Products for protein analyte detection assays using antibodies

A product for detecting a plurality of protein analytes in a sample, comprising:

• (i) a plurality of proximity probe pairs, wherein each proximity probe pair comprises a first proximity probe and a second proximity probe, and each proximity probe comprises:
• (a) an antibody or antibody fragment specific for a protein analyte;
• and
• (b) a nucleic acid domain,
• wherein both proximity probes within each proximity probe pair comprise antibodies or antibody fragments specific for the same protein analyte, and can simultaneously bind to the protein analyte, wherein all proximity probe pairs specific for the same protein analyte comprise a species of proximity probe pairs, and each species of proximity probe pairs is specific for a different protein analyte;
• wherein the nucleic acid domain of each proximity probe comprises an identification (ID) sequence and at least a first hybridisation sequence, wherein the ID sequences of each species of proximity probe pairs correspond to a particular protein analyte;
• and
• wherein in each proximity probe pair, the first proximity probe and the second proximity probe comprise paired hybridisation sequences and one or both nucleic acid domains are partially double-stranded,
• wherein at least one partially double-stranded nucleic acid domain comprises
• a first oligonucleotide conjugated to the analyte-binding domain;
• and
• a hybridisation oligonucleotide comprising, from 5′ to 3′, the first hybridisation sequence, the ID sequence and a second hybridisation sequence, and the ID sequence is located in a single-stranded part of the nucleic acid domain;
• (ii) for each species of proximity probe pairs, a splint oligonucleotide which comprises hybridisation sequences complementary to each of the paired hybridisation sequences of the first proximity probe and the second proximity probe;
• wherein the hybridisation sequences of each proximity probe pair are configured such that when the first proximity probe and the second proximity probe are bound to their protein analyte, the respective paired hybridisation sequences of the first proximity probe and the second proximity probe hybridise to the splint oligonucleotide;
• and wherein at least one pair of hybridisation sequences is shared by at least two species of proximity probe pairs;
• and
• (iii) a plurality of sample index oligonucleotides having nucleotide sequences for identifying respective source samples.

A product for detecting a plurality of protein analytes in a sample, comprising:

• (i) a plurality of proximity probe pairs, wherein each proximity probe pair comprises a first proximity probe and a second proximity probe, and each proximity probe comprises:
• (a) an antibody or antibody fragment specific for a protein analyte;
• and
• (b) a nucleic acid domain,
• wherein both proximity probes within each proximity probe pair comprise antibodies or antibody fragments specific for the same protein analyte, and can simultaneously bind to the protein analyte, wherein all proximity probe pairs specific for the same protein analyte comprise a species of proximity probe pairs, and each species of proximity probe pairs is specific for a different protein analyte;
• wherein the nucleic acid domain of each proximity probe comprises an identification (ID) sequence and at least a first hybridisation sequence, wherein the ID sequences of each species of proximity probe pairs correspond to a particular protein analyte;
• and
• wherein in each proximity probe pair, the first proximity probe and the second proximity probe comprise paired hybridisation sequences, one or both nucleic acid domains are partially double-stranded, and the nucleic acid domains of both proximity probes in a pair are conjugated to the protein-binding domain by their 5′ ends;
• (ii) for each species of proximity probe pairs, a splint oligonucleotide which comprises hybridisation sequences complementary to each of the paired hybridisation sequences of the first proximity probe and the second proximity probe;
• wherein the hybridisation sequences of each proximity probe pair are configured such that when the first proximity probe and the second proximity probe are bound to their protein analyte, the respective paired hybridisation sequences of the first proximity probe and the second proximity probe hybridise to the splint oligonucleotide;
• and wherein at least one pair of hybridisation sequences is shared by at least two species of proximity probe pairs;
• and
• (iii) a plurality of sample index oligonucleotides having nucleotide sequences for identifying respective source samples.