Detection of hepatitis B surface antigen (or antibody to same) with labeled synthetic peptide

Claim 2

A process for the detection of antibodies to Hepatitis B in a sample which comprises: a. contacting said sample with a solid substrate coated with an unlabeled synthetic peptide comprising the following sequence of amino acids: Pro Ser Cys Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser, which peptide is recognized by and bound by the antibodies suspected of being present in said sample, incubating and washing said contacted sample, b. contacting the incubated, washed mass obtained from step a. above with a radiolabeled or enzyme labeled antibody to human or animal immunoglobulin, incubating and washing the contacted sample, and c. determining the radioactivity or enzymatic activity of the resultant mass of step b. above.

Claim 3

A process for the detection of the presence of an antibody to Hepatitis B in a sample which comprises: a. contacting said sample with a radiolabeled or enzyme labeled synthetic peptide comprising the following sequence of amino acids: Pro Ser Cys Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser, which peptide is recognized by and bound by the antibody suspected of being present in said sample, incubating and washing said contacted sample, b. adding Staphylococci bearing Protein A to said contacted sample, incubating and centrifuging the resultant mass, c. washing the resultant mass and d. determining the radioactivity or enzymatic activity of the resultant pelleted bacteria.

Claim 4

A process for the detection of the presence of an antibody to Hepatitis B in a sample which comprises: a. contacting said sample with a radiolabeled or enzyme labeled synthetic peptide comprising the following sequence of amino acids: Pro Ser Cys Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser, which peptide is recognized by and bound by the antibody suspected of being present in said sample, incubating and washing said contacted sample, b. adding Staphylococci bearing Protein A to said contacted sample, incubating and centrifuging the resultant mass, c. washing the resultant mass and d. determining the radioactivity or enzymatic activity of the resultant pelleted bacteria.

Claim 5

A process for detection of the presence of an antibody to Hepatitis B or Hepatitis B surface antigen in a sample which comprises a. contacting a first portion of a solid substrate coated with antibody with a mixture of said sample and a radiolabeled or enzyme labeled synthetic peptide having no more than 60 amino acids in the chain of the peptide, which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by the antibody in said solid substrate, incubating and washing said contacted first portion, to obtain a first resultant composition, wherein the peptide comprises the following sequence of amino acids: Pro Ser Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser; b. in a control, contacting a second portion of said solid substrate coated with said antibody with the same amount of said radiolabeled or enzyme labeled peptide as in step a. above, incubating and washing said contacted second portion, to obtain a second resultant composition, c. detecting the radioactivity or enzymatic activity of said first resultant composition from step a. above, and d. determining the radioactivity or enzymatic activity of said second resultant composition from step b. above, and e. comparing activities from steps c. and d. above, such that if the radioactivity or enzymatic activity from step c. is less than from step d., then the sample is positive for the antibody in the sample.

Claim 6

A process for detection of the presence of an antibody to Hepatitis B or Hepatitis B surface antigen in a sample which comprises a. contacting a first portion of a solid substrate coated with antibody with a mixture of said sample and a radiolabeled or enzyme labeled synthetic peptide having no more than 60 amino acids in the chain of the peptide, which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by the antibody in said solid substrate, incubating and washing said contacted first portion, to obtain a first resultant composition, wherein the peptide comprises the following sequence of amino acids: Pro Ser Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser; b. in a control, contacting a second portion of said solid substrate coated with said antibody with the same amount of said radiolabeled or enzyme labeled peptide as in step a. above, incubating and washing said contacted second portion, to obtain a second resultant composition, c. detecting the radioactivity or enzymatic activity of said first resultant composition from step a. above, and d. determining the radioactivity or enzymatic activity of said second resultant composition from step b. above, and e. comparing activities from steps c. and d. above, such that if the radioactivity or enzymatic activity from step c. is less than from step d., then the sample is positive for the antibody in the sample.

Claim 7

A process for detecting the presence of Hepatitis B surface antigen in a sample which comprises: a. contacting a first portion of a composition containing an antibody to said antigen with a mixture of said sample and a radiolabeled or enzyme labeled synthetic peptide having no more than 60 amino acids in the chain of the peptide, which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by said antibody, incubating and washing said first portion, wherein the peptide comprises the following sequence of amino acids: Pro Ser Cys Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser, b. contacting a second portion of said composition containing antibody with the same amount of said radiolabeled or enzyme labeled peptide in an antigen free control, incubating and washing said second portion, c. adding the same amount of Staphylococci bearing protein A to each of the compositions of steps a. and b. above, incubating both of said compositions, centrifuging each of said compositions and separating the supernatant liquid from the solids therein, d. determining the radioactivity or enzymatic activities of each of the resultant compositions from step c, above, and e. comparing the respective radioactivity or enzymatic activity from step d. above such that if the activity for the resultant composition containing the first portion is less than the activity for the resultant composition containing the second portion, then the sample contains antigen.

Claim 8

A process for detecting the presence of Hepatitis B surface antigen in a sample which comprises: a. contacting a first portion of a composition containing an antibody to said antigen with a mixture of said sample and a radiolabeled or enzyme labeled synthetic peptide having no more than 60 amino acids in the chain of the peptide, which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by said antibody, incubating and washing said first portion, wherein the peptide comprises the following sequence of amino acids: Pro Ser Cys Cys Cys Thr Lys Pro Thr or Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser, b. contacting a second portion of said composition containing antibody with the same amount of said radiolabeled or enzyme labeled peptide in an antigen free control, incubating and washing said second portion, c. adding the same amount of Staphylococci bearing protein A to each of the compositions of steps a. and b. above, incubating both of said compositions, centrifuging each of said compositions and separating the supernatant liquid from the solids therein, d. determining the radioactivity or enzymatic activities of each of the resultant compositions from step c, above, and e. comparing the respective radioactivity or enzymatic activity from step d. above such that if the activity for the resultant composition containing the first portion is less than the activity for the resultant composition containing the second portion, then the sample contains antigen.

Claim 9

A process for the detection of antibodies to Hepatitis B surface antigen in a sample which comprises: a. contacting said sample with a solid substrate coated with an unlabeled synthetic peptide having no more than 60 amino acids in the chain of the peptide which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by the antibody suspected of being present in the sample, incubating and washing said contacted sample, wherein the peptide comprises the following sequence of amino acids: Cys Cys Thr Lys Pro Thr Asp Gly Asn Cys Thr Cys, b. contacting the incubated, washed mass obtained from step a. above with: a radiolabeled or enzyme labeled peptide having less than 60 amino acids in the chain of the peptide, which amino acids mimic said antigenic determinant or determinants, which peptide is recognized by and bound by the antibody suspected of having been present in said sample, said peptide comprising the following sequence of amino acids: Cys Cys thr Lys Pro Thr Asp Gly Asn Cys Thr Cys, or with a radiolabeled or enzyme labeled antibody to human or animal immunoglobulin, incubating and washing the resultant mass, and c. determining the radioactivity or enzymatic activity of the resultant mass of step b. above.

Claim 10

A process for the detection of antibodies to Hepatitis B surface antigen in a sample which comprises: a. contacting said sample with a solid substrate coated with an unlabeled synthetic peptide having no more than 60 amino acids in the chain of the peptide which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by the antibody suspected of being present in the sample, incubating and washing said contacted sample, wherein the peptide comprises the following sequence of amino acids: Cys Cys Thr Lys Pro Thr Asp Gly Asn Cys Thr Cys, b. contacting the incubated, washed mass obtained from step a. above with: a radiolabeled or enzyme labeled peptide having less than 60 amino acids in the chain of the peptide, which amino acids mimic said antigenic determinant or determinants, which peptide is recognized by and bound by the antibody suspected of having been present in said sample, said peptide comprising the following sequence of amino acids: Cys Cys thr Lys Pro Thr Asp Gly Asn Cys Thr Cys, or with a radiolabeled or enzyme labeled antibody to human or animal immunoglobulin, incubating and washing the resultant mass, and c. determining the radioactivity or enzymatic activity of the resultant mass of step b. above.

Claim 11

A process for the detection of antibodies to Hepatitis B surface antigen in a sample which comprises: a. contacting said sample with a radiolabeled or enzyme labeled synthetic peptide having no more than 60 amino acids in the chain of the peptide, which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by the antibody suspected of being present in said sample, incubating and washing said contacted sample, wherein the peptide comprises the following sequence of amino acids: Cys Cys Thr Lys Pro Thr Asp Gly Asn Cys Thr Cys, b. adding Staphylococci bearing Protein A to said contacted sample, incubating and centrifuging the resultant mass, c. washing the resultant mass and d. determining the radioactivity or enzymatic activity of the resultant pelleted Staphylococci bacteria.

Claim 12

A process for the detection of antibodies to Hepatitis B surface antigen in a sample which comprises: a. contacting said sample with a radiolabeled or enzyme labeled synthetic peptide having no more than 60 amino acids in the chain of the peptide, which amino acids mimic at least one antigenic determinant of a natural antigen, which peptide is recognized by and bound by the antibody suspected of being present in said sample, incubating and washing said contacted sample, wherein the peptide comprises the following sequence of amino acids: Cys Cys Thr Lys Pro Thr Asp Gly Asn Cys Thr Cys, b. adding Staphylococci bearing Protein A to said contacted sample, incubating and centrifuging the resultant mass, c. washing the resultant mass and d. determining the radioactivity or enzymatic activity of the resultant pelleted Staphylococci bacteria.

Claim 13

A process for detecting the presence of a Hepatitis B surface antigen in a sample which comprises: a. contacting a first portion of a solid substrate coated with antibody which is an antibody to the antigen suspected in same sample with a mixture of said sample and a radiolabeled or enzyme labeled synthetic peptide selected from the group consisting of ##STR10## which peptide is recognized by and bound by the antibody in said solid substrate, incubating and washing said contacted first portion, to obtain a first resultant composition, b. in a control, contacting a second portion of said solid substrate coated with said antibody with the same amount of said radiolabeled or enzyme labeled peptide as in step a. above, incubating and washing said contacted second portion, to obtain a second resultant composition, c. detecting the radioactivity or enzymatic activity of said first resultant composition from step a. above, d. determining the radioactivity or enzymatic activity of said second resultant composition from step b. above, and e. comparing activities from steps c. and d. above, such that if the radioactivity or enzymatic activity from step c. is less than from step d., then the sample is positive for the antigen in the sample.

Claim 14

A process for detecting the presence of a Hepatitis B surface antigen in a sample which comprises: a. contacting a first portion of a solid substrate coated with antibody which is an antibody to the antigen suspected in same sample with a mixture of said sample and a radiolabeled or enzyme labeled synthetic peptide selected from the group consisting of ##STR10## which peptide is recognized by and bound by the antibody in said solid substrate, incubating and washing said contacted first portion, to obtain a first resultant composition, b. in a control, contacting a second portion of said solid substrate coated with said antibody with the same amount of said radiolabeled or enzyme labeled peptide as in step a. above, incubating and washing said contacted second portion, to obtain a second resultant composition, c. detecting the radioactivity or enzymatic activity of said first resultant composition from step a. above, d. determining the radioactivity or enzymatic activity of said second resultant composition from step b. above, and e. comparing activities from steps c. and d. above, such that if the radioactivity or enzymatic activity from step c. is less than from step d., then the sample is positive for the antigen in the sample.

Claim 15

A process for the detection of Hepatitis B surface antigen in a sample according to claim 7 wherein said synthetic peptide is enzyme labeled.