Double capture assay method employing a capillary flow device

Claim 2

A double capture immunoassay for an antigen of interest in a sample, comprising the steps of: a. combining: 1) the sample; 2) magnetic particles having affixed thereto antigen of interest; 3) nonmagnetic detectable particles having affixed thereto the antigen of interest; and 4) unbound antibody specific for the antigen of interest, to produce a reaction mixture; b. incubating the reaction mixture under conditions and for a period of time sufficient for antigen of interest to bind to antibody specific for the antigen; c. separating the magnetic particles from the nonmagnetic particles; d. capturing the nonmagnetic particles on a solid material; and e. determining the presence or absence of captured nonmagnetic particles on the solid material. f. determining the presence or absence of the antigen of interest in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the antigen of interest and the antigen of interest.

Claim 3

A double capture immunoassay of claim 1 wherein the antigen of interest is quantitated by determining the number of captured nonmagnetic particles on the solid material and relating the number of captured nonmagnetic particles present on the solid material to a predetermined quantitative relationship between the number of captured nonmagnetic particles and the quantity of antigen of interest.

Claim 4

A double capture immunoassay for an antibody of interest in a sample, comprising the steps of: a. combining: 1) the sample; 2) magnetic particles having affixed thereto antigen for which the antibody of interest is specific; and 3) nonmagnetic detectable particles having affixed thereto antigen for which the antibody of interest is specific, to produce a reaction mixture; b. incubating the reaction mixture under conditions and for a period of time sufficient for antibody of interest to bind to antigen for which it is specific; c. separating the magnetic particles from the nonmagnetic particles; d. capturing the nonmagnetic particles on a solid material; e. determining the presence or absence of captured nonmagnetic particles on the solid material. f. determining the presence or absence of the antibody of interest in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the antigen for which the antibody of interest is specific and the antibody of interest.

Claim 5

A double capture immunoassay for an antibody of interest in a sample, comprising the steps of: a. combining: 1) the sample; 2) magnetic particles having affixed thereto antigen for which the antibody of interest is specific; and 3) nonmagnetic detectable particles having affixed thereto antigen for which the antibody of interest is specific, to produce a reaction mixture; b. incubating the reaction mixture under conditions and for a period of time sufficient for antibody of interest to bind to antigen for which it is specific; c. separating the magnetic particles from the nonmagnetic particles; d. capturing the nonmagnetic particles on a solid material; e. determining the presence or absence of captured nonmagnetic particles on the solid material. f. determining the presence or absence of the antibody of interest in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the antigen for which the antibody of interest is specific and the antibody of interest.

Claim 6

A double capture immunoassay of claim 3 wherein the antibody of interest is quantitated by determining the number of captured nonmagnetic particles on the solid material and relating the number of captured nonmagnetic particles present on the solid material to a predetermined quantitative relationship between the number of captured nonmagnetic particles and the quantity of antibody of interest.

Claim 7

A double capture assay for a drug of abuse in a sample, comprising the steps of: a. combining: 1) the sample; 2) magnetic particles having affixed thereto the drug of abuse or a metabolite thereof; 3) nonmagnetic detectable particles having affixed thereto the drug of abuse or a metabolite thereof; and 4) specific binding partner for the drug of abuse or a metabolite thereof, to produce a reaction mixture; b. incubating the reaction mixture under conditions and for a period of time sufficient for the drug of abuse and the specific binding partner to bind; c. separating the magnetic particles from the nonmagnetic particles; d. capturing the nonmagnetic particles; and e. determining the presence or absence of captured particles. f. determining the presence or absence of the drug of abuse in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the drug of abuse or a metabolite thereof and the drug of abuse.

Claim 8

A double capture assay for a drug of abuse in a sample, comprising the steps of: a. combining: 1) the sample; 2) magnetic particles having affixed thereto the drug of abuse or a metabolite thereof; 3) nonmagnetic detectable particles having affixed thereto the drug of abuse or a metabolite thereof; and 4) specific binding partner for the drug of abuse or a metabolite thereof, to produce a reaction mixture; b. incubating the reaction mixture under conditions and for a period of time sufficient for the drug of abuse and the specific binding partner to bind; c. separating the magnetic particles from the nonmagnetic particles; d. capturing the nonmagnetic particles; and e. determining the presence or absence of captured particles. f. determining the presence or absence of the drug of abuse in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the drug of abuse or a metabolite thereof and the drug of abuse.

Claim 9

A double capture assay of claim 5 wherein the drug of abuse is cocaine, the metabolite thereof is benzoylecgonine, and the specific binding partner is monoclonal anti-benzoylecgonine.

Claim 10

An assay for detecting an analyte of interest which is a member of a specific binding pair in a sample, comprising the steps of: a) combining the following components: i) the sample; ii) magnetic particles having affixed thereto a first member of a specific binding pair; iii) non-magnetic detectable particles having affixed thereto a second member of a specific binding pair; and b) incubating the mixture from step a) under conditions appropriate for the binding of complementary members of a specific binding pairs; c) separating the magnetic particles, and non-magnetic particles bound thereto, from unbound non-magnetic particles; d) capturing the unbound non-magnetic particles; and e) determining the presence of captured unbound non-magnetic particles. f. determining the presence or absence of the analyte of interest in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the second member of the specific binding pair and the analyte of interest.

Claim 11

An assay for detecting an analyte of interest which is a member of a specific binding pair in a sample, comprising the steps of: a) combining the following components: i) the sample; ii) magnetic particles having affixed thereto a first member of a specific binding pair; iii) non-magnetic detectable particles having affixed thereto a second member of a specific binding pair; and b) incubating the mixture from step a) under conditions appropriate for the binding of complementary members of a specific binding pairs; c) separating the magnetic particles, and non-magnetic particles bound thereto, from unbound non-magnetic particles; d) capturing the unbound non-magnetic particles; and e) determining the presence of captured unbound non-magnetic particles. f. determining the presence or absence of the analyte of interest in the sample by relating the presence or absence of captured nonmagnetic particles to a relationship between captured nonmagnetic detectable particles having affixed thereto the second member of the specific binding pair and the analyte of interest.

Claim 12

A method of claim 7 wherein steps b), c) and d) are carried out in a capillary flow device comprising: a) a capillary track having a first and a second distal end; b) a sample receptacle in communication with the first distal end of the capillary track; and c) a liquid reservoir at the second distal end of the capillary track.

Claim 13

A method of claim 8 wherein the capillary flow device further comprises: d) a particle concentrator in communication with the second distal end of the capillary track.

Claim 14

An assay of claim 7 wherein the amount of the analyte of interest present in the sample is quantitated by determining the number of captured unbound non-magnetic particles and relating the number of captured unbound non-magnetic particles to a predetermined quantitative relationship between the number of captured non-magnetic particles and the quantity of the analyte of interest.

Claim 15

An assay of claim 7 wherein the analyte of interest is an antigen specifically reactive with an antibody, the first member of a specific binding pair is an antigen specifically reactive with the antibody and the second member of a specific binding pair is the antibody.

Claim 16

An assay of claim 7 wherein the analyte of interest is an antigen specifically reactive with an antibody, the first member of a specific binding pair is the antibody and the second member of a specific binding pair is an antigen specifically reactive with the antibody.

Claim 17

An assay of claim 7 wherein the analyte of interest is an antigen, the first member of a specific binding pair is an antibody specifically reactive with the antigen and the second member of a specific binding pair is an antibody specifically reactive with the antigen.

Claim 18

An assay of claim 7 wherein the analyte of interest is an antibody and the first member of a specific binding pair is an antigen specifically reactive with the antibody and the second member of a specific binding pair is an antigen specifically reactive with the antibody.

Claim 19

An assay of claim 7 wherein the analyte of interest is an antibody and the first member of a specific binding pair is an antigen specifically reactive with the antibody and the second member of a specific binding pair is an antibody specifically reactive with the antigen.

Claim 20

An assay of claim 7 wherein the analyte of interest is an antibody specifically reactive with an antigen and the first member of a specific binding pair is an antibody specifically reactive with the antigen and the second member of a specific binding pair is the antigen.

Claim 21

An assay for detecting an analyte of interest which is a member of a specific binding pair in a sample, comprising the steps of: a) combining the following components: i) the sample; ii) magnetic particles having affixed thereto a first member of a specific binding pair; iii) non-magnetic detectable particles having affixed thereto a second member of a specific binding pair; and iv) a third member of a specific binding pair specifically reactive with the analyte of interest; and b) incubating the mixture from step a) under conditions appropriate for binding of complementary members of specific binding pairs; c) separating the magnetic particles, and non-magnetic particles bound thereto, from unbound non-magnetic particles; d) capturing the unbound non-magnetic particles; and e) determining the presence of captured unbound non-magnetic particles. f. determining the presence or absence of the analyte of interest in the sample by relating the presence or absence of captured non-magnetic particles to a relationship between captured non-magnetic detectable particles having affixed thereto the second member of the specific binding pair and the analyte of interest.

Claim 22

An assay for detecting an analyte of interest which is a member of a specific binding pair in a sample, comprising the steps of: a) combining the following components: i) the sample; ii) magnetic particles having affixed thereto a first member of a specific binding pair; iii) non-magnetic detectable particles having affixed thereto a second member of a specific binding pair; and iv) a third member of a specific binding pair specifically reactive with the analyte of interest; and b) incubating the mixture from step a) under conditions appropriate for binding of complementary members of specific binding pairs; c) separating the magnetic particles, and non-magnetic particles bound thereto, from unbound non-magnetic particles; d) capturing the unbound non-magnetic particles; and e) determining the presence of captured unbound non-magnetic particles. f. determining the presence or absence of the analyte of interest in the sample by relating the presence or absence of captured non-magnetic particles to a relationship between captured non-magnetic detectable particles having affixed thereto the second member of the specific binding pair and the analyte of interest.

Claim 23

A method of claim 17 wherein steps b), c) and d) are carried out in a capillary flow device comprising: a) a capillary track having a first and a second distal end; b) a sample receptacle in communication with the first distal end of the capillary track; and c) a liquid reservoir at the second distal end of the capillary track.

Claim 24

A method of claim 18 wherein the capillary flow device further comprises: d) a particle concentrator in communication with the second distal end of the capillary track.

Claim 25

An assay of claim 17 wherein the amount of the analyte of interest present in the sample is quantitated by determining the number of captured unbound non-magnetic particles and relating the number of captured unbound non-magnetic particles to a predetermined quantitative relationship between the number of captured non-magnetic particles and the quantity of the analyte of interest.

Claim 26

An assay of claim 17 wherein the analyte of interest is an antigen reactive with an antibody, the first member of a specific binding pair is an antigen specifically reactive with the antibody, the second member of a specific binding pair is antigen specifically reactive with the antibody and the third member of a specific binding pair is the antibody.

Claim 27

An assay of claim 17 wherein the analyte of interest is an antibody specifically reactive with an antigen, the first member of a specific binding pair is an antibody specifically reactive with the antigen, the second member of a specific binding pair is an antibody specifically reactive with the antigen, and the third member of a specific binding pair is the antigen.

Claim 28

An assay of claim 17 wherein the analyte of interest is an antigen, the first member of a specific binding pair is an antibody specifically reactive with the antigen, the second member of a specific binding pair is an antibody specifically reactive with the antigen and the third member of a specific binding pair is an antigen specifically reactive with the first member of a specific binding pair and the second member of a specific binding pair.