Method for immunochromatographic analysis

Claim 1

A method for conducting an assay for an analyte, which comprises: causing a specific binding pair (sbp) member in a first aqueous medium suspected of containing an analyte to become bound to a specific binding partner non-diffusively bound to at least a portion of a first bibulous member by contacting a portion of said first bibulous member with said medium, said first bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said first bibulous member and at least a portion of said absorbent member by capillary action; causing said first bibulous member to come into liquid receiving relationship with a second bibulous member; causing a reagent, which is capable of binding to said sbp member or a binding partner thereto, in a second aqueous medium to be specifically bound by a specific binding partner non-diffusively bound on said second bibulous member in relation to the presence of said analyte in said first medium by contacting a portion of said first bibulous member with said second aqueous medium, under conditions wherein said second aqueous medium traverses said first bibulous member by capillary action and at least a portion of said second bibulous member; and determining the presence of said reagent on at least a portion of said second bibulous member, the presence of said reagent indicating the presence of said analyte.

Claim 2

A method for conducting an assay for an analyte, which comprises: causing a specific binding pair (sbp) member in a first aqueous medium suspected of containing an analyte to become bound to a specific binding partner non-diffusively bound to at least a portion of a first bibulous member by contacting a portion of said first bibulous member with said medium, said first bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said first bibulous member and at least a portion of said absorbent member by capillary action; causing said first bibulous member to come into liquid receiving relationship with a second bibulous member; causing a reagent, which is capable of binding to said sbp member or a binding partner thereto, in a second aqueous medium to be specifically bound by a specific binding partner non-diffusively bound on said second bibulous member in relation to the presence of said analyte in said first medium by contacting a portion of said first bibulous member with said second aqueous medium, under conditions wherein said second aqueous medium traverses said first bibulous member by capillary action and at least a portion of said second bibulous member; and determining the presence of said reagent on at least a portion of said second bibulous member, the presence of said reagent indicating the presence of said analyte.

Claim 7

A method for conducting an assay for digoxin, which comprises: contacting an end portion of a bibulous strip having a zone distal from said end portion, said zone having an antibody (Ab 1 ) for an organic molecule of molecular weight less than 1500 non-diffusively bound thereto with a first aqueous medium containing a sample suspected of containing digoxin, wherein a conjugate comprising an antibody for digoxin bound to said organic molecule is present in said first medium or diffusively bound on said bibulous strip, under conditions wherein said first medium traverses said bibulous strip and at least a portion of said absorbent member by capillary action and said conjugate becomes non-diffusively bound to said Ab 1 and digoxin, if present, becomes bound to said conjugate; terminating the liquid receiving relationship between said bibulous strip and said absorbent member; providing in combination in a second aqueous medium said end portion of said bibulous strip and a reagent comprising a digoxin analog bound to an enzyme, said bibulous strip being in liquid receiving relationship with an immunochromatographic strip having non-diffusively bound thereto an antibody for digoxin, under conditions wherein said second medium traverses said bibulous strip and at least a portion of said immunochromatographic strip by capillary action, with the proviso that said bibulous strip is in liquid receiving relationship with said absorbent member and said immunochromatographic strip at substantially different times; and permitting a signal producing system including said enzyme to produce a signal defining a border on said immunochromatographic strip, the position of said border being related to the amount of analyte in said sample.

Claim 8

A method for conducting an assay for digoxin, which comprises: contacting an end portion of a bibulous strip having a zone distal from said end portion, said zone having an antibody (Ab 1 ) for an organic molecule of molecular weight less than 1500 non-diffusively bound thereto with a first aqueous medium containing a sample suspected of containing digoxin, wherein a conjugate comprising an antibody for digoxin bound to said organic molecule is present in said first medium or diffusively bound on said bibulous strip, under conditions wherein said first medium traverses said bibulous strip and at least a portion of said absorbent member by capillary action and said conjugate becomes non-diffusively bound to said Ab 1 and digoxin, if present, becomes bound to said conjugate; terminating the liquid receiving relationship between said bibulous strip and said absorbent member; providing in combination in a second aqueous medium said end portion of said bibulous strip and a reagent comprising a digoxin analog bound to an enzyme, said bibulous strip being in liquid receiving relationship with an immunochromatographic strip having non-diffusively bound thereto an antibody for digoxin, under conditions wherein said second medium traverses said bibulous strip and at least a portion of said immunochromatographic strip by capillary action, with the proviso that said bibulous strip is in liquid receiving relationship with said absorbent member and said immunochromatographic strip at substantially different times; and permitting a signal producing system including said enzyme to produce a signal defining a border on said immunochromatographic strip, the position of said border being related to the amount of analyte in said sample.

Claim 9

A method for conducting an assay for digoxin, said method comprising: contacting an end portion of a bibulous strip to which is non-diffusively bound a digoxin analog with a first aqueous medium containing a sample suspected of containing digoxin, wherein an antibody for digoxin (Ab D ) is present in said first medium or diffusively bound on said bibulous strip, said bibulous strip being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous strip and at least a portion of said absorbent member by capillary action and said first antibody becomes non-diffusively bound to said digoxin analog when digoxin is not present in said sample; terminating the liquid receiving relationship between said bibulous strip and said absorbent member; providing in combination in a second aqueous medium said end portion of said bibulous strip and a conjugate comprising an antibody for said Ab D bound to an enzyme, said bibulous strip being in liquid receiving relationship with an immunochromatographic strip having non-diffusively bound thereto a receptor for said antibody for Ab D , under conditions wherein said second medium traverses said bibulous strip is in liquid receiving relationship with said absorbent member and said immunochromatographic strip at substantially different times; and permitting a signal producing system including said enzyme to produce a signal defining a border on said immunochromatographic strip, the position of said border being related to the amount of analyte in said first medium.

Claim 10

A method for conducting an assay for digoxin, said method comprising: contacting an end portion of a bibulous strip to which is non-diffusively bound a digoxin analog with a first aqueous medium containing a sample suspected of containing digoxin, wherein an antibody for digoxin (Ab D ) is present in said first medium or diffusively bound on said bibulous strip, said bibulous strip being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous strip and at least a portion of said absorbent member by capillary action and said first antibody becomes non-diffusively bound to said digoxin analog when digoxin is not present in said sample; terminating the liquid receiving relationship between said bibulous strip and said absorbent member; providing in combination in a second aqueous medium said end portion of said bibulous strip and a conjugate comprising an antibody for said Ab D bound to an enzyme, said bibulous strip being in liquid receiving relationship with an immunochromatographic strip having non-diffusively bound thereto a receptor for said antibody for Ab D , under conditions wherein said second medium traverses said bibulous strip is in liquid receiving relationship with said absorbent member and said immunochromatographic strip at substantially different times; and permitting a signal producing system including said enzyme to produce a signal defining a border on said immunochromatographic strip, the position of said border being related to the amount of analyte in said first medium.

Claim 11

A method for conducting an assay for an analyte, said method comprising: contacting an end portion of a bibulous strip to which is non-diffusively bound an antibody for digoxin with a first aqueous medium containing a sample suspected of containing digoxin, said bibulous strip being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous strip and at least a portion of said absorbent member by capillary action; terminating the liquid receiving relationship between said bibulous strip and said absorbent member; providing in combination in a second aqueous medium said end portion of said bibulous strip and a conjugate comprising a digoxin analog bound to an enzyme, said bibulous strip being in liquid receiving relationship with an immunochromatographic strip having non-diffusively bound thereto a receptor for conjugate, under conditions wherein said second medium traverses said bibulous strip and at least a portion of said immunochromatographic strip by capillary action, with the proviso that said bibulous strip is in liquid receiving relationship with said absorbent member and said immunochromatographic strip at substantially different times; and permitting a signal producing system including said enzyme to produce a signal defining a border on said immunochromatographic strip, the position of said border being related to the amount of analyte in said first medium.

Claim 12

A method for conducting an assay for an analyte, said method comprising: contacting an end portion of a bibulous strip to which is non-diffusively bound an antibody for digoxin with a first aqueous medium containing a sample suspected of containing digoxin, said bibulous strip being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous strip and at least a portion of said absorbent member by capillary action; terminating the liquid receiving relationship between said bibulous strip and said absorbent member; providing in combination in a second aqueous medium said end portion of said bibulous strip and a conjugate comprising a digoxin analog bound to an enzyme, said bibulous strip being in liquid receiving relationship with an immunochromatographic strip having non-diffusively bound thereto a receptor for conjugate, under conditions wherein said second medium traverses said bibulous strip and at least a portion of said immunochromatographic strip by capillary action, with the proviso that said bibulous strip is in liquid receiving relationship with said absorbent member and said immunochromatographic strip at substantially different times; and permitting a signal producing system including said enzyme to produce a signal defining a border on said immunochromatographic strip, the position of said border being related to the amount of analyte in said first medium.

Claim 13

A method for conducting an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a first bibulous member, at least a portion thereof having a specific binding pair (sbp) member non-diffusively bound thereto and (2) a sample suspected of containing an analyte with the proviso that, when said analyte is not capable of binding to said sbp member, a first reagent capable of binding to said analyte and to said sbp member is present in said medium or diffusively bound on said first bibulous member, said first bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said first bibulous member and at least a portion of said absorbent member by capillary action; bringing said first bibulous member into liquid receiving relationship with a second bibulous member; providing in combination in a second aqueous medium a portion of said first bibulous member and a second reagent, said second reagent being capable of binding to said sbp member or said first reagent in relation to the presence of said analyte in said first medium, said second bibulous member having non-diffusively bound to at least a portion thereof a specific binding partner for said second reagent, under conditions wherein said second medium traverses at least a portion of said first bibulous member and at least a portion of said second bibulous member by capillary action; and determining the presence said second reagent bound to second bibulous member, the presence thereof being related to the amount of said analyte in said sample.

Claim 14

A method for conducting an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a first bibulous member, at least a portion thereof having a specific binding pair (sbp) member non-diffusively bound thereto and (2) a sample suspected of containing an analyte with the proviso that, when said analyte is not capable of binding to said sbp member, a first reagent capable of binding to said analyte and to said sbp member is present in said medium or diffusively bound on said first bibulous member, said first bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said first bibulous member and at least a portion of said absorbent member by capillary action; bringing said first bibulous member into liquid receiving relationship with a second bibulous member; providing in combination in a second aqueous medium a portion of said first bibulous member and a second reagent, said second reagent being capable of binding to said sbp member or said first reagent in relation to the presence of said analyte in said first medium, said second bibulous member having non-diffusively bound to at least a portion thereof a specific binding partner for said second reagent, under conditions wherein said second medium traverses at least a portion of said first bibulous member and at least a portion of said second bibulous member by capillary action; and determining the presence said second reagent bound to second bibulous member, the presence thereof being related to the amount of said analyte in said sample.

Claim 37

A method for conducting an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a bibulous member to at least a portion of which is non-diffusively bound a first specific binding pair (sbp) member and (2) a sample suspected of containing an analyte, wherein a conjugate comprising a second sbp member capable of binding said analyte and a third sbp member complementary to said first sbp member is present in said medium or diffusively bound on said bibulous member, said bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous member and at least a portion of said absorbent member by capillary action and said conjugate becomes non-diffusively bound to said bibulous member; terminating the liquid receiving relationship between said bibulous member and said absorbent member; providing in combination in a second aqueous medium a portion of said bibulous member and a reagent comprising an analyte analog bound to, or capable of becoming bound to, a member of a signal producing system, said bibulous member being in liquid receiving relationship with an immunochromatographic member having non-diffusively bound thereto a binding partner for said reagent, under conditions wherein said second medium traverses said bibulous member and at least a portion of said immunochromatographic member by capillary action, with the proviso that said bibulous member is in liquid receiving relationship with said absorbent member and said immunochromatographic member at substantially different times; and permitting said signal producing system to produce a signal defining a border on said immunochromatographic member, the position of said border being related to the amount of analyte in said first medium.

Claim 38

A method for conducting an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a bibulous member to at least a portion of which is non-diffusively bound a first specific binding pair (sbp) member and (2) a sample suspected of containing an analyte, wherein a conjugate comprising a second sbp member capable of binding said analyte and a third sbp member complementary to said first sbp member is present in said medium or diffusively bound on said bibulous member, said bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous member and at least a portion of said absorbent member by capillary action and said conjugate becomes non-diffusively bound to said bibulous member; terminating the liquid receiving relationship between said bibulous member and said absorbent member; providing in combination in a second aqueous medium a portion of said bibulous member and a reagent comprising an analyte analog bound to, or capable of becoming bound to, a member of a signal producing system, said bibulous member being in liquid receiving relationship with an immunochromatographic member having non-diffusively bound thereto a binding partner for said reagent, under conditions wherein said second medium traverses said bibulous member and at least a portion of said immunochromatographic member by capillary action, with the proviso that said bibulous member is in liquid receiving relationship with said absorbent member and said immunochromatographic member at substantially different times; and permitting said signal producing system to produce a signal defining a border on said immunochromatographic member, the position of said border being related to the amount of analyte in said first medium.

Claim 53

A method for conducting an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a bibulous member to which is non-diffusively bound an analyte analog and (2) a sample suspected of containing an analyte, wherein a first reagent capable of binding said analyte is present in said medium or diffusively bound on said bibulous member, said bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous member and at least a portion of said absorbent member by capillary action and said first reagent becomes non-diffusively bound to said bibulous member when said analyte is not present in said first medium; terminating the liquid receiving relationship between said bibulous member and said absorbent member; providing in combination in a second aqueous medium a portion of said bibulous member and a second reagent, said second reagent being bound to, or capable of becoming bound to, a member of a signal producing system and being capable of binding to said first reagent when said first reagent is diffusively bound to said bibulous member, said bibulous member being in liquid receiving relationship with an immunochromatographic member having non-diffusively bound thereto a specific binding partner for said second reagent, under conditions under which said second medium traverses said bibulous member and at least a portion of said immunochromatographic member by capillary action, with the proviso that said bibulous member is in liquid receiving relationship with said absorbent member and said immunochromatographic member at substantially different times; and permitting said signal producing system to produce a signal defining a border on said immunochromatographic member, the position of said border being related to the amount of analyte in said first medium.

Claim 54

A method for conducting an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a bibulous member to which is non-diffusively bound an analyte analog and (2) a sample suspected of containing an analyte, wherein a first reagent capable of binding said analyte is present in said medium or diffusively bound on said bibulous member, said bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous member and at least a portion of said absorbent member by capillary action and said first reagent becomes non-diffusively bound to said bibulous member when said analyte is not present in said first medium; terminating the liquid receiving relationship between said bibulous member and said absorbent member; providing in combination in a second aqueous medium a portion of said bibulous member and a second reagent, said second reagent being bound to, or capable of becoming bound to, a member of a signal producing system and being capable of binding to said first reagent when said first reagent is diffusively bound to said bibulous member, said bibulous member being in liquid receiving relationship with an immunochromatographic member having non-diffusively bound thereto a specific binding partner for said second reagent, under conditions under which said second medium traverses said bibulous member and at least a portion of said immunochromatographic member by capillary action, with the proviso that said bibulous member is in liquid receiving relationship with said absorbent member and said immunochromatographic member at substantially different times; and permitting said signal producing system to produce a signal defining a border on said immunochromatographic member, the position of said border being related to the amount of analyte in said first medium.

Claim 66

A method for conductinq an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a bibulous member to which is non-diffusively bound an antibody for said analyte and (2) a sample suspected of containing an analyte, said bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous member and at least a portion of said absorbent member by capillary action; terminating the liquid receiving relationship between said bibulous member and said absorbent member; providing in combination in a second aqueous medium a portion of said bibulous member and a reagent, said reagent being bound to, or capable of becoming bound to, a member of a signal producing system and being capable of binding to said antibody for said analyte, said bibulous member being in liquid receiving relationship with an immunochromatographic member having non-diffusively bound thereto a specific binding partner for said reagent, under conditions under which said second medium traverses said bibulous member and at least a portion of said immunochromatographic member by capillary action, with the proviso that said bibulous member is in liquid receiving relationship with said absorbent member and said immunochromatographic member at substantially different times; and permitting said signal producing system to produce a signal defining a border on said immunochromatographic member, the position of said border being related to the amount of analyte in said first medium.

Claim 67

A method for conductinq an assay for an analyte, said method comprising: providing in combination in a first aqueous medium (1) a portion of a bibulous member to which is non-diffusively bound an antibody for said analyte and (2) a sample suspected of containing an analyte, said bibulous member being in liquid receiving relationship with an absorbent member, under conditions wherein said first medium traverses said bibulous member and at least a portion of said absorbent member by capillary action; terminating the liquid receiving relationship between said bibulous member and said absorbent member; providing in combination in a second aqueous medium a portion of said bibulous member and a reagent, said reagent being bound to, or capable of becoming bound to, a member of a signal producing system and being capable of binding to said antibody for said analyte, said bibulous member being in liquid receiving relationship with an immunochromatographic member having non-diffusively bound thereto a specific binding partner for said reagent, under conditions under which said second medium traverses said bibulous member and at least a portion of said immunochromatographic member by capillary action, with the proviso that said bibulous member is in liquid receiving relationship with said absorbent member and said immunochromatographic member at substantially different times; and permitting said signal producing system to produce a signal defining a border on said immunochromatographic member, the position of said border being related to the amount of analyte in said first medium.