby bind any CEA present in said supernatant to said coating of antibody; (f) washing said thus coated support; (g) incubating said washed support with a known amount of labeled CEA antibody to produce a layer of labeled antibody bound to the CEA on the antibody coated support; (h) washing the labeled, coated support of step (g); and (i) detecting quantitatively the amount of labeled antibody on the washed, labeled and coated support to thereby obtain information directly related to the level of CEA present in the specimen.

Claim 10

A relatively rapid, sensitive assay for measuring the CEA level in a specimen, said assay characterized by tolerance toward changes in system pH and ionic strength and comprising the steps of: (a) pretreating a specimen of blood with a perchloric acid; (b) centrifuging the mixture from step (a) to obtain a CEA-containing supernatant; (c) adding to said supernatant a buffered source of potassium ions to precipitate potassium perchlorate; (d) separating without dialysis, said salt to yield a CEA-containing supernatant; (e) incubating a support having a coating of CEA antibody thereon with a known amount of said supernatant from step (d) and with a known amount of labeled CEA to thereby permit competition between the labeled CEA and any CEA present in said supernatant for antibody on the support; (f) washing the labeled, coated support of step (e); and (g) detecting quantitatively the amount of labeled CEA on the washed, labeled and coated support to thereby obtain information directly related to the level of CEA present in the specimen.

Claim 11

A relatively rapid, sensitive assay for measuring the CEA level in a specimen, said assay characterized by tolerance toward changes in system pH and ionic strength and comprising the steps of: (a) pretreating a specimen of blood with a perchloric acid; (b) centrifuging the mixture from step (a) to obtain a CEA-containing supernatant; (c) adding to said supernatant a buffered source of potassium ions to precipitate potassium perchlorate; (d) separating without dialysis, said salt to yield a CEA-containing supernatant; (e) incubating a support having a coating of CEA antibody thereon with a known amount of said supernatant from step (d) and with a known amount of labeled CEA to thereby permit competition between the labeled CEA and any CEA present in said supernatant for antibody on the support; (f) washing the labeled, coated support of step (e); and (g) detecting quantitatively the amount of labeled CEA on the washed, labeled and coated support to thereby obtain information directly related to the level of CEA present in the specimen.