Multiple hybrid immunoassay
OwnerJohn Lewis Emerson Campbell, Pamela Anne Frank +4inventors
InventorsJohn Lewis Emerson Campbell, Pamela Anne Frank, Meghan Elizabeth Hawkes, Shannon Reishma Lobin, Cary James Miller, Zhen Yang
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Abstract
The invention relates to compositions and methods for the immunoassay of an analyte of interest. The analyte is detected in an immunoassay using three or more antibodies, wherein each antibody specifically binds to a different epitope on the analyte. When the analyte of interest in a clinical marker for an acute disease, the detection of the analyte by immunoassay is a diagnosis of the occurrence of the disease.
Claims41
An immunoassay composition for detecting an analyte comprising at least three different antibodies, the at least three different antibodies being capable of binding to at least three different epitopes on an analyte, in which at least two of the at least three different antibodies are capable of binding to at least two different epitopes on a subform of the analyte, and in which at least one of the at least three different epitopes on the analyte is unavailable for binding on the subform.
An immunoassay composition for detecting an analyte comprising m different antibodies,
- (a) in which at least n of the m different antibodies are capable of binding to n different epitopes on an analyte, and
- (b) in which no more than n−1 of the n different epitopes on the analyte are available for binding on a subform of the analyte,
- provided that m and n are greater than or equal to 3, and m is greater than or equal to n.
An immunoassay composition for detecting an analyte comprising four different antibodies, the four different antibodies being capable of binding to four different epitopes on an analyte, in which at least one of four different epitopes on the analyte is unavailable for binding on a subform of the analyte, in which a first and second of the four different antibodies are conjugated to a surface, in which a third and fourth of the four different antibodies are each conjugated to an enzyme, in which at least one of the first and second antibodies is capable of binding to at least one epitope on the subform of the analyte, and in which at least one of the third and fourth antibodies is capable of binding to at least one epitope on the subform.
An immunoassay composition for detecting an analyte comprising m different antibodies,
- (a) in which n of the m different antibodies are capable of binding to n different epitopes on an analyte,
- (b) in which no more than n−1 of the n different epitopes on the analyte are available for binding on a subform of the analyte,
- (c) in which p of the n different antibodies are conjugated to a surface,
- (d) in which p of the n different antibodies are each conjugated to an enzyme,
- (e) in which at least p−1 of the p antibodies in step (c) is capable of binding to at least p−1 different epitopes on the subform of the analyte, and
- (f) in which at least p−1 of the p antibodies in step (d), is capable of binding to at least p−1 different epitopes on the subform of the analyte.
- provided that m and n are 4, and that p is 2.
An immunoassay device comprising one or more sensing elements and at least one surface on which an immunoassay can be conducted for detecting an analyte, which at least one surface comprises at least two different antibodies, the at least two different antibodies being capable of binding to at least two different epitopes on the analyte, in which at least one of the at least two different antibodies is capable of binding to at least one epitope on a subform of the analyte, in which at least one of the at least two different epitopes on the analyte is unavailable for binding on the subform, and in which the at least two different antibodies are bound to the at least one surface.
An immunoassay device comprising one or more sensing elements and at least one surface on which an immunoassay can be conducted for detecting an analyte,
- (a) in which at least one surface comprises m different antibodies,
- (b) in which at least n of the m different antibodies are capable of binding to n different epitopes on the analyte
- (c) in which no more than n−1 of the n different epitopes on the analyte are available for binding on a subform of the analyte, and
- (d) in which at least n of the m different antibodies are bound to the at least one surface,
- provided that m and n are greater than or equal to 2, and m is greater than or equal to n.
An immunoassay kit comprising in a suitable container, at least three different antibodies, the at least three antibodies being capable of binding to at least three different epitopes on an analyte, in which at least two of the at least three different antibodies are capable of binding to at least two different epitopes on a subform of the analyte, and in which at least one of the at least three different epitopes on the analyte is unavailable for binding on the subform.
An immunoassay kit comprising in a suitable container m different antibodies,
- (a) in which at least n of the m different antibodies are capable of binding to n different epitopes on an analyte, and
- (b) in which no more than n−1 of the n different epitopes on the analyte are available for binding on a subform of the analyte,
- provided that m and n are greater than or equal to 3, and m is greater than or equal to n.
A sandwich immunoassay product comprising an analyte and a subform of the analyte, in which at least three different epitopes on the analyte are available for binding by at least three different antibodies, in which at least two of the three different antibodies are bound to a different epitope on the analyte, in which two of the three different epitopes on the analyte are available for binding on the subform, in which at least two of the three different antibodies are bound to a different epitope on the subform, and in which at least one of the at least three different epitopes on the analyte is unavailable for binding on the subform.
A sandwich immunoassay product comprising an analyte, a subform of the analyte, and at least 2 of m different antibodies,
- (a) in which the analyte includes at least n different epitopes each capable of being bound by one of the m different antibodies,
- (b) in which at least 2 of the m different antibodies are each bound to a different epitope on the analyte,
- (c) in which no more than n−1 of the n different epitopes on the analyte are available for binding on a subform of the analyte, and
- (d) in which at least 2 of the m different antibodies are each bound to a different epitope on the subform of the analyte,
- provided that m and n are greater than or equal to 3, and that m can be greater than or equal to n.
A method of determining whether a patient has suffered a myocardial infarction comprising:
- (a) applying a sample from a patient suspected of suffering a myocardial infarction to a surface to which is bound at least two antibodies, in which the at least two antibodies are capable of binding to at least two different epitopes on cTnI, in which at least one of the at least two antibodies is capable of binding to at least one different epitope on a subform of cTnI, and in which at least one epitope of cTnI is unavailable for binding on the subform;
- (b) adding a reagent comprising a third antibody which binds to yet another epitope on cTnI and the subform;
- and
- (c) determining the extent of binding of the third antibody.
A method of determining whether a patient has suffered a myocardial infarction comprising:
- (a) applying a sample from a patient suspected of suffering a myocardial infarction to a surface to which is bound a first antibody which is capable of binding to a first epitope on cTnI and a subform of cTnI;
- (b) adding separately or together at least a second and third antibody, in which the at least second and third antibodies are capable of binding to at least two different epitopes on cTnI, in which the at least second and third antibodies are not capable of binding to the first epitope, in which at least one of the at least second and third antibodies are each capable of binding to at least one different epitope on the subform of cTnI, and in which at least one epitope of cTnI is unavailable for binding on the subform;
- and
- (c) determining the extent of binding of the at least second and third antibodies.
Dependencies are inferred from the claim text.