Method of detecting autoantibodies against fc epsilon receptor i

Abstract

The present disclosure relates to methods of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand's receptor, and uses of the method thereof.

Description

[0001] METHODS OF DETECTING A TARGET LIGAND CAPABLE OF INDUCING RECEPTOR OLIGOMERIZATION

[0002] RELATED APPLICATION DATA

[0003] The present application claims priority from United States Patent Application No. 63 / 702,41 filed 2 October 2024 entitled “Methods of detecting a target ligand capable of inducing receptor oligomerization”, the entire contents of which is hereby incorporated by reference.

[0004] FIELD

[0005] The present disclosure relates to methods of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand’s receptor, and uses of the method thereof.

[0006] BACKGROUND

[0007] Many types of cell surface receptors undergo dimerization, oligomerization, or crosslinking following target ligand binding and / or interaction. These target ligand-receptor complexes then trigger a cascade of events that culminate in a cellular response, such as proliferation or altered decreased gene expression. This oligomerization serves as a surrogate for receptor function.

[0008] Whilst these target ligand-receptor complexes are essential for maintaining normal physiological processes, they also play a critical role in disease, and adverse drug reactions.

[0009] Existing methods for identification of unwanted target ligands (or biomarkers) in the blood involve immunoassays such as enzyme-linked immunosorbent assay (ELISA) and fluorescence linked immunosorbent assay (FLISA). In these standard assays, the target analyte within a sample is brought into direct contact with the immobilized receptor (i.e., antigen) and forms a direct bond with its target if present in the sample. The immobilized antigen is generally labelled to allow target identification. However, lack of signal due to low target concentrations or high background signal are common problems with existing immunoassay techniques.

[0010] More recently, the Nucleic Acid-Linked Immuno- Sandwich Assay (NULISA™) has provided increased sensitivity of traditional proximity ligation assays through a dual capture and release mechanism incorporated into oligonucleotide-conjugated antibodies for detection of the target analyte. Whilst the NULISA™ assay allows for detection and quantification of target biomolecules, the assay does not provide any indication regarding function of the receptor-analyte interaction. For example, whether or not the target analyte will initiate a cellular response as a result of receptor oligomerization.

[0011] Accordingly, it will be apparent to the skilled person that there is a need in the art for methods or assays for detection of target ligands in a sample, for example, target ligands that are present at low levels in a sample and / or that are capable of inducing receptor oligomerization and e.g., unwanted cellular responses. SUMMARY

[0012] The present disclosure is based on the inventors’ development of a method of detecting a target ligand in a sample, wherein the ligand is capable of inducing oligomerization and / or crosslinking of the ligand’s receptor. In particular, the inventors proceeded against conventional wisdom of existing proximity assays and used the biomarker (i.e., FcsRI receptor) to detect the ligand (i.e., anti-FcsRI antibodies) against the biomarker (as opposed to using the ligand to detect the biomarker). In particular, the inventors found that use of nucleic acid linked FcsRI receptors in a proximity assay resulted in detection of the target anti-FcsRI antibodies with attomolar sensitivity. The inventors also found that using two FcsRI receptor a-chains to detect the target ligand worked surprisingly better than one FcsRI receptor a-chain and one FcsRI receptor P-chain. Surprisingly, it was also found that the assay served as a surrogate for in vitro assays for FcsRI receptor oligomerization and function, including mast cell activation and basophil degranulation, enabling clear differentiation between normal samples and samples comprising a level of target ligand capable of inducing unwanted cellular responses. In particular, the assay by the inventors is particularly useful in detecting low levels of target ligands with high specificity and sensitivity.

[0013] Accordingly, the findings by the inventors provide the basis for methods of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand’s receptor.

[0014] The present disclosure provides a method of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand’s receptor, the method comprising:

[0015] (i) contacting the sample comprising the target ligand to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the target ligand in the sample to thereby form an immunocomplex;

[0016] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0017] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the target ligand in the sample.

[0018] In one example, prior to contacting the immunocomplex to the first capture probe immobilized on the first solid matrix, the method comprises:

[0019] (i) contacting the immunocomplex to a second capture probe immobilized on a second solid matrix, wherein the second capture probe binds the second nucleic acid tag;

[0020] (ii) washing the second solid matrix to remove unbound immunocomplex; and

[0021] (iii) releasing the bound immunocomplex from the second immobilized capture probe.

[0022] The present disclosure provides a method of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand’s receptor, the method comprising:

[0023] (i) contacting the sample comprising the target ligand to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the target ligand in the sample to thereby form an immunocomplex;

[0024] (ii) contacting the immunocomplex to a second capture probe immobilized on a second solid matrix, wherein the second capture probe binds the second nucleic acid tag;

[0025] (iii)washing the second solid matrix to remove unbound immunocomplex;

[0026] (iv)releasing the bound immunocomplex from the second immobilized capture probe;

[0027] (v) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0028] (vi)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0029] (vii) detecting the reporter, wherein the presence of the reporter is indicative of the presence of the target ligand in the sample.

[0030] In one example, the nucleic acid reporter is generated by:

[0031] (i) linking the first nucleic acid tag and the second nucleic acid tag, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid tag; or (ii) linking the first nucleic acid tag and the second nucleic acid label, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid label; or

[0032] (iii)linking the first nucleic acid label and the second nucleic acid tag, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid tag; or

[0033] (iv)linking the first nucleic acid label and the second nucleic acid label and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid label.

[0034] In one example, the nucleic acid reporter is generated by linking the first nucleic acid tag and the second nucleic acid tag and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid tag

[0035] In one example, the nucleic acid reporter is generated by linking the first nucleic acid tag and the second nucleic acid label and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid label.

[0036] In one example, the nucleic acid reporter is generated by linking the first nucleic acid label and the second nucleic acid tag and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid tag.

[0037] In one example, the nucleic acid reporter is generated by linking the first nucleic acid label and the second nucleic acid label and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid label.

[0038] In one example, the nucleic acid reporter is formed by proximity ligation or proximity extension. In another example, the nucleic acid reporter is formed by proximity ligation. In a further example, the nucleic acid reporter is formed by proximity extension.

[0039] In one example, the method further comprises washing the first solid matrix to remove unbound immunocomplex.

[0040] In one example, the first capture probe is:

[0041] (i) a protein that specifically binds to the first nucleic acid tag;

[0042] (ii) a protein and nucleic acid complex that specifically binds to the first nucleic acid tag;

[0043] (iii) a nucleic acid molecule, wherein the first capture probe or a fragment thereof is complementary to the first nucleic acid tag or a fragment thereof; or

[0044] (iv) a nucleic acid molecule, wherein the first capture probe or a fragment thereof hybridizes with the first nucleic acid tag or a fragment thereof.

[0045] In one example, the first capture probe is a protein that specifically binds to the first nucleic acid tag.

[0046] In one example, the first capture probe is a protein and nucleic acid complex that specifically binds to the first nucleic acid tag.

[0047] In one example, the first capture probe is a nucleic acid molecule, wherein the first capture probe or a fragment thereof is complementary to the first nucleic acid tag or a fragment thereof. In one example, the first capture probe is a nucleic acid molecule, wherein the first capture probe or a fragment thereof hybridizes with the first nucleic acid tag or a fragment thereof.

[0048] In one example, the first capture probe comprises a biotin binding protein. For example, the biotin binding protein is streptavidin, avidin or neutravidin. In one example, the biotin binding protein is streptavidin. In another example, the biotin binding protein is avidin. In a further example, the biotin binding protein is neutravidin.

[0049] In one example, the biotinylated tail sequence of the first nucleic acid tag binds to the biotin binding protein. For example, the biotinylated tail sequence of the first nucleic acid tag binds to the biotin binding protein of the first capture probe. In one example, the biotinylated tail sequence of the first nucleic acid tag binds to the streptavidin, avidin or neutravidin of the first capture probe. In another example, the biotinylated tail sequence of the first nucleic acid tag binds to the streptavidin of the first capture probe. In a further example, the biotinylated tail sequence of the first nucleic acid tag binds to the avidin of the first capture probe. In one example, the biotinylated tail sequence of the first nucleic acid tag binds to the neutravidin of the first capture probe.

[0050] In one example, the first nucleic acid tag binds to the first capture probe via a thioester group, a disulfide linkage, a cleavable linkage or a protein-protein interaction. In one example, the first nucleic acid tag binds to the first capture probe via a thioester group. In another example, the first nucleic acid tag binds to the first capture probe via a disulfide linkage. In a further example, the first nucleic acid tag binds to the first capture probe by a cleavable linkage. In one example, the first nucleic acid tag binds to the first capture probe by a protein-protein interaction.

[0051] In one example, the second capture probe is:

[0052] (i) a protein that specifically binds to the second nucleic acid tag;

[0053] (ii) a protein and nucleic acid complex that specifically binds to the second nucleic acid tag;

[0054] (iii) a nucleic acid molecule, wherein the second capture probe or a fragment thereof is complementary to the second nucleic acid tag or a fragment thereof; or

[0055] (iv) a nucleic acid molecule, wherein the second capture probe or a fragment thereof hybridizes with the second nucleic acid tag or a fragment thereof.

[0056] In one example, the second capture probe is a protein that specifically binds to the second nucleic acid tag.

[0057] In one example, the second capture probe is a protein and nucleic acid complex that specifically binds to the second nucleic acid tag.

[0058] In one example, the second capture probe is a nucleic acid molecule, wherein the second capture probe or a fragment thereof is complementary to the second nucleic acid tag or a fragment thereof.

[0059] In one example, the second capture probe is a nucleic acid molecule, wherein the second capture probe or a fragment thereof hybridizes with the second nucleic acid tag or a fragment thereof.

[0060] In one example, the second capture probe comprises a T, an A, a C and / or a G rich sequence complementary to the A, a T, a G and / or a C rich tail sequence of the second nucleic acid tag. In one example, the second capture probe comprises a T rich sequence complementary to the A rich tail sequence of the second nucleic acid tag. In another example, the second capture probe comprises an A rich sequence complementary to the T rich sequence of the second nucleic acid tag. In a further example, the second capture probe comprises a C rich sequence complementary to the G rich tail sequence of the second nucleic acid tag. In one example, the second capture probe comprises a G rich sequence complementary to the C rich tail sequence of the second nucleic acid tag.

[0061] In one example, the A, T, G and / or C rich tail sequence of the second nucleic acid tag binds the complementary T, A, C and / or G rich sequence of the second capture probe. In one example, the A rich tail sequence of the second nucleic acid tag binds to the complementary T rich sequence of the second capture probe. In another example, the T rich tail sequence of the second nucleic acid tag binds to the complementary A rich sequence of the second capture probe. In a further example, the G rich tail sequence of the second nucleic acid tag binds to the complementary C rich sequence of the second capture probe. In one example, the C rich tail sequence of the second nucleic acid tag binds to the complementary G rich sequence of the second capture probe.

[0062] In one example, the second nucleic acid tag binds to the second capture probe via a thioester group, a disulfide linkage, a cleavable linkage or a protein-protein interaction. In one example, the second nucleic acid tag binds to the second capture probe via a thioester group. In another example, the second nucleic acid tag binds to the second capture probe via a disulfide linkage. In a further example, the second nucleic acid tag binds to the second capture probe by a cleavable linkage. In one example, the second nucleic acid tag binds to the second capture probe by a protein-protein interaction.

[0063] In one example,

[0064] (i) the first nucleic acid tag is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. fused to the first receptor, fragment or epitope thereof; c. conjugated to the first receptor, fragment or epitope thereof; d. non-covalently bound to the first receptor, fragment or epitope thereof; e. conjugated to the first nucleic acid label; f. non-covalently bound to the first nucleic acid label; or g. part of the first nucleic acid label; and / or

[0065] (ii) the second nucleic acid tag is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. fused to the second receptor, fragment or epitope thereof; c. conjugated to the second receptor, fragment or epitope thereof; d. non-covalently bound to the second receptor, fragment or epitope thereof; e. conjugated to the second nucleic acid label; f. non-covalently bound to the second nucleic acid label; or g. part of the second nucleic acid label.

[0066] In one example, the first nucleic acid tag is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. fused to the first receptor, fragment or epitope thereof; c. conjugated to the first receptor, fragment or epitope thereof; d. non-covalently bound to the first receptor, fragment or epitope thereof; e. conjugated to the first nucleic acid label; f. non-covalently bound to the first nucleic acid label; or g. part of the first nucleic acid label; and / or

[0067] In one example, the first nucleic acid tag is directly or indirectly bound to the first receptor, fragment, or epitope thereof.

[0068] In one example, the first nucleic acid tag is fused to the first receptor, fragment or epitope thereof.

[0069] In one example, the first nucleic acid tag is conjugated to the first receptor, fragment, or epitope thereof.

[0070] In one example, the first nucleic acid tag is non-covalently bound to the first receptor, fragment, or epitope thereof.

[0071] In one example, the first nucleic acid tag is conjugated to the first nucleic acid label.

[0072] In one example, the first nucleic acid tag is non-covalently bound to the first nucleic acid label.

[0073] In one example, the first nucleic acid tag is part of the first nucleic acid label.

[0074] In one example, the second nucleic acid tag is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. fused to the second receptor, fragment or epitope thereof; c. conjugated to the second receptor, fragment or epitope thereof; d. non-covalently bound to the second receptor, fragment or epitope thereof; e. conjugated to the second nucleic acid label; f. non-covalently bound to the second nucleic acid label; or g. part of the second nucleic acid label.

[0075] In one example, the second nucleic acid tag is directly or indirectly bound to the second receptor, fragment, or epitope thereof.

[0076] In one example, the second nucleic acid tag is fused to the second receptor, fragment, or epitope thereof.

[0077] In one example, the second nucleic acid tag is conjugated to the second receptor, fragment, or epitope.

[0078] In one example, the second nucleic acid tag is non-covalently bound to the second receptor, fragment, or epitope thereof.

[0079] In one example, the second nucleic acid tag is conjugated to the second nucleic acid label.

[0080] In one example, the second nucleic acid tag is non-covalently bound to the second nucleic acid label.

[0081] In one example, the second nucleic acid tag is part of the second nucleic acid label.

[0082] In one example,

[0083] (i) the first nucleic acid label is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. conjugated to the first receptor, fragment or epitope thereof; c. non-covalently bound to the first receptor, fragment or epitope thereof; d. conjugated to the first nucleic acid tag; e. non-covalently bound to the first nucleic acid tag; f. hybridizes with the first nucleic acid tag; or g. part of the first nucleic acid tag; and / or

[0084] (ii) the second nucleic acid label is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. conjugated to the second receptor, fragment or epitope thereof; c. non-covalently bound to the second receptor, fragment or epitope thereof; d. conjugated to the second nucleic acid tag; e. non-covalently bound to the second nucleic acid tag; f. hybridizes with the second nucleic acid tag; or g. part of the second nucleic acid tag.

[0085] In one example, the first nucleic acid label is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. conjugated to the first receptor, fragment or epitope thereof; c. non-covalently bound to the first receptor, fragment or epitope thereof; d. conjugated to the first nucleic acid tag; e. non-covalently bound to the first nucleic acid tag; f. hybridizes with the first nucleic acid tag; or g. part of the first nucleic acid tag; and / or

[0086] In one example, the first nucleic acid label is directly or indirectly bound to the first receptor, fragment, or epitope thereof.

[0087] In one example, the first nucleic acid label is conjugated to the first receptor, fragment, or epitope thereof.

[0088] In one example, the first nucleic acid label is non-covalently bound to the first receptor, fragment, or epitope thereof.

[0089] In one example, the first nucleic acid label is conjugated to the first nucleic acid tag.

[0090] In one example, the first nucleic acid label is non-covalently bound to the first nucleic acid tag.

[0091] In one example, the first nucleic acid label hybridizes with the first nucleic acid tag.

[0092] In one example, the first nucleic acid label is part of the first nucleic acid tag.

[0093] In one example, the second nucleic acid label is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. conjugated to the second receptor, fragment or epitope thereof; c. non-covalently bound to the second receptor, fragment or epitope thereof; d. conjugated to the second nucleic acid tag; e. non-covalently bound to the second nucleic acid tag; f. hybridizes with the second nucleic acid tag; or g. part of the second nucleic acid tag.

[0094] In one example, the second nucleic acid label is directly or indirectly bound to the second receptor, fragment, or epitope thereof.

[0095] In one example, the second nucleic acid label is conjugated to the second receptor, fragment, or epitope thereof.

[0096] In one example, the second nucleic acid label is non-covalently bound to the second receptor, fragment, or epitope thereof.

[0097] In one example, the second nucleic acid label is conjugated to the second nucleic acid tag.

[0098] In one example, the second nucleic acid label is non-covalently bound to the second nucleic acid tag.

[0099] In one example, the second nucleic acid label hybridizes with the second nucleic acid tag.

[0100] In one example, the second nucleic acid label is part of the second nucleic acid tag.

[0101] In one example:

[0102] (i) the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment or epitope thereof; and / or

[0103] (ii) the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment or epitope thereof.

[0104] In one example, the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment, or epitope thereof.

[0105] In one example, the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment, or epitope thereof.

[0106] In one example, the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment, or epitope thereof and the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment or epitope thereof.

[0107] In one example, the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment, or epitope thereof or the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment or epitope thereof.

[0108] In one example, the nucleic acid reporter comprises the first receptor barcode and / or second receptor barcode. In another example, the nucleic acid reporter comprises the first receptor barcode. In a further example, the nucleic acid reporter comprises the second receptor barcode. In one example, the nucleic acid reporter comprises the first receptor barcode and second receptor barcode. In another example, the nucleic acid reporter comprises the first receptor barcode or second receptor barcode

[0109] In one example:

[0110] (i) the first receptor barcode in the nucleic acid reporter is a complementary sequence of the first receptor barcode in the first nucleic acid label; and / or

[0111] (ii) the second receptor barcode in the nucleic acid reporter is a complementary sequence of the second receptor barcode in the second nucleic acid label. In one example, the first receptor barcode in the nucleic acid reporter is a complementary sequence of the first receptor barcode in the first nucleic acid label.

[0112] In one example, the second receptor barcode in the nucleic acid reporter is a complementary sequence of the second receptor barcode in the second nucleic acid label.

[0113] In one example, the first receptor barcode in the nucleic acid reporter is a complementary sequence of the first receptor barcode in the first nucleic acid label and the second receptor barcode in the nucleic acid reporter is a complementary sequence of the second receptor barcode in the second nucleic acid label.

[0114] In one example, the first receptor barcode in the nucleic acid reporter is a complementary sequence of the first receptor barcode in the first nucleic acid label or the second receptor barcode in the nucleic acid reporter is a complementary sequence of the second receptor barcode in the second nucleic acid label.

[0115] In one example, the first receptor barcode and the second receptor barcode are identical.

[0116] In one example, the first nucleic acid label and / or the second nucleic acid label further comprise a sample barcode. In another example, the first nucleic acid label further comprises a sample barcode. In a further example, the second nucleic acid label further comprises a sample barcode. In one example, the first nucleic acid label and the second nucleic acid label further comprise a sample barcode. In another example, the first nucleic acid label or the second nucleic acid label further comprise a sample barcode.

[0117] In one example, the nucleic acid reporter comprises the sample barcode.

[0118] In one example, the nucleic acid reporter comprises the first receptor barcode, the second receptor barcode and the sample barcode. In another example, the nucleic acid reporter comprises the first receptor barcode and the sample barcode. In a further example, the nucleic acid reporter comprises the second receptor barcode and the sample barcode. In one example, the nucleic acid reporter comprises the first receptor barcode or the second receptor barcode; and the sample barcode.

[0119] In one example, the method comprises detecting the nucleic acid reporter by polymerase chain reaction (PCR), next generation sequencing, Rolling Cycle Amplification (RCA), strand displacement amplification (SDA), Loop-Mediated Isothermal Amplification (LAMP), Recombinase Polymerase Amplification (RPA), a QuantiGene assay, a CRISPR-based detection technique, Nucleic Acid Sequence-Based Amplification (NASBA), Helicase-Dependent Amplification (HDA), fluorescence-based detection, or microarray techniques. In another example, the method comprises detecting the nucleic acid reporter by PCR. For example, the PCR is multiplexed quantitative PCR (qPCR) or multiplexed digital PCR. In one example, the PCR is qPCR. In another example, the PCR is multiplexed digital PCR. In a further example, the method comprises detecting the nucleic acid reporter by next generation sequencing. In one example, the method comprises detecting the nucleic acid reporter by RCA. In another example, the method comprises detecting the nucleic acid reporter by SDA. In a further example, the method comprises detecting the nucleic acid reporter by LAMP. In one example, the method comprises detecting the nucleic acid reporter by RPA. In another example, the method comprises detecting the nucleic acid reporter by QuantiGene assay. In one example, the method comprises detecting the nucleic acid reporter by CRISPR-based detection techniques. For example, Specific High-sensitivity Enzymatic Reporter unlocking (SHERLOCK) or DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR). In another example, the method comprises detecting the nucleic acid reporter by NASB A. In a further example, the method comprises detecting the nucleic acid reporter by HAD. In one example, the method comprises detecting the nucleic acid reporter by fluorescence-based detection. For example, using TaqMan probes, SYBR Green, and molecular beacons. In another example, the method comprises detecting the nucleic acid reporter by microarray techniques.

[0120] In one example, the first capture probe is directly coupled to the first solid matrix. In another example, the second capture probe is directly coupled to the second solid matrix.

[0121] In one example, the first solid matrix and the second solid matrix are the same solid matrix. In another example, the first solid matrix and the second solid matrix are different.

[0122] In one example, the first solid matrix and / or the second solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multi-well plate, an affinity column, a membrane or combinations thereof. In another example, the first solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multi-well plate, an affinity column, a membrane or combinations thereof. In a further example, the second solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multi-well plate, an affinity column, a membrane or combinations thereof.

[0123] In one example, the solid matrix is a magnetic bead or particle. In one example, the solid matrix is a paramagnetic bead or particle. In a further example, the solid matrix is a well of a multiwell plate. In a further example, the solid matrix is an affinity column. In one example, the solid matrix is a membrane. For example, the membrane is a PDVF membrane or a nitrocellulose membrane.

[0124] In one example, the method comprises forming the immunocomplex in a solution prior to capturing the immunocomplex on the first solid matrix.

[0125] In one example, the immunocomplex is formed in a solution and captured on the first solid matrix simultaneously.

[0126] In one example, the first and second binding domains of the target ligand are identical sequences, are different sequences and / or have at least 75% sequence identity. In another example, the first and second binding domains of the target ligand are identical sequences. In another example, the first and second binding domains of the target ligand are different sequences. In a further example, the first and second binding domains of the target ligand have at least 75% sequence identity. For example, about 75%, or about 76%, or about 77%, or about 78%, or about 79%. In one example, the first and second binding domains of the target ligand have at least 80% sequence identity. For example, about 80%, or about 81%, or about 82%, or about 83%, or about 84%. In another example, the first and second binding domains of the target ligand have at least 85% sequence identity. For example, about 85%, or about 86%, or about 87%, or about 88%, or about 89%. In one example, the first and second binding domains of the target ligand have at least 90% sequence identity. For example, about 90%, or about 91%, or about 92%, or about 93%, or about 94%. In another example, the first and second binding domains of the target ligand have at least 95% sequence identity. For example, about 95%, or about 96%, or about 97%, or about 98%, or about 99%.

[0127] In one example, the first receptor, fragment, or epitope thereof and the second receptor, fragment or epitope thereof are identical receptors, fragments or epitopes thereof.

[0128] In one example, the first and / or second receptor, fragment or epitope thereof is selected from the group consisting of a Fc receptor, fragment or epitope thereof; a T cell receptor, fragment or epitope thereof; aB cell receptor, fragment or epitope thereof; an epidermal growth factor (EGF) receptor, fragment or epitope thereof; a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof; an insulin receptor, fragment or epitope thereof; a programmed death 1 (PD-1) receptor, fragment or epitope thereof; and combinations thereof.

[0129] In one example, the first and / or second receptor, fragment, or epitope thereof is a Fc receptor. In one example, the first receptor, fragment, or epitope thereof is a Fc receptor. In another example, the second receptor, fragment, or epitope thereof is a Fc receptor. In a further example, the first and second receptors, fragments or epitopes thereof are Fc receptors.

[0130] In one example, the Fc receptor, fragment, or epitope thereof is a Fc-gamma receptor (FcyR), a Fc-alpha receptor (FcaR), a Fc-epsilon receptor (FcsR) and / or a polymeric immunoglobulin receptor (plgR). In another example, the Fc receptor, fragment or epitope thereof is a FcyR. For example, the FcyR is a FcyRI, a FcyRIIA, a FcyRIIB, a FcyRIIIA, or a FcyRIIIB.

[0131] In one example, the FcsRI is an a-chain of FcsRI or subunit thereof, or a P-chain of FcsRI or subunit thereof.

[0132] In one example, the first receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof.

[0133] In one example, the first receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof.

[0134] In one example, the first receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof.

[0135] In one example, the first receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof.

[0136] In one example, the first and / or second receptor, fragment or epitope thereof is a T cell receptor, fragment or epitope thereof. In another example, the first receptor, fragment or epitope thereof is a T cell receptor, fragment or epitope thereof. In a further example, the second receptor, fragment or epitope thereof is a T cell receptor, fragment or epitope thereof. In one example, the first and / or second receptor, fragment or epitope thereof is a B cell receptor, fragment or epitope thereof. In another example, the first receptor, fragment or epitope thereof is a B cell receptor. In a further example, the second receptor, fragment or epitope thereof is a B cell receptor.

[0137] In one example, the first and / or second receptor, fragment or epitope thereof is an EGF receptor, fragment or epitope thereof. In another example, the first receptor, fragment or epitope thereof is an EGF receptor, fragment or epitope thereof. In a further example, the second receptor, fragment or epitope thereof is an EGF receptor, fragment or epitope thereof.

[0138] In one example, the first and / or second receptor, fragment or epitope thereof is a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof. In another example, the first receptor, fragment or epitope thereof is a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof. In a further example, the second receptor, fragment or epitope thereof is a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof. For example, the TNFR superfamily, fragment or epitope thereof is a TNFR1, a TNFR2, lymphotoxin beta receptor, 0X40, cluster of differentiation (CD) 40 (CD40), Fas receptor, a decoy receptor, CD27, CD30, 4-1BB, a death receptor, receptor activator of nuclear factor K B (RANK), osteoprotegerin, TNF-related weak inducer of apoptosis (TWEAK) receptor, transmembrane activator and CAME interactor (TACI), B-cell activating factor (BAFF) receptor and / or herpesvirus entry mediator.

[0139] In one example, the first and / or second receptor, fragment or epitope thereof is a TNFR1. In one example, the first receptor, fragment or epitope thereof is a TNFR1. In one example, the second receptor, fragment or epitope thereof is a TNFR1.

[0140] In one example, the first and / or second receptor, fragment or epitope thereof is a TNFR2. In one example, the first receptor, fragment or epitope thereof is a TNFR2. In one example, the second receptor, fragment or epitope thereof is a TNFR2.

[0141] In one example, the first and / or second receptor, fragment or epitope thereof is a lymphotoxin beta receptor. In one example, the first receptor, fragment or epitope thereof is a lymphotoxin beta receptor. In one example, the second receptor, fragment or epitope thereof is a lymphotoxin beta receptor.

[0142] In one example, the first and / or second receptor, fragment or epitope thereof is a 0X40. In one example, the first receptor, fragment or epitope thereof is a 0X40. In one example, the second receptor, fragment or epitope thereof is a 0X40.

[0143] In one example, the first and / or second receptor, fragment or epitope thereof is a CD40. In one example, the first receptor, fragment or epitope thereof is a CD40. In one example, the second receptor, fragment or epitope thereof is a CD40.

[0144] In one example, the first and / or second receptor, fragment or epitope thereof is a Fas receptor. In one example, the first receptor, fragment or epitope thereof is a Fas receptor. In one example, the second receptor, fragment or epitope thereof is a Fas receptor.

[0145] In one example, the first and / or second receptor, fragment or epitope thereof is a decoy receptor. In one example, the first receptor, fragment or epitope thereof is a decoy receptor. In one example, the second receptor, fragment or epitope thereof is a decoy receptor. In one example, the first and / or second receptor, fragment or epitope thereof is a CD27. In one example, the first receptor, fragment or epitope thereof is a CD27. In one example, the second receptor, fragment or epitope thereof is a CD27.

[0146] In one example, the first and / or second receptor, fragment or epitope thereof is a CD30. In one example, the first receptor, fragment or epitope thereof is a CD30. In one example, the second receptor, fragment or epitope thereof is a CD30.

[0147] In one example, the first and / or second receptor, fragment or epitope thereof is a 4- IBB. In one example, the first receptor, fragment or epitope thereof is a 4-1BB. In one example, the second receptor, fragment or epitope thereof is a 4- IBB (CD 137).

[0148] In one example, the first and / or second receptor, fragment or epitope thereof is a death receptor. In one example, the first receptor, fragment or epitope thereof is a death receptor. In one example, the second receptor, fragment or epitope thereof is a death receptor.

[0149] In one example, the first and / or second receptor, fragment or epitope thereof is RANK. In one example, the first receptor, fragment or epitope thereof is RANK. In one example, the second receptor, fragment or epitope thereof is RANK.

[0150] In one example, the first and / or second receptor, fragment or epitope thereof is osteoprotegerin. In one example, the first receptor, fragment or epitope thereof is osteoprotegerin. In one example, the second receptor, fragment or epitope thereof is osteoprotegerin.

[0151] In one example, the first and / or second receptor, fragment or epitope thereof is TNF-related weak inducer of apoptosis (TWEAK) receptor. In one example, the first receptor, fragment or epitope thereof is TNF-related weak inducer of apoptosis (TWEAK) receptor. In one example, the second receptor, fragment or epitope thereof is TNF-related weak inducer of apoptosis (TWEAK) receptor.

[0152] In one example, the first and / or second receptor, fragment or epitope thereof is transmembrane activator and CAML interactor (TACI). In one example, the first receptor, fragment or epitope thereof is transmembrane activator and CAML interactor (TACI). In one example, the second receptor, fragment or epitope thereof is transmembrane activator and CAML interactor (TACI).

[0153] In one example, the first and / or second receptor, fragment or epitope thereof is B-cell activating factor (BAFF) receptor. In one example, the first receptor, fragment or epitope thereof is B-cell activating factor (BAFF) receptor. In one example, the second receptor, fragment or epitope thereof is B-cell activating factor (BAFF) receptor.

[0154] In one example, the first and / or second receptor, fragment or epitope thereof is a herpesvirus entry mediator. In one example, the first receptor, fragment or epitope thereof is a herpesvirus entry mediator. In one example, the second receptor, fragment or epitope thereof is a herpesvirus entry mediator.

[0155] In one example, the first and / or second receptor, fragment or epitope thereof is an insulin receptor, fragment or epitope thereof. In one example, the first receptor, fragment or epitope thereof is an insulin receptor, fragment or epitope thereof. In one example, the second receptor, fragment or epitope thereof is an insulin receptor, fragment or epitope thereof. In one example, the first and / or second receptor, fragment or epitope thereof is a programmed death 1 (PD-1) receptor, fragment or epitope thereof. In one example, the first receptor, fragment or epitope thereof is a programmed death 1 (PD-1) receptor, fragment or epitope thereof. In one example, the second receptor, fragment or epitope thereof is a programmed death 1 (PD-1) receptor, fragment or epitope thereof.

[0156] In one example, the target ligand is an immunoglobulin (Ig), a peptide-MHC complex and / or an antigen. In another example, the target ligand is an Ig. In a further example, the target ligand is a peptide-MHC complex. In one example, the target ligand is an antigen.

[0157] In one example, the immunoglobulin is an anti-Fc receptor Ig. For example, the anti-Fc receptor Ig is an anti-FcyR Ig, an anti-FcaR Ig or an anti-FcsR Ig.

[0158] In one example, the anti-Fc receptor Ig is an anti-FcyR Ig. For example, the anti-FcyR Ig is an anti-FcyRI Ig, an anti-FcyRIIA Ig, an anti-FcyRIIB Ig, an anti-FcyRIIIA Ig, or an anti-FcyRIIIB Ig. In one example, the anti-FcyR Ig is an anti-FcyRI Ig. In another example, the anti-FcyR Ig is an anti-FcyRIIA Ig. In a further example, the anti-FcyR Ig is an anti-FcyRIIB Ig. In one example, the anti-FcyR Ig is an anti-FcyRIIIA Ig. In another example, the anti-FcyR Ig is an anti-FcyRIIIB Ig

[0159] In one example, the anti-Fc receptor Ig is an anti-FcaR Ig. For example, the anti-FcaR Ig is an anti-FcaRI Ig.

[0160] In one example, the anti-Fc receptor Ig is an anti-FcaR Ig. For example, the anti-FcaR Ig is an anti-FcaRI Ig or an anti-FcsRII Ig. In one example, the anti-FcaR Ig is an anti-FcaRI Ig. In another example, the the anti-FcaR Ig is an anti-FcsRII Ig.

[0161] In one example, the antigen is selected from the group consisting of an EGF, a programmed death ligand 1 (PD-L1), a PD-L2, a TNF-a, a TNF-C, an OX40L, a CD154, a FasL, a CD70, a Siva, a CD153, a 4-1BB ligand, a TNF-related apoptosis inducing ligand (TRAIL), a RANK ligand (RANKL), TWEAK, a proliferation inducing ligand (APRIL), BAFF, a calcium modulating ligand (CAMLG), LIGHT, TNF-like ligand 1 A (TL1 A) and combinations thereof.

[0162] In one example, the antigen is an EGF. In another example, the antigen is PD-L1. In a further example, the antigen is PD-L2. In one example, the antigen is a TNF-a. In another example, the antigen is a TNF-C. In one example, the antigen is an OX40L. In another example, the antigen is a CD154. In a further example, the antigen is a FasL. In one example, the antigen is a Siva. In another example, the antigen is a CD 153. In a further example, the antigen is a 4- IBB ligand. In one example, the antigen is a TRAIL. In another example, the antigen is a RANKL. In a further example, the antigen is a RANKL. In one example, the antigen is a TWEAK. In one example, the antigen is an APRIL. In another example, the antigen is BAFF. In a further example, the antigen is a CAMLG. In one example, the antigen is LIGHT. In another example, the antigen is TL1 A.

[0163] The present disclosure provides a method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample, the method comprising:

[0164] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0165] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0166] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0167] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0168] The present disclosure provides a method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample, the method comprising:

[0169] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0170] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0171] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0172] The present disclosure provides a method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample, the method comprising:

[0173] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0174] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0175] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0176] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0177] The present disclosure provides a method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample, the method comprising:

[0178] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0179] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0180] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0181] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0182] The present disclosure provides a method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample, the method comprising:

[0183] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0184] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0185] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0186] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0187] The present disclosure further provides a method of screening a sample for an anti-Fc- epsilon receptor I (FcsRI) immunoglobulin (Ig), the method comprising:

[0188] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0189] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0190] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0191] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0192] The present disclosure further provides a method of screening a sample for an anti-Fc- epsilon receptor I (FcsRI) immunoglobulin (Ig), the method comprising:

[0193] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0194] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0195] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0196] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0197] The present disclosure further provides a method of screening a sample for an anti-Fc- epsilon receptor I (FcsRI) immunoglobulin (Ig), the method comprising: (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0198] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0199] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0200] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0201] The present disclosure further provides a method of screening a sample for an anti-Fc- epsilon receptor I (FcsRI) immunoglobulin (Ig), the method comprising:

[0202] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0203] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag; (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0204] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0205] The present disclosure further provides a method of screening a sample for an anti-Fc- epsilon receptor I (FcsRI) immunoglobulin (Ig), the method comprising:

[0206] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0207] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0208] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0209] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0210] The present disclosure also provides a method of identifying a sample suitable for administration to a subject, the method comprising:

[0211] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0212] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0213] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0214] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0215] The present disclosure also provides a method of identifying a sample suitable for administration to a subject, the method comprising:

[0216] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0217] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0218] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0219] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0220] The present disclosure also provides a method of identifying a sample suitable for administration to a subject, the method comprising:

[0221] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0222] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0223] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0224] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0225] The present disclosure also provides a method of identifying a sample suitable for administration to a subject, the method comprising:

[0226] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0227] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag; (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0228] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0229] The present disclosure also provides a method of identifying a sample suitable for administration to a subject, the method comprising:

[0230] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0231] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0232] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0233] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0234] The present disclosure further provides a method of screening a sample to determine suitability of the sample for administration to a subject, the method comprising:

[0235] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0236] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0237] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0238] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0239] The present disclosure further provides a method of screening a sample to determine suitability of the sample for administration to a subject, the method comprising:

[0240] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0241] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0242] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0243] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0244] The present disclosure further provides a method of screening a sample to determine suitability of the sample for administration to a subject, the method comprising:

[0245] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0246] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0247] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0248] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0249] The present disclosure further provides a method of screening a sample to determine suitability of the sample for administration to a subject, the method comprising:

[0250] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI P-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0251] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag; (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0252] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0253] The present disclosure further provides a method of screening a sample to determine suitability of the sample for administration to a subject, the method comprising:

[0254] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a FcsRI P-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0255] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0256] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0257] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0258] In one example of any of the preceding 20 paragraphs, the method further provides performing one or more examples described herein.

[0259] In one example, the sample is serum, plasma, a plasma fraction, a purified or partially purified plasma or plasma fraction thereof, a fermentation broth or a purified or partially purified fermentation broth thereof, a cell culture harvest or a purified or partially purified cell culture harvest thereof or a protein suspension.

[0260] In one example, the plasma or plasma fraction is selected the group consisting of a human blood plasma sample, an IgG intermediate product, a cryo-rich plasma, a cryo-poor plasma, a Supernatant I (SN I), a Cohn Fraction II (Fr II), a Cohn Fraction II+III (Fr II+III), a Cohn Fraction (I+)II+III ((FrI+)II+III), a Kistler / Nitschmann Precipitate A (KN A), a Kistler / Nitschmann Precipitate B (KN B), a Kistler / Nitschmann Precipitate of Supernatant B (KN B+l), an IgG depleted intermediate product, a purified IgG product, a purified formulated IgG product and combinations thereof. In one example, the plasma or plasma fraction is a human blood plasma sample. In another example, the plasma fraction is an IgG intermediate product. In one example, the plasma fraction is cryo-rich plasma. In another example, the plasma fraction is cryo-poor plasma. In a further example, the plasma fraction is Supernatant I (SN I). In one example, the plasma fraction is Cohn Fraction II (Fr II). In another example, the plasma faction is Cohn Fraction II+III (Fr II+III). In a further example, the plasma fraction is Cohn Fraction (I+)II+III ((FrI+)II+III). In one example, the plasma fraction is Kistler / Nitschmann Precipitate A (KN A). In another example, the plasma fraction is Kistler / Nitschmann Precipitate B (KN B). In a further example, the plasma fraction is Kistler / Nitschmann Precipitate of Supernatant B (KN B+l). In one example, the plasma fraction is an IgG depleted intermediate product. In one example, the plasma or plasma fraction is a purified IgG product. In another example, the plasma or plasma fraction is a purified formulated IgG product.

[0261] In one example, the plasma fraction is a suspended paste. For example, the suspended paste is selected from a group consisting of Cohn Fraction II (Fr II), Cohn Fraction II+III (Fr II+III), Cohn Fraction (I+)II+III ((FrI+)II+III), Kistler / Nitschmann Precipitate A (KN A), Kistler / Nitschmann Precipitate B (KN B), Kistler / Nitschmann Precipitate of Supernatant B (KN B+l), and combinations thereof. In one example, the suspended paste is a Cohn Fraction II (Fr II) paste. In one example, the suspended paste is a Cohn Fraction II+III (Fr II+III) paste. In another example, the suspended paste is a Cohn Fraction (I+)II+III ((FrI+)II+III) paste. In another example, the suspended paste is a Kistler / Nitschmann Precipitate A (KN A) paste. In another example, the suspended paste is a Kistler / Nitschmann Precipitate B (KN B) paste. In a further example, the suspended paste is a Kistler / Nitschmann Precipitate of Supernatant B (KN B+l) paste.

[0262] In one example, the plasma fraction is selected from the group consisting of a mammalian plasma fraction, a human plasma fraction, an equine plasma fraction, and a bovine plasma fraction. In one example, the plasma fraction is a mammalian plasma fraction. In one example, the plasma fraction is a human plasma fraction. In one example, the plasma fraction is an equine plasma fraction. In one example the plasma fraction is a bovine plasma fraction. In one example the plasma fraction is a bovine plasma fraction comprising human polyclonal antibodies.

[0263] In one example, the plasma or plasma fraction has been purified or partially purified.

[0264] In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to one or more steps selected from the group consisting of clarification, ethanol precipitation, octanoic acid fractionation, ammonium sulphate precipitation, affinity chromatography, ion exchange chromatography, viral inactivation, viral filtration, isoagglutinin affinity chromatography, ultrafiltration / diafiltration, bulk formulation, final formulation and combinations thereof.

[0265] In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to clarification. In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to ethanol precipitation. In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to octanoic acid fractionation. In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to ammonium sulphate precipitation.

[0266] In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to affinity chromatography. For example, the purified or partially purified plasma or plasma fraction thereof has been subjected to an affinity chromatography resin comprising a ligand capable of specifically binding to a CH3 domain of human IgG. In one example, the purified or partially purified plasma or plasma fraction thereof been subjected to continuous affinity chromatography, the method comprising binding the IgG to an affinity chromatography resin comprising a ligand capable of specifically binding to a CH3 domain of human IgG and collecting the IgG. In one example, the ligand comprises a camelid-derived single domain [VHH] antibody fragment. In one example, the resin comprises a matrix selected from the group consisting of a cross-linked poly(styrene-divinylbenzene) matrix and an agarose-based matrix. In one example, the ligand comprises a VHH antibody fragment conjugated to a cross-linked poly(styrene- divinylbenzene) matrix.

[0267] In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to isoagglutinin affinity chromatography. In one example, the purified or partially purified plasma or plasma fraction thereof has been subjected to ultrafiltration / diafiltration.

[0268] In one example, the method further comprises one or more steps selected from a group consisting of ethanol precipitation, octanoic acid fractionation, affinity chromatography, viral filtration, ultrafiltration / diafiltration and combinations thereof.

[0269] In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 500 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 450 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 400 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 350 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 300 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 350 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 200 pg / mL. In one example, the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 150 pg / mL.

[0270] In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 500 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 450 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 400 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 350 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 300 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 350 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 200 pg / mL. In one example, the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 150 pg / mL.

[0271] In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 150 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 200 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 250 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 300 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 350 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti- FcsRI Ig is present in the sample at a concentration of greater than 400 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 450 pg / mL. In one example, the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 500 pg / mL.

[0272] The present disclosure provides a method of identifying a sample suitable for use in producing an immunoglobulin (Ig) preparation for administration to a subject, the method comprising:

[0273] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0274] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0275] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0276] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 500 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject.

[0277] In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 450 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 400 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 350 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 300 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 250 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 200 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 150 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject.

[0278] The present disclosure also provides a method of screening a subject to determine suitability for plasma donation for production of an immunoglobulin (Ig) preparation, the method comprising: (i) contacting a sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0279] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0280] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0281] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 500 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0282] The present disclosure further provides a method of identifying a subject suitable for plasma donation for production of an immunoglobulin (Ig) preparation, the method comprising:

[0283] (i) contacting a sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex; (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0284] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0285] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 500 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0286] In one example, the anti-FcsRI Ig in the sample activates a FcsRI mediated signaling pathway if administered to a subject.

[0287] In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 450 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 400 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 350 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 300 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 250 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 200 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. In one example, the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 150 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0288] In one example, anti-FcsRI Ig at a concentration of greater than 500 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 450 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 400 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 350 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 300 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 250 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 200 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, anti-FcsRI Ig at a concentration of greater than 150 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject. In one example, activation of the FcsRI mediated signaling pathway induces basophil activation. For example, basophil activation as measured using the Basophil Activation Assay (BAT).

[0289] In one example, activation of the FcsRI mediated signaling pathway induces mast cell activation.

[0290] In one example, activation of the FcsRI mediated signaling pathway induces basophil degranulation.

[0291] In one example, activation of the FcsRI mediated signaling pathway induces mast cell degranulation.

[0292] In one example, the method of the disclosure is a surrogate basophil activation assay. For example, the method of the disclosure is a surrogate assay for the in vitro basophil activation assay (BAT). In one example, the method of the disclosure is indicative of the ability of the anti-FcsRI Ig in the sample to induce basophil activation and / or degranulation if administered to the subject. In one example, the method of the disclosure is indicative of the ability of the anti-FcsRI Ig in the sample to induce basophil activation and / or degranulation as measured in an in vitro BAT.

[0293] In one example, the method of the disclosure identifies samples comprising a level of anti- FcsRI Ig sufficient to induce basophil activation and / or degranulation if administered to the subject. In one example, the method of the disclosure identifies samples comprising a level of anti- FcsRI Ig sufficient to induce basophil activation and / or degranulation as measured in an in vitro BAT.

[0294] In one example, the method of the disclosure is a surrogate mast cell activation assay. For example, the method of the disclosure is a surrogate assay for the in vitro mast cell activation and degranulation assay. In one example, the method of the disclosure is indicative of the ability of the anti-FcsRI Ig in the sample to induce mast cell activation and / or degranulation if administered to the subject. In one example, the method of the disclosure is indicative of the ability of the anti- FcsRI Ig in the sample to induce mast cell activation and / or degranulation as measured in an in vitro mast cell and degranulation assay.

[0295] In one example, the method of the disclosure identifies samples comprising a level of anti- FcsRI Ig sufficient to induce mast cell activation and / or degranulation if administered to the subject. In one example, the method of the disclosure identifies samples comprising a level of anti- FcsRI Ig sufficient to induce mast cell activation and / or degranulation as measured in an in vitro mast cell and degranulation assay.

[0296] In one example, the anti-FcsRI Ig is an anti-FcsRI IgG, an anti-FcsRI IgE, an anti-FcsRI IgM and / or an anti-FcsRI IgA. In another example, the anti-FcsRI Ig is an anti-FcsRI IgG. In a further example, the anti-FcsRI Ig is an anti-FcsRI IgE. In one example, the anti-FcsRI Ig is an anti-FcsRI IgM. In another example, the anti-FcsRI Ig is an anti-FcsRI IgA.

[0297] In one example, the anti-FcsRI IgG is an IgGl, an IgG2, an IgG3 or an IgG4. In another example, the anti-FcsRI IgG is an IgGl. In a further example, the anti-FcsRI IgG is an IgG2. In one example, the anti-FcsRI IgG is an IgG3. In a further example, the anti-FcsRI IgG is an IgG4. In one example, the method further comprises diluting the sample in an assay solution prior to contacting with the first binding moiety and the second binding moiety.

[0298] In one example, the sample is diluted in the assay solution at a ratio of between 1 : 10 to 1 :20. For example, the sample is diluted in the assay solution at a ratio of about 1 : 10, or about 1 : 11, or about 1 : 12, or about 1 : 13, or about 1 : 14, or about 1 :15, or about 1 : 16, or about 1 : 17, or about 1 : 18, or about 1 : 19, or about 1 :20.

[0299] In one example, the sample is in the assay solution at a concentration of 10% (v / v).

[0300] In one example, the first binding moiety and / or the second binding moiety contacts the sample at a concentration of between 4-fold to 0.0625-fold. For example, the first binding moiety and / or the second binding moiety contacts the sample at a concentration of about 4-fold, or about

[0301] 3-fold, or about 2-fold, or about 1-fold. In another example, first binding moiety and / or the second binding moiety contacts the sample at a concentration of about 0.5-fold, or about 0.25-fold, or about 0.125-fold, or about 0.0625-fold. In one example, first binding moiety and / or the second binding moiety contacts the sample at a concentration of about 0.5-fold. In another example, first binding moiety and / or the second binding moiety contacts the sample at a concentration of about 0.25-fold. In a further example, first binding moiety and / or the second binding moiety contacts the sample at a concentration of about 0.125-fold.

[0302] In one example, the first binding moiety contacts the sample at a concentration of between

[0303] 4-fold to 0.0625-fold. For example, the first binding moiety contacts the sample at a concentration of about 4-fold, or about 3 -fold, or about 2-fold, or about 1-fold. In another example, first binding moiety contacts the sample at a concentration of about 0.5-fold, or about 0.25-fold, or about 0.125- fold, or about 0.0625-fold. In one example, first binding moiety contacts the sample at a concentration of about 0.5-fold. In another example, first binding moiety contacts the sample at a concentration of about 0.25-fold. In a further example, first binding moiety contacts the sample at a concentration of about 0.125-fold.

[0304] In one example, the second binding moiety contacts the sample at a concentration of between 4-fold to 0.0625-fold. For example, the second binding moiety contacts the sample at a concentration of about 4-fold, or about 3 -fold, or about 2-fold, or about 1-fold. In another example, second binding moiety contacts the sample at a concentration of about 0.5-fold, or about 0.25-fold, or about 0.125-fold, or about 0.0625-fold. In one example, second binding moiety contacts the sample at a concentration of about 0.5-fold. In another example, second binding moiety contacts the sample at a concentration of about 0.25-fold. In a further example, second binding moiety contacts the sample at a concentration of about 0.125 -fold.

[0305] In one example, the method of the disclosure detects the target ligand in the sample, wherein the target ligand is in the sample at a concentration of less than 1 pg / mL.

[0306] In one example, the method of the disclosure has a limit of detection of about 0.2 pg target ligand / mL sample.

[0307] In one example, the method of the disclosure has a limit of detection of 1454 aM target ligand. In one example, the method of the disclosure has a sensitivity and specificity allowing at least 3 -fold separation of a sample comprising the target ligand from a sample without the target ligand. For example, at least 4-fold, or at least 5-fold separation of a sample comprising the target ligand from a sample without the target ligand. In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a sample comprising the target ligand from a sample without the target ligand. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50-fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90-fold, or at least 100-fold separation of a sample comprising the target ligand from a sample without the target ligand.

[0308] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 3-fold separation of a plasma sample comprising the target ligand from a plasma sample without the target ligand. For example, at least 4-fold, or at least 5-fold separation of a plasma sample comprising the target ligand from a plasma sample without the target ligand. In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a plasma sample comprising the target ligand from a plasma sample without the target ligand. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50- fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90-fold, or at least 100- fold separation of a plasma sample comprising the target ligand from a plasma sample without the target ligand.

[0309] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a cryo-poor plasma sample comprising the target ligand from a cryopoor plasma sample without the target ligand. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50-fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90-fold, or at least 100-fold separation of a cryo-poor plasma sample comprising the target ligand from a cryo-poor plasma sample without the target ligand.

[0310] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a donor plasma sample comprising the target ligand from a donor plasma sample without the target ligand. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50-fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90-fold, or at least 100-fold separation of a donor plasma sample comprising the target ligand from a donor plasma sample without the target ligand.

[0311] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 3-fold separation of a plasma sample comprising anti-FcsRI Ig from a plasma sample without anti-FcsRI Ig. For example, at least 4-fold, or at least 5-fold separation of a plasma sample comprising anti-FcsRI Ig from a plasma sample without anti-FcsRI Ig. In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a plasma sample comprising anti-FcsRI Ig from a plasma sample without anti-FcsRI Ig. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50-fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90-fold, or at least 100-fold separation of a plasma sample comprising anti-FcsRI Ig from a plasma sample without anti-FcsRI Ig. In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a cryo-poor plasma sample comprising anti-FcsRI Ig from a cryo-poor plasma sample without anti-FcsRI Ig. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50-fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90- fold, or at least 100-fold separation of a cryo-poor plasma sample comprising anti-FcsRI Ig from a cryo-poor plasma sample without anti-FcsRI Ig.

[0312] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 10-fold separation of a donor plasma sample comprising anti-FcsRI Ig from a donor plasma sample without anti-FcsRI Ig. For example, at least 20-fold, or at least 30-fold, or at least 40-fold, or at least 50-fold, or at least 60-fold, or at least 70-fold, or at least 80-fold, or at least 90-fold, or at least 100-fold separation of a donor plasma sample comprising anti-FcsRI Ig from a donor plasma sample without anti-FcsRI Ig.

[0313] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 3 -fold separation of a purified formulated product comprising the target ligand from a purified formulated product without the target ligand. In one example, the method of the disclosure has a sensitivity allowing about 3-fold separation of a purified formulated product comprising the target ligand from a purified formulated product without the target ligand. In one example, the method of the disclosure has a sensitivity and specificity allowing about 4-fold separation of a purified formulated product comprising the target ligand from a purified formulated product without the target ligand.

[0314] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 3-fold separation of a purified formulated Ig product comprising the target ligand from a purified formulated Ig product without the target ligand. In one example, the method of the disclosure has a sensitivity and specificity allowing about 3-fold separation of a purified formulated Ig product comprising the target ligand from a purified formulated Ig product without the target ligand. In one example, the method of the disclosure has a sensitivity and specificity allowing about 4-fold separation of a purified formulated Ig product comprising the target ligand from a purified formulated Ig product without the target ligand.

[0315] In one example, the method of the disclosure has a sensitivity and specificity allowing at least 3-fold separation of a purified formulated Ig product comprising anti-FcsRI Ig from a purified formulated Ig product without anti-FcsRI Ig. In one example, the method of the disclosure has a sensitivity and specificity allowing about 3 -fold separation of a purified formulated Ig product comprising anti-FcsRI Ig from a purified formulated Ig product without anti-FcsRI Ig. In one example, the method of the disclosure has a sensitivity and specificity allowing about 4-fold separation of a purified formulated Ig product comprising anti-FcsRI Ig from a purified formulated Ig product without anti-FcsRI Ig.

[0316] It will be apparent to the skilled person from the disclosure herein that reference to a sample or product “without the target ligand” or “without anti-FcsRI Ig” refers to a sample or product without a level of target ligand and / or anti-FcsRI Ig capable of inducing unwanted adverse effects as described herein. In one example of any method described herein, the subject is a mammal, for example a primate such as a human.

[0317] In one example, the method of the disclosure is performed before the sample comprising the anti-FcsRI Ig is subjected to a chromatography step. For example, the methods of detecting, screening and identifying of the disclosure further comprise preparing an anti-FcsRI Ig reduced preparation from the sample if anti-FcsRI Ig is present in the sample, the method further comprising binding the anti-FcsRI Ig to a chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the sample and collecting the anti-FcsRI Ig reduced preparation.

[0318] In one example, the methods of detecting, screening and identifying of the disclosure further comprise preparing an anti-FcsRI Ig reduced preparation from the sample if anti-FcsRI Ig is present in the sample, the method further comprising:

[0319] (i) loading the sample onto a chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the sample; and

[0320] (ii) collecting the anti-FcsRI Ig reduced preparation.

[0321] In one example, the method of detecting of the disclosure is performed to detect if anti- FcsRI Ig is present in the sample before the sample is subjected to a chromatography step. In one example, the method of screening of the disclosure is performed to screen if anti-FcsRI Ig is present in the sample before the sample is subjected to a chromatography step. In one example, the method of identifying of the disclosure is performed to identify if anti-FcsRI Ig is present in the sample before the sample is subjected to a chromatography step.

[0322] The present disclosure also provides a method of preparing an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation from a plasma sample or a fraction thereof, the method comprising binding the anti-FcsRI Ig to a chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the plasma sample or fraction thereof and collecting the anti-FcsRI Ig reduced preparation.

[0323] The present disclosure also provides a method of preparing an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation from a plasma sample or a fraction thereof, the method comprising:

[0324] (i) loading the plasma sample or fraction thereof onto an chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the plasma sample or fraction thereof; and

[0325] (ii) collecting the anti-FcsRI Ig reduced preparation.

[0326] In one example, the chromatography resin is an affinity chromatography resin. For example, a continuous affinity chromatography resin.

[0327] The present disclosure provides a method of preparing an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation from a plasma sample or a fraction thereof, the method comprising:

[0328] (i) loading the plasma sample or fraction thereof onto an affinity chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the plasma sample or fraction thereof; and (ii) collecting an anti-FcsRI Ig reduced preparation.

[0329] In one example, the ligand is immobilized to a matrix of the chromatography resin.

[0330] In one example, the method of detecting of the disclosure is performed to detect if anti- FcsRI Ig is present in the sample before the sample is loaded onto the chromatography resin. In one example, the method of screening of the disclosure is performed to screen if anti-FcsRI Ig is present in the sample before the sample is loaded onto the chromatography resin. In one example, the method of identifying of the disclosure is performed to identify if anti-FcsRI Ig is present in the sample before the sample is loaded onto the chromatography resin.

[0331] In one example, the ligand comprises FcsRI or fragment or epitope thereof. For example, the fragment of FcsRI comprises an a-chain of FcsRI. In one example, the FcsRI fragment is a functional fragment. For example, the FcsRI functional fragment is a fragment of FcsRI that, when bound to the anti-FcsRI Ig, activates a FcsRI mediated signalling pathway.

[0332] In one example, the anti-FcsRI Ig reduced preparation induces reduced activation of a FcsRI mediated signalling pathway relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation does not induce activation of a FcsRI mediated signalling pathway.

[0333] In one example, the anti-FcsRI Ig reduced preparation comprises less than 500 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 450 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 400 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 350 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 300 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 250 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 200 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 150 pg / mL of anti-FcsRI Ig. In one example, the anti- FcsRI Ig reduced preparation comprises less than 100 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 50 pg / mL of anti-FcsRI Ig.

[0334] In one example, the method comprises determining the level of anti-FcsRI Ig in the anti- FcsRI Ig reduced preparation. For example, the method comprises determining if a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation. In one example, if the anti-FcsRI IgG is not present in a detectable amount in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 500 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 450 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 400 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 350 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 300 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 250 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 200 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject. In one example, if the anti-FcsRI IgG is present at less than 150 pg / mL in the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced preparation is suitable for administration to the subject.

[0335] In one example, the anti-FcsRI Ig reduced preparation induces reduced basophil and mast cell activation and / or degranulation relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation induces reduced basophil activation relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation induces reduced mast cell activation relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation does not induce basophil and / or mast cell activation and / or degranulation.

[0336] In one example, the basophil and / or mast cell activation and / or degranulation following exposure to the anti-FcsRI Ig reduced preparation is determined by a method described herein. For example, the method comprises determining the proportion of basophil and / or mast cells expressing one or more markers selected from the group consisting of CD63, CD203c, CD107a and combinations thereof. In another example, the basophil activation following exposure to the anti-FcsRI Ig reduced preparation is determined by determining the proportion of basophil expressing one or more markers selected from the group consisting of CD63, CD203c and combinations thereof. In a further example, the mast cell activation following exposure to the anti- FcsRI Ig reduced preparation is determined by determining the proportion of mast cells expressing CD 107a.

[0337] In one example, the method comprises determining the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced preparation. For example, the method comprises determining whether a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation by determining the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced preparation. In one example, the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced preparation is less than 5%. For example, the proportion of basophils expressing CD63 is relative to the total proportion of basophils being exposed to the anti-FcsRI Ig reduced preparation. In one example, a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation if at least 5% of a population of basophils express CD63 following exposure to the anti-FcsRI Ig reduced preparation. In one example, no detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation if less than 5% of a population of basophils express CD63 following exposure to the anti-FcsRI Ig reduced preparation.

[0338] In one example, the method comprises determining the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced preparation. For example, the method comprises determining whether a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation by determining the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced preparation. In one example, the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced preparation is less than 5%. For example, the proportion of basophils expressing CD203c is relative to the total proportion of basophils being exposed to the anti-FcsRI Ig reduced preparation. In one example, a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation if at least 5% of a population of basophils express CD203c following exposure to the anti-FcsRI Ig reduced preparation. In one example, no detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation if less than 5% of a population of basophils express CD203c following exposure to the anti-FcsRI Ig reduced preparation.

[0339] In one example, the method comprises determining the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced preparation. For example, the method comprises determining whether a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation by determining the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced preparation. In one example, the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced preparation is less than 5%. For example, the proportion of mast cells expressing CD107a is relative to the total proportion of mast cells being exposed to the anti-FcsRI Ig reduced preparation. In one example, a detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation if at least 5% of a population of mast cells express CD 107a following exposure to the anti-FcsRI Ig reduced preparation. In one example, no detectable amount of anti-FcsRI Ig is present in the anti-FcsRI Ig reduced preparation if less than 5% of a population of mast cells express CD 107a following exposure to the anti-FcsRI Ig reduced preparation.

[0340] In one example, the anti-FcsRI Ig reduced preparation comprises less than 40 pg / mL of an anti-FcsRI IgG and less than 0.1 kU / L of an anti-FcsRI IgE.

[0341] In one example, the method comprises determining the level of anti-FcsRI IgG in the anti- FcsRI Ig reduced preparation. For example, the method comprises determining if a detectable amount of anti-FcsRI IgG is present in the anti-FcsRI Ig reduced preparation. In one example, the detectable amount of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is at least 40 pg / mL. In another example, the anti-FcsRI Ig reduced preparation comprises less than 40 pg / mL of an anti-FcsRI IgG. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 35 pg / mL. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 30 pg / mL. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 35 pg / mL. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 20 pg / mL. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 15 pg / mL. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 10 pg / mL. In one example, the level of anti-FcsRI IgG in the anti-FcsRI Ig reduced preparation is less than 5 pg / mL. In one example, if the anti-FcsRI IgG is present in the anti-FcsRI Ig reduced preparation at a concentration of less than 40 pg / mL (i.e., not in a detectable amount), the anti-FcsRI Ig reduced preparation is suitable for administration to the subject.

[0342] In one example, the method comprises determining the level of anti-FcsRI IgE in the anti- FcsRI Ig reduced preparation. For example, the method comprises determining if a detectable amount of anti-FcsRI IgE is present in the anti-FcsRI Ig reduced preparation. In one example, the detectable amount of anti-FcsRI IgE in the anti-FcsRI Ig reduced preparation is at least 0.1 kU7L. In one example, the anti-FcsRI Ig reduced preparation comprises less than 0.1 kU / L of an anti- FcsRI IgE. In one example, the level of anti-FcsRI IgE in the anti-FcsRI Ig reduced preparation is less than 0.01 kU / L. In one example, the level of anti-FcsRI IgE in the anti-FcsRI Ig reduced preparation is less than 0.05 kU / L. In one example, the level of anti-FcsRI IgE in the anti-FcsRI Ig reduced preparation is less than 0.001 kU / L. In one example, the level of anti-FcsRI IgE in the anti-FcsRI Ig reduced preparation is less than 0.005 kU / L. In one example, if the anti-FcsRI IgE is present in the anti-FcsRI Ig reduced preparation at a concentration of less than 0.1 kU / L (i.e., not in a detectable amount), the anti-FcsRI Ig reduced preparation is suitable for administration to the subject.

[0343] In one example, the anti-FcsRI Ig reduced preparation is an anti-FcsRI Ig reduced IgG preparation.

[0344] In one example, the anti-FcsRI Ig reduced preparation induces reduced basophil and mast cell activation and / or degranulation relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation induces reduced basophil activation relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation induces reduced mast cell activation relative to a preparation wherein the level of the anti-FcsRI Ig is not reduced. In one example, the anti-FcsRI Ig reduced preparation does not induce basophil and / or mast cell activation and / or degranulation.

[0345] In one example, the affinity chromatography resin further comprises a blood group A antigen and a blood group B antigen immobilized to the matrix of the affinity chromatography resin. In another example, the affinity chromatography resin further comprises a blood group A antigen. In a further example, the affinity chromatography resin further comprises a blood group B antigen.

[0346] In one example, the affinity chromatography resin is a continuous affinity chromatography resin. For example, the continuous affinity chromatography resin comprises a ligand which binds to anti-FcsRI Ig.

[0347] In one example, the affinity chromatography resin is a continuous affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig, a blood group A antigen and a blood group B antigen.

[0348] In one example, the affinity chromatography resin is a continuous affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig and a blood group A antigen immobilized to a matrix of the affinity chromatography resin. In one example, the affinity chromatography resin is a continuous affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig and a blood group B antigen immobilized to a matrix of the affinity chromatography resin.

[0349] The present disclosure provides an affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig and optionally, a blood group A antigen and / or a blood group B antigen. For example, the disclosure provides a continuous affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig and optionally, a blood group A antigen and / or a blood group B antigen.

[0350] In one example, the affinity chromatography is continuous affinity chromatography comprising:

[0351] (i) a first affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig immobilized to a matrix of the first affinity chromatography resin; and

[0352] (ii) a second affinity chromatography resin comprising a blood group A antigen and a blood group B antigen immobilized to a matrix of the second affinity chromatography resin. In one example, the affinity chromatography is continuous affinity chromatography comprising:

[0353] (i) a first affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig immobilized to a matrix of the first affinity chromatography resin; and

[0354] (ii) a second affinity chromatography resin comprising a blood group A antigen immobilized to a matrix of the second affinity chromatography resin.

[0355] In one example, the affinity chromatography is continuous affinity chromatography comprising:

[0356] (i) a first affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig immobilized to a matrix of the first affinity chromatography resin; and

[0357] (ii) a second affinity chromatography resin comprising a blood group B antigen immobilized to a matrix of the second affinity chromatography resin.

[0358] In one example, the affinity chromatography is continuous affinity chromatography comprising:

[0359] (i) a first affinity chromatography resin comprising a blood group A antigen and a blood group B antigen immobilized to a matrix of the first affinity chromatography resin; and

[0360] (ii) a second affinity chromatography resin comprising a ligand which binds to anti- FcsRI Ig immobilized to a matrix of the second affinity chromatography resin.

[0361] In one example, the affinity chromatography is continuous affinity chromatography comprising:

[0362] (i) a first affinity chromatography resin comprising a blood group A antigen immobilized to a matrix of the first affinity chromatography resin; and

[0363] (ii) a second affinity chromatography resin comprising a ligand which binds to anti- FcsRI Ig immobilized to a matrix of the second affinity chromatography resin.

[0364] In one example, the affinity chromatography is continuous affinity chromatography comprising: (i) a first affinity chromatography resin comprising a blood group B antigen immobilized to a matrix of the first affinity chromatography resin; and

[0365] (ii) a second affinity chromatography resin comprising a ligand which binds to anti- FcsRI Ig immobilized to a matrix of the second affinity chromatography resin.

[0366] In one example, the ligand comprises FcsRI or fragment or epitope thereof. For example, the fragment of FcsRI comprises an a-chain of FcsRI. In one example, the FcsRI fragment is a functional fragment. For example, the FcsRI functional fragment is a fragment of FcsRI that when bound to the anti-FcsRI Ig activates a FcsRI mediated signalling pathway.

[0367] In one example, the method further comprises one or more steps selected from a group consisting of ethanol precipitation, octanoid acid fractionation, ion exchange chromatography, viral inactivation, viral filtration, ultrafiltration / diafiltration and combinations thereof. The findings further provide the basis for a pharmaceutical composition comprising the anti-FcsRI Ig reduced preparation or the anti-FcsRI Ig reduced IgG plasma preparation, comprising polyclonal IgG, as well as the use of the pharmaceutical composition for treating, preventing and / or delaying progression of a condition (e.g., primary immunodeficiency disease, chronic inflammatory demyelinating polyneuropathy, and chronic immune thrombocytopenic purpura) in a subject.

[0368] The present disclosure provides a pharmaceutical composition comprising an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced IgG preparation produced by a method described herein.

[0369] In one example, the polyclonal IgG is selected from the group consisting of Hizentra® (CSL Behring), Privigen® (CSL Behring), Flebogamma® (Grifols), Gamunex®-C (Grifols), Gammagard® (Takeda), and Octagam® (Octapharma). Gamunex®-C (Grifols), Xembify® (Grifols), Cutaquig® (Octapharma) and Cuvitru® (Takeda).

[0370] In one example, the anti-FcsRI Ig reduced preparation comprises an undetectable amount of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 500 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 450 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 400 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 350 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 300 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 250 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 200 pg / mL of anti-FcsRI Ig. In one example, the anti- FcsRI Ig reduced preparation comprises less than 150 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 100 pg / mL of anti-FcsRI Ig. In one example, the anti-FcsRI Ig reduced preparation comprises less than 50 pg / mL of anti-FcsRI Ig.

[0371] For example, the anti-FcsRI Ig reduced preparation comprises less than 40 pg / mL of anti-FcsRI IgG and / or less than 0.1 kU / L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced preparation comprises less than 40 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 35 pg / mL of anti-FcsRI IgG. In one example, the anti- FcsRI Ig reduced preparation comprises less than 30 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 25 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 20 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 15 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 10 pg / mL of anti- FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 5 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced preparation comprises less than 0.1 kU7L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced preparation comprises less than 0.01 kU / L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced preparation comprises less than 0.005 kU / L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced preparation comprises less than 0.001 kU / L of anti-FcsRI IgE.

[0372] In one example, the anti-FcsRI Ig reduced IgG preparation comprises an undetectable amount of anti-FcsRI Ig. For example, the anti-FcsRI Ig reduced IgG preparation comprises less than 40 pg / mL of anti-FcsRI IgG and / or less than 0.1 kU / L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 40 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 35 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 30 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 25 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 20 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 15 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 10 pg / mL of anti-FcsRI IgG. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 5 pg / mL of anti-FcsRI IgG. In one example, the anti- FcsRI Ig reduced IgG preparation comprises less than 0.1 kU7L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 0.01 kU / L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 0.005 kU / L of anti-FcsRI IgE. In one example, the anti-FcsRI Ig reduced IgG preparation comprises less than 0.001 kU / L of anti-FcsRI IgE.

[0373] In one example, the anti-FcsRI Ig reduced preparation does not induce activation of the FcsRI mediated signalling pathway. For example, the anti-FcsRI Ig reduced preparation does not induce basophil and / or mast cell activation and / or degranulation. In one example, the anti-FcsRI Ig reduced preparation does not induce a detectable amount of basophil and / or mast cell activation and / or degranulation.

[0374] It will be apparent from the disclosure herein that basophil and / or mast cell activation and / or degranulation is determined by determining the proportion of basophils expressing CD63 and / or CD203c and / or mast cells expressing CD107a.In one example, the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5%, the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% and / or the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils and / or mast cells exposed to the anti-FcsRI Ig reduced IgG preparation. In another example, the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti- FcsRI Ig reduced IgG preparation. In a further example, the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcsRI Ig reduced IgG preparation. In one example, the proportion of mast cells expressing CD107a following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of mast cells exposed to the anti- FcsRI Ig reduced IgG preparation. In one example, the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcsRI Ig reduced IgG preparation and / or the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcsRI Ig reduced IgG preparation. In one example, the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcsRI Ig reduced IgG preparation and / or the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of mast cells exposed to the anti-FcsRI Ig reduced IgG preparation. In one example, the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti- FcsRI Ig reduced IgG preparation and / or the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of mast cells exposed to the anti-FcsRI Ig reduced IgG preparation. In one example, the proportion of basophils expressing CD63 following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcsRI Ig reduced IgG preparation, the proportion of basophils expressing CD203c following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcsRI Ig reduced IgG preparation and / or the proportion of mast cells expressing CD 107a following exposure to the anti-FcsRI Ig reduced IgG preparation is less than 5% of the total population of mast cells exposed to the anti-FcsRI Ig reduced IgG preparation.

[0375] The present disclosure also provides a pooled IgG preparation, wherein the pooled IgG preparation comprises a detectable amount of an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig). It will be apparent to the skilled person from the disclosure herein that the pooled IgG preparation with detectable amount of an anti-FcsRI Ig has not been subjected to any method of the present disclosure. Accordingly, the present disclosure also provides a method of detecting anti-FcsRI Ig in a pooled IgG preparation.

[0376] The present disclosure also provides a method of treating, preventing and / or delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject, the method comprising administering the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein. For example, the disclosure provides a method of treating a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject, the method comprising administering the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein. The disclosure also provides a method of preventing a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject, the method comprising administering the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein. The disclosure further provides a method of delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject, the method comprising administering the anti-FcsRI Ig reduced preparation, the anti- FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein.

[0377] The present disclosure also provides the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for use in treating, preventing and / or delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The disclosure also provides the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for use in treating of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The disclosure also provides the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for use in preventing of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The disclosure also provides the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for use in delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject.

[0378] The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti- FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for treating, preventing and / or delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for treating a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for preventing a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein for delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti- FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein in the manufacture of a medicament for the treatment, prevention and / or delayed progression of an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein in the manufacture of a medicament for the treatment of an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein in the manufacture of a medicament for the prevention of an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure also provides use of the anti-FcsRI Ig reduced preparation, the anti-FcsRI Ig reduced IgG preparation or the pharmaceutical composition described herein in the manufacture of a medicament for delaying progression of an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject.

[0379] The present disclosure also provides a method of preparing an immunoglobulin (Ig) preparation from a plasma sample or a fraction thereof, the Ig preparation having reduced adverse reactions in a subject, the method comprising detecting the presence of anti-FcsRI Ig in the plasma sample or fraction thereof according to any method described herein and producing an Ig preparation having reduced adverse reactions.

[0380] In one example, the condition is selected from the group consisting of primary immunodeficiency disease, chronic inflammatory demyelinating polyneuropathy and chronic immune thrombocytopenic purpura. In one example, the condition is primary immunodeficiency disease. In another example, the condition is chronic inflammatory demyelinating polyneuropathy. In a further example, the condition is chronic immune thrombocytopenic purpura.

[0381] The present disclosure also provides a method of preparing an immunoglobulin (Ig) preparation from a plasma sample or a fraction thereof, the Ig preparation having reduced adverse reactions in a subject, the method comprising determining the proportion of basophil and / or mast cell activation and / or degranulation following exposure to the plasma sample or fraction thereof;

[0382] (i) including the plasma sample or fraction thereof into the Ig preparation if the proportion of basophils expressing CD63 following exposure to the plasma sample or fraction thereof is less than 1%, less than 2%, less than 3%, less than 4% or less than 5% of the total population of basophils exposed to the plasma sample or fraction thereof;

[0383] (ii) excluding the plasma sample or fraction thereof into the Ig preparation if the proportion of basophils expressing CD63 following exposure to the plasma sample or fraction thereof is more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 11%, more than 12%, more than 13%, more than 14% or more than 15% of the total population of basophils exposed to the plasma sample or fraction thereof; (iii) including the plasma sample or fraction thereof into the Ig preparation if the proportion of basophils expressing CD203c following exposure to the plasma sample or fraction thereof is less than 1%, less than 2%, less than 3%, less than 4% or less than 5% of the total population of basophils exposed to the plasma sample or fraction thereof;

[0384] (iv) excluding the plasma sample or fraction thereof into the Ig preparation if the proportion of basophils expressing CD203c following exposure to the plasma sample or fraction thereof is more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 11%, more than 12%, more than 13%, more than 14% or more than 15% of the total population of basophils exposed to the plasma sample or fraction thereof;

[0385] (v) including the plasma sample or fraction thereof into the Ig preparation if the proportion of mast cells expressing CD 107a following exposure to the plasma sample or fraction thereof is less than 1%, less than 2%, less than 3%, less than 4% or less than 5% of the total population of mast cells exposed to the plasma sample or fraction thereof; and / or

[0386] (vi) excluding the plasma sample or fraction thereof into the Ig preparation if the proportion of mast cells expressing CD 107a following exposure to the plasma sample or fraction thereof is more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 11%, more than 12%, more than 13%, more than 14% or more than 15% of the total population of mast cells exposed to the plasma sample or fraction thereof.

[0387] The present disclosure also provides methods for identifying a plasma donation suitable for the production of an Ig preparation.

[0388] BRIEF DESCRIPTION OF THE DRAWINGS

[0389] Figure 1 is a graphical representation showing (A) autoantibody capture-detection pair- wise testing and (B) sample separation of normal and adverse event (AE) plasma in cryo-poor plasma (CPP) or final product.

[0390] Figure 2 is a graphical representation showing standard curve of anti-FcsRIa NULISA assay.

[0391] Figure 3 is a graphical representation showing various conjugate concentration of (A) top performing Cl / Dl capture-detection pair (B) sample separation of normal and AE plasma in CPP or final product.

[0392] Figure 4 is a graphical representation showing plasma samples from different normal donors in comparison with positive control and Donor Z plasma sample using the custom anti- FcsRIa assay.

[0393] Figure 5 is a graphical representation showing anti-FcsRIa NULISA assay result for IgProlO final product with and without known AE signal.

[0394] Figure 6 is a graphical representation showing anti-FcsRIa antibody concentration in IgProlO lots without known AE signal and lots manufactured for Canada. Figure 7 is a graphical representation showing comparison of normal donor plasma and Donor Z plasma in different in vitro assays; The classical Histamine release and Basophil activation test (BAT) allows distinction between normal plasma and AE-causing plasma. The anti- human-FcsRIa ELISA did not distinguish between normal and AE-causing plasma. The lowest panel represents NULISA data, demonstrating clearly -10-100 fold separation between normal and Donor Z plasma.

[0395] Figure 8 is a graphical representation showing NULISA testing of unique 100 individual plasma donations for oligomerizing anti-FcsRIa antibodies.

[0396] Figure 9 is a graphical representation showing NULISA testing of plasma minipools.

[0397] Figure 10 is a graphical representation showing anti-FcsRIa antibody levels in (A) CPP pools and (B) dilution-dependent signal reduction.

[0398] Figure 11 is a graphical representation showing anti-FcsRIa antibody levels in IgProlO.

[0399] Figure 12 is a graphical representation showing depletion of anti-FcsRIa autoantibodies from (A) Donor Z containing plasma and (B) Donor Z containing IgProlO semi-finished product using FcsRIa-resin.

[0400] DETAILED DESCRIPTION

[0401] General

[0402] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or groups of compositions of matter.

[0403] Those skilled in the art will appreciate that the present disclosure is susceptible to variations and modifications other than those specifically described. It is to be understood that the disclosure includes all such variations and modifications. The disclosure also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features.

[0404] The present disclosure is not to be limited in scope by the specific examples described herein, which are intended for the purpose of exemplification only. Functionally equivalent products, compositions and methods are clearly within the scope of the present disclosure.

[0405] Any example of the present disclosure herein shall be taken to apply mutatis mutandis to any other example of the disclosure unless specifically stated otherwise.

[0406] Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (for example, in cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0407] The term “and / or”, e.g., “X and / or Y” shall be understood to mean either “X and Y” or “X or Y” and shall be taken to provide explicit support for both meanings or for either meaning.

[0408] The term “about” in relation to a numerical value x is optional and means, for example, any number within 1%, 5% or 10% of the referenced number. In one example, the term “about” in reference to temperature shall be understood to refer to ± 0.2°C. The term “about” also encompasses the exact number recited.

[0409] Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0410] As used herein the term “derived from” shall be taken to indicate that a specified integer may be obtained from a particular source albeit not necessarily directly from that source.

[0411] Furthermore, as used herein the singular forms of “a”, “and” and “the” include plural references unless the context clearly dictates otherwise.

[0412] Unless otherwise indicated, the terms “first,” “second,” etc. are used herein merely as labels, and are not intended to impose ordinal, positional, or hierarchical requirements on the items to which these terms refer. Moreover, reference to a “second” item does not require or preclude the existence of lower-numbered item (e.g., a “first” item) and / or a higher-numbered item (e.g., a “third” item).

[0413] All publications cited herein are hereby incorporated by reference in their entirety. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

[0414] Any discussion of documents, acts, materials, devices, articles or the like that has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.

[0415] Selected Definitions

[0416] As used herein, the term “detect” or “detecting” refers to determining, measuring or assessing the presence, absence, or amount of the target ligand in the sample.

[0417] As used herein, the term “contact” or “contacting” is used to refer to a direct or indirect interaction or association between a target ligand and a first and second binding moiety described herein (e.g., a receptor, fragment or epitope thereof). For example, a target ligand (such as an anti- Fc receptor Ig) either directly or indirectly binds to a first and second binding moiety (e.g., a first and second FcsRI or fragment or epitope thereof) in a sample (e.g., a plasma sample or fraction thereof).

[0418] As used herein “mast cells” shall be understood to refer to granulated tissue-resident cells known for host cell response, allergic response, and vascular homeostasis.

[0419] As used herein “basophils” shall be understood to refer to the largest type of granulocyte representing about 0.5% to 1% of circulating white blood cells. Their activation and degranulation are involved in inflammatory reactions during immune response, as well as in the formation of acute and chronic allergic diseases.

[0420] The term “protein” shall be taken to include a single polypeptide chain, i.e., a series of contiguous amino acids linked by peptide bonds or a series of polypeptide chains covalently or non-covalently linked to one another (i.e., a polypeptide complex). For example, the series of polypeptide chains can be covalently linked using a suitable chemical or a disulfide bond. Examples of non-covalent bonds include hydrogen bonds, ionic bonds, Van der Waals forces, and hydrophobic interactions.

[0421] The term “polypeptide” or “polypeptide chain” will be understood from the foregoing paragraph to mean a series of contiguous amino acids linked by peptide bonds.

[0422] As used herein, the term “nucleotide” or “nucleic acid” will be understood to mean a series of contiguous nucleotides (or bases) covalently linked to a phosphodiester backbone.

[0423] As used herein, the term “epitope” (syri. “antigenic determinant”) shall be understood to mean a region of the receptor or fragment thereof to which a binding domain of a target ligand binds. This term is not necessarily limited to the specific residues or structure to which the receptor or fragment thereof makes contact. For example, this term includes the region spanning amino acids contacted by the protein and / or 5-10 or 2-5 or 1-3 amino acids outside of this region. In some examples, the epitope comprises a series of discontinuous amino acids that are positioned close to one another when the receptor or fragment thereof is folded, i.e., a “conformational epitope”. The skilled artisan will also be aware that the term "epitope" is not limited to peptides or polypeptides. For example, the term “epitope” includes chemically active surface groupings of molecules such as sugar side chains, phosphoryl side chains, or sulfonyl side chains, and, in certain examples, may have specific three-dimensional structural characteristics, and / or specific charge characteristics.

[0424] The term “plasma” shall refer to the straw-coloured / pale yellow component of blood obtained from one or more blood donor(s). Methods of obtaining plasma from a donor will be apparent to a skilled person and / or described herein. For example, plasma is obtained by removing red blood cells from donated blood. For example, plasma is obtained by plasmapheresis.

[0425] The term “plasma fraction” or “fraction thereof’ shall refer to plasma which has been fractionated to isolate one or more desirable protein components from the plasma. For example, plasma may be fractionated to isolate cryo-precipitates (proteins that precipitate out of solution when a unit of fresh frozen plasma is slowly thawed in the cold) and cryosupematant (also known as cryo-poor plasma). For example, plasma may be fractionated by ethanol precipitation to produce IgG-containing Oncley fractions, Cohn fractions, ammonium sulphate precipitates, or Precipitates A (KN A), B (KN B), and the Precipitate of Supernatant B (KN B+l) from plasma as described in US patent 3,301,842. Plasma fractions include II+III precipitate produced according to Cohn methods such as Method 6, Cohn et. al. J. Am; Chem. Soc., 68 (3), 459-475 (1946), Method 9, Oncley et al. J. Am; Chem. Soc., 71, 541-550 (1946), or the 1+11+111 precipitate, Method 10, Cohn et.al. J. Am; Chem. Soc., 72, 465-474 (1950); as well as the method of Deutsch et.al. J. Biol. Chem. 164, 109-118 (1946) or the Precipitate- A, B and the Precipitate of Supernatant B of Nitschmann and Kistler Vox Sang. 7, 414-424 (1962); Helv. Chim. Acta 37, 866-873 (1954). For example, the plasma may be fractionated by octanoic acid fractionation as described in European application 893450. Typically, Cohn Fractions, and Kistler / Nitschmann Precipitate’s A (KN A), B (KN B) and the Precipitate of Supernatant B (KN B+l) exist as a suspended paste. Other purification techniques including chromatography may be used.

[0426] The term “immunoglobulin G (IgG)”, also known as “gamma globulin” or “immune globulin”, shall be taken to mean antibody of isotype G. There are several subclasses of IgG, for example, IgGl, IgG2, IgG3 and IgG4.

[0427] The term “cryo-precipitate” or “cryo-precipitates” refers to proteins in plasma that precipitate out of solution when a unit of fresh frozen plasma is slowly thawed in the cold. Cryoprecipitates include factor VIII, fibrinogen, von Willebrand factor, factor XIII and platelet membrane microparticles.

[0428] The term “cryo-poor plasma” shall be taken to mean plasma removed of cryo-precipitates.

[0429] The term “cryo-rich plasma” shall be taken to mean plasma comprising components typically found in cryo-precipitates.

[0430] The term “partially purified plasma or plasma fraction thereof’ shall be understood to mean any fraction or solution from any stage of plasma purification.

[0431] The term “subject” shall be taken to mean any animal including humans, for example a mammal. Exemplary subjects include but are not limited to humans and non-human primates. For example, the subject is a human.

[0432] Detecting, identifying, and screening for ligands capable of inducing oligomerization and / or crosslinking of the ligand’s receptor

[0433] The present disclosure provides a method of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the ligand’s receptor, the method comprising:

[0434] (i) contacting the sample comprising the target ligand to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the target ligand in the sample to thereby form an immunocomplex; (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0435] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0436] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the target ligand in the sample.

[0437] Binding moieties

[0438] The present disclosure provides a binding moiety comprising a receptor, fragment or epitope thereof that specifically binds to a binding domain of the target ligand.

[0439] As used herein, the term “binding moiety” refers to any molecule, or part thereof, that is capable of specifically binding to a binding domain of the target ligand. As described herein, the binding moiety comprises a receptor, fragment or epitope thereof that specifically binds to a binding domain of the target ligand.

[0440] As used herein, the term “specifically binds” or “binds specifically” shall be taken to mean that a receptor, fragment or epitope of the disclosure reacts or associates more frequently, more rapidly, with greater duration and / or with greater affinity with a particular ligand (i.e., a binding domain of the target ligand) than it does with alternative ligands. For example, a receptor, fragment or epitope binds to a binding domain with materially greater affinity (e.g., 20 fold or 40 fold or 60 fold or 80 fold to 100 fold or 150 fold or 200 fold greater affinity) avidity, more readily, and / or with greater duration than it binds to other binding domains. Generally, but not necessarily, reference to binding means specific binding, and each term shall be understood to provide explicit support for the other term.

[0441] In one example, the first and second binding moieties specifically bind to the target ligand in the sample to thereby form an immunocomplex.

[0442] As used herein, the term “immunocomplex” shall be understood to refer to a molecule formed from the binding of the first and second binding moieties to the target ligand. It will be apparent to the skilled person that binding of the binding moieties to the target ligand result in a complex that acts as a unitary molecule.

[0443] Receptors, fragments or epitopes thereof

[0444] The present disclosure provides a binding moiety comprising a receptor, fragment or epitope thereof that specifically binds to a binding domain of the target ligand. For example, the present disclosure provides a first binding moiety comprising a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand and a second binding moiety comprising a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand.

[0445] It will be apparent to the skilled person from the disclosure herein, that receptors, fragments or epitopes thereof suitable for use in the present disclosure are dimerized, oligomerised and / or cross-linked upon binding of the target ligand to their receptor. As used herein, the term “oligomerization” in relation to the receptor, fragment or epitope thereof refers to an arrangement of monomeric units of the receptor into homo- or heterooligomers.

[0446] As used herein, the term “cross-linking” in relation to the receptor, fragment or epitope thereof refers to the process of chemically joining two or more receptor molecules by a covalent bond.

[0447] The skilled person will be aware of suitable receptors, fragments or epitopes thereof that oligomerize and / or cross link upon binding of their respective ligand and that are suitable for use in the present disclosure.

[0448] Exemplary receptors, fragments or epitopes thereof suitable for use in the present disclosure include a Fc receptor, fragment or epitope thereof; a T cell receptor, fragment or epitope thereof; a B cell receptor, fragment or epitope thereof; an epidermal growth factor (EGF) receptor, fragment or epitope thereof; a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof; an insulin receptor, fragment or epitope thereof; a programmed death 1 (PD-1) receptor, fragment or epitope thereof; and combinations thereof.

[0449] In one example, the first and / or second receptor, fragment or epitope thereof is a fragment crystallizable (Fc) receptor (FcR), fragment or epitope thereof.

[0450] The skilled person will understand that a FcR binds a Fc domain of a human immunoglobulin. However, the FcR may bind an immunoglobulin of any other mammalian species, including for example, a Camelid species, a rodent (e.g. a mouse, rat, rabbit, guinea pig) or non-human primate (e.g. chimpanzee, macaque) species. It will be appreciated that the scope of the present disclosure encompasses alleles, variants and mutations of FcR.

[0451] The skilled person will appreciate that there are several different types of FcRs based on the type of antibody they recognise. For example, those that bind IgG, are called Fc-gamma receptors (FcyR), those that bind IgA are called Fc-alpha receptors (FcaR) and those that bind IgE are called Fc-epsilon receptors (FcaR).

[0452] In one example, the FcR is a Fc-gamma receptor (FcyR). For example, the FcyR is a FcyRI (CD64), a FcyRIIA (CD32), a FcyRIIB (CD32), a FcyRIIIA (CD16a), and / or a FcyRIIIB (CD16b) The skilled person will understand that oligomerization of FcyR occurs when antibodies bind to Fey receptors on immune cells like macrophages and neutrophils, typically during antibody-dependent cellular cytotoxicity (ADCC). Cross-linking of FcyRs triggers receptor clustering and signal initiation. It will be apparent to the skilled person that oligomerization of FcyR induces activation of tyrosine-protein kinase, leading to activation of the PI3K, MAPK, and NF-KB pathways. This induces phagocytosis, cytokine production, and oxidative burst responses.

[0453] In one example, the FcR is a Fc-alpha receptor (FcaR). For example, the FcaR is a FcaRI (CD89).

[0454] In one example, the FcR is a Fc-epsilon receptor (FcaR). For example, the FcaR is a FcaRI or a FcaRII (CD23).

[0455] As used herein, the term “Fc epsilon Receptor I (FcaRI)” shall be understood to refer to an antigen present on mast cells, basophils, epidermal Langerhans cells, eosinophils and other antigen-presenting cells. FcsRI is multimeric receptor and a member of a family of related antigen / Fc receptors which have conserved structural features and similar roles in initiating intracellular signalling cascades. In humans, two types of FcsRI are expressed on the cell surface, a tetrameric receptor consisting of an a-chain, a P-chain and a homodimeric y-chain (aPy2) and a trimeric receptor consisting of an a-chain and a homodimeric y-chain (ayi).

[0456] In one example, the FcsRI is an a-chain of FcsRI or subunit thereof, or a P-chain of FcsRI or subunit thereof.

[0457] FcsRI is responsible for the activation and degranulation of mast cells and basophils. Antigenic crosslinking for example, through IgE and / or IgG binding, of the FcsRI initiates multiple signalling pathways, referred to as “FcsRI mediated signalling pathway”, which control diverse effector responses. These include the secretion of allergic mediators and induction of cytokine gene transcription, resulting in secretion of molecules such as interleukin-4, interleukin-6, tumour-necrosis factor-a and granulocyte-macrophage colony-stimulating factor. FcsRI is therefore central to the induction and maintenance of an allergic response.

[0458] In one example, the FcR is a polymeric immunoglobulin receptor (plgR). The skilled person will be aware that a plgR recognized the J chain region of polymerized IgA and IgM.

[0459] In one example, the first and / or second receptor, fragment or epitope thereof is a T cell receptor, fragment or epitope thereof.

[0460] The skilled person will understand that oligomerization of T cell receptors is induced by antigen recognition through the presentation of peptide-MHC complexes by antigen-presenting cells (APCs). T cell receptor engagement leads to clustering of receptors and recruitment of coreceptors like CD4 or CD8, which bring tyrosin-protein kinase into proximity with the T cell receptor complex. Oligomerization of the T cell receptors induces activation of ZAP-70 kinase, leading to the phosphorylation of linker for activated T cells (LAT), which then forms a scaffold for the assembly of signaling complexes that activate the Ras-MAPK, NF-KB, and calciumdependent pathways.

[0461] In one example, the T cell receptor comprises a T cell receptor alpha (a) chain and a T cell receptor beta (P) chain. In one example, the T cell receptor comprises a T cell receptor gamma (y) chain and a T cell receptor delta (5) chain. In one example, the T cell receptor chains are complexed with invariant CD3 molecules. In one example, the CD3 molecules comprise CD3(^, CD3y, CD35 and / or CD3s. In one example, the T cell receptor complex comprises T cell receptor a-P-CD3sy- CD3sb-CD3(X

[0462] In one example, the first and / or second receptor, fragment or epitope thereof is a B cell receptor, fragment or epitope thereof. The skilled person will recognise that the B cell receptor is composed of a membrane-bound immunoglobulin molecule linked to a CD79A (Iga) and CD79B (IgP) signal transduction moiety. Binding of antigens to B cell receptors on B cells induces receptor clustering and oligomerization. This clustering recruits and activates kinases such as tyrosine protein kinase, which phosphorylates immunoreceptor tyrosine-based activation motif (ITAMs) on the Iga and IgP subunits of the B cell receptor complex. Activation of tyrosine kinase leads to the activation of various pathways, including the PI3K / Akt, Ras-MAPK, and NF-KB pathways, promoting B cell activation and differentiation.

[0463] In one example, the first and / or second receptor, fragment or epitope thereof is an epidermal growth factor (EGF) receptor, fragment or epitope thereof. The skilled person will recognise that ligand binding (e.g., EGF) induces dimerization (homo- or heterodimerization) of EGF receptor molecules, leading to the autophosphorylation of the intracellular tyrosine kinase domain. Phosphorylation sites on EGF receptor serve as docking sites for signalling molecules, activating pathways such as Ras-MAPK, PI3K / Akt, and JAK / STAT, which are involved in cell growth, proliferation, and survival.

[0464] In one example, the first and / or second receptor, fragment or epitope thereof is a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof. The skilled person will recognise that reference to a “TNF receptor superfamily, fragment or epitope thereof’ refers to a protein superfamily of cytokine receptors characterized by the ability to bind TNFs via an extracellular cysteine-rich domain. For example, the TNF receptor superfamily is a TNFR1, a TNFR2, lymphotoxin beta receptor, 0X40, cluster of differentiation (CD) 40 (CD40), Fas receptor, a decoy receptor, CD27, CD30, 4-1BB, a death receptor, receptor activator of nuclear factor K B (RANK), osteoprotegerin, TNF-related weak inducer of apoptosis (TWEAK) receptor, transmembrane activator and CAML interactor (TACI), B-cell activating factor (BAFF) receptor and / or herpesvirus entry mediator. In their active form, the majority of TNF receptors form trimeric complexes in the plasma membrane. Trimerization of TNF receptors by binding of ligands such as TNF-a or CD40L leads to the recruitment of adaptor proteins like TNFR1 -associated death domain protein (TRADD), TNF receptor associated factors (TRAFs), and receptor interacting serine / threonine protein kinase 1 (RIPK1).

[0465] In one example, the first and / or second receptor, fragment or epitope thereof is an insulin receptor, fragment or epitope thereof. The skilled person will recognise that an insulin receptor is a transmembrane receptor that is activated by insulin and insulin growth factor (IGF-I, IGF-II) and belongs to the large class of receptor tyrosine kinase. Insulin binding induces dimerization of the receptor, leading to autophosphorylation of the tyrosine kinase domains. Phosphorylation of insulin receptor substrates recruits PI3K, which activates Akt, leading to glucose uptake, glycogen synthesis, and other metabolic effects.

[0466] In one example, the first and / or second receptor, fragment or epitope thereof is a programmed death 1 (PD-1) receptor, fragment or epitope thereof. The skilled person will recognise that PD-1 receptor is also known as cluster of differentiation 279 (CD279) and is a cell surface receptor on T and B cells. PD-1 oligomerizes when its ligands, PD-L1 or PD-L2, bind to it on T cells. This induces inhibitory signals that dampen T cell activation. Oligomerization induces recruitment of SHP-2 phosphatase, which dephosphorylates signalling molecules downstream of the TCR, such as ZAP-70, leading to inhibition of T cell proliferation and cytokine production.

[0467] Nucleic Acid Labels The present disclosure provides a binding moiety comprising a nucleic acid label. For example, the present disclosure provides a first binding moiety comprising a first nucleic label and a second binding moiety comprising a second nucleic acid label.

[0468] As used herein, the term “label” refers to a molecule that facilitates identification of the immunocomplex. For example, based on proximity between the first nucleic acid label and the second nucleic acid label a nucleic acid reporter from the immunocomplex is generated. Accordingly, it will be apparent to the skilled person that recognition of a ligand by the first and second binding moieties (each comprising a nucleic acid label) triggers the formation of amplifiable products.

[0469] In one example, the nucleic acid reporter is formed by proximity ligation or proximity extension.

[0470] As used herein, “proximity ligation” refers to when during formation of the immunocomplex, the first nucleic acid label and the second nucleic acid label are brought into sufficient proximity to be ligated, and a fragment of the ligation product is used as an amplicon to generate the signal for detection.

[0471] As used herein, “proximity extension” refers to when during formation of the immunocomplex, the first nucleic acid label and the second nucleic acid label are brought into sufficient proximity to interact with each other and form a duplex, such that the 3 ' end of at least one nucleic acid label of the duplex can be extended to generate an extension product, which can be used as an amplicon to generate the signal for detection.

[0472] The skilled person will recognise from the disclosure herein that the methods of the disclosure permit detection of multiple target ligands from multiple samples simultaneously.

[0473] In one example, the nucleic acid label further comprises a receptor barcode specific to the receptor, fragment or epitope thereof. For example, the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment or epitope thereof; and / or the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment or epitope thereof.

[0474] In one example, the nucleic acid label further comprises a sample barcode. For example, the first nucleic acid label and / or the second nucleic acid label comprises a sample barcode.

[0475] As used herein, reference to a “barcode” in relation to the sample or receptor refers to an identification sequence that can be incorporated into the nucleic acid label and decoded by DNA sequence or other methods. It will be apparent to the skilled person, that incorporation of the sample and / or receptor barcodes allows multiplexing of samples in a method of the disclosure.

[0476] In one example, the nucleic acid reporter comprises the first receptor barcode, the second receptor barcode and the sample barcode.

[0477] In one example, presence of the receptor barcode allows multiplexing to enable detection and measurement of multiple different target ligands in the same sample in parallel. For example, the assay methods provided herein simultaneously detect at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least twelve, at least fifteen, at least twenty, at least thirty, at least forty, at least fifty, at least sixty, at least seventy, at least eighty, at least ninety, or at least one hundred target ligands in the sample by simultaneously detecting the unique receptor barcode associated with each target ligand.

[0478] In one example, presence of the sample barcode allows multiplexing to enable detection and measurement of the same target ligand in a plurality of samples in parallel. For example, the assay methods provided herein simultaneously detect a target ligand in at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least twelve, at least fifteen, at least twenty, at least thirty, at least forty, at least fifty, at least sixty, at least seventy, at least eighty, at least ninety, or at least one hundred samples by simultaneously detecting the unique sample barcode in the nucleic acid reporter associated with each subject.

[0479] Methods of detecting and quantifying the nucleic acid reporter will be apparent to the skilled person and / or described herein. For example, the method comprises detecting the nucleic acid reporter by polymerase chain reaction (PCR), next generation sequencing, Rolling Cycle Amplification (RCA), strand displacement amplification (SDA), Loop-Mediated Isothermal Amplification (LAMP), Recombinase Polymerase Amplification (RPA), or a QuantiGene assay.

[0480] Nucleic Acid Tags and Capture Probes

[0481] The present disclosure provides a binding moiety comprising a nucleic acid tag comprising a biotinylated tail.

[0482] The present disclosure also provides a binding moiety comprising a nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence.

[0483] The present disclosure further provides a first binding moiety comprising a first nucleic acid tag comprising a biotinylated tail and a second binding moiety comprising a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence.

[0484] In one example, the nucleic acid tag binds the capture probe.

[0485] The skilled person will recognise that the nucleic acid tag and the capture probe can be coupled by virtue of covalent linkage (e.g., chemical cross-linking), by non-covalent association, nucleic acid binding or by hybridization.

[0486] In one example, the first nucleic acid tag comprises a biotinylated tail. For example, the first nucleic acid tag comprises a biotinylated tail that binds to the first capture probe.

[0487] In one example, the first nucleic acid tag comprises a biotinylated tail that binds to the first capture probe comprising a biotin binding protein. For example, the biotin binding protein is streptavidin or avidin.

[0488] In one example, the first nucleic acid tag comprises a biotinylated tail that binds to the streptavidin or avidin of the first capture probe. For example, the tag and probe couple by non- covalent association.

[0489] Suitable first nucleic acid tag-first capture probe binding pairs will be apparent to the skilled person and / or described herein. For example, fluorescein and anti-fluorescein, digioxigenin and anti-digioxigenin, and DNP (dinitrophenol) / anti-DNP. In one example, the second nucleic acid comprises an A, a T, a G and / or a C rich tail sequence. For example, the second nucleic acid comprises an A, a T, a G and / or a C rich tail sequence that binds to the second capture probe.

[0490] In one example, the second nucleic acid comprises an A, a T, a G and / or a C rich tail sequence that binds to the second capture probe comprising a complementary T, A, C and / or G rich sequence. For example, the second nucleic acid comprises an A rich tail sequence that binds to the second capture probe comprising a complementary T rich sequence. In another example, the second nucleic acid comprises a T rich tail sequence that binds to the second capture probe comprising a complementary A rich sequence. In a further example, the second nucleic acid comprises a G rich tail sequence that binds to the second capture probe comprising a complementary C rich sequence. In one example, the second nucleic acid comprises a C rich tail sequence that binds to the second capture probe comprising a complementary G rich sequence.

[0491] In one example, the A rich sequence is a poly-A sequence.

[0492] In one example, the T rich sequence is a poly-T sequence.

[0493] In one example, the G rich sequence is a poly-G sequence.

[0494] In one example, the C rich sequence is a poly-C sequence.

[0495] In one example, the nucleic acid tag binds the capture probe through nucleic acid hybridization. It will be apparent to the skilled person that dissociation of the hybridization link may be achieved, for example, through the change of salt concentration in buffer and / or increasing the temperature of the reaction, without affecting the integrity of the immunocomplex. Furthermore, capture / release through hybridization can be renewed and repeated multiple cycles without deterioration of efficiency and selectivity.

[0496] It will be apparent to the skilled person that the A, T, G and / or C rich sequence has a short length such that the binding from the complementary T, A, C and / or C sequence on the second capture probe is weaker than binding between the first and second binding moieties and the target ligand, thus keeping immunocomplex stable in any releasing steps.

[0497] The present disclosure provides contacting the immunocomplex to a first and / or second capture probe immobilized on a first and / or second solid matrix, wherein the first and / or second capture probe binds to the first and / or second nucleic acid tag of the immunocomplex.

[0498] The present disclosure provides contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag of the immunocomplex.

[0499] The present disclosure further provides contacting the immunocomplex to a second capture probe immobilized on a second solid matrix, wherein the second capture probe binds the second nucleic acid tag of the immunocomplex.

[0500] The term “matrix” in the context of the “solid matrix” shall be taken to mean a support to which molecules may be attached, directly or indirectly. The matrix may include any substrate material that is capable of providing physical support for the compositions described herein. The materials may be naturally occurring, synthetic, or a modification of a naturally occurring material. Suitable matrix materials may include glass fibers, polyester, cellulose, rayon, silicon, a silicon wafer chip, graphite, mirrored surfaces, laminates, membranes, ceramics, plastics (including polymers such as, e.g., poly(vinyl chloride), cyclo-olefin copolymers, agarose gels or beads, polyacrylamide, polyacrylate, polyethylene, polypropylene, poly(4-methylbutene), polystyrene, polymethacrylate, poly(ethylene terephthalate), polytetrafluoroethylene (PTFE or Teflon®), nylon, poly(vinyl butyrate)), germanium, gallium arsenide, gold, silver, Langmuir Blodgett films, a flow through chip, etc., either used by themselves or in conjunction with other materials. Additional rigid materials may be considered, such as glass, which includes silica and further includes, for example, glass that is available as Bioglass. Other materials that may be employed include porous materials, such as, for example, controlled pore glass beads, crosslinked beaded Sepharose® or agarose resins, or copolymers of crosslinked bis-acrylamide and azalactone.

[0501] In one example, the method further comprises washing the solid matrix to remove unbound immunocomplex. For example, the method comprises washing the first and / or second solid matrix to remove unbound immunocomplex.

[0502] In one example, the method further comprises releasing the bound immunocomplex from the immobilized capture probe. For example, the method comprises releasing the bound immunocomplex from the second immobilized capture probe.

[0503] It will be apparent to the skilled person that the multiple rounds of capture and release reduce any non-specific background signal, and any further background signal may be reduced by the addition of further rounds of capture and release.

[0504] Methods of achieving releasable bonds between the nucleic acid tag and the capture probe will be apparent to the skilled person and / or described herein. Exemplary releasable bonds include via a thioester group, a disulfide linkage, a cleavable linkage or a protein-protein interaction.

[0505] In one example, the releasable bond is a linkage that can be cleaved with appropriate enzymatic activities, including for example, phosphodiester , phospholipid, ester or b-galactose.

[0506] In one example, the releasable bond is a linkage that can be cleaved by chemoenzymatic reactions, such as Staphy-eSrtA pair.

[0507] In one example, the releasable bond is formed between arginine residues and a sorbent derivatized with 4-(oxoacetyl)phenoxyacetic acid.

[0508] In one example, the releasable bond is a non-covalent bond (e.g., a hydrogen bond) disrupted through binding competition.

[0509] Target Ligands

[0510] The present disclosure provides a method of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the ligand’s receptor.

[0511] As used herein, the term “target ligand” refers to any substance or molecule to be detected by the methods provided herein.

[0512] Target ligands suitable for use in the present disclosure will be apparent to the skilled person and / or described herein. In one example, the target ligand is an immunoglobulin (Ig), a peptide-MHC complex and / or an antigen.

[0513] The term “immunoglobulin (Ig)” shall be taken to mean a protein or antibody present in the serum and cells of the immune system. There are several types of Ig, for example, IgG, IgE, IgM, IgD, IgA, and IgY. The term “immunoglobulin G (IgG)”, also known as “gamma globulin” or “immune globulin”, shall be taken to mean antibody of isotype G. There are several subclasses of IgG, for example, IgGl, IgG2, IgG3 and IgG4.

[0514] The skilled artisan will be aware that an “antibody” is generally considered to be a protein that comprises a variable region made up of a plurality of polypeptide chains, e.g., a polypeptide comprising a light chain variable region (VL) and a polypeptide comprising a heavy chain variable region (VH). An antibody also generally comprises constant domains, some of which can be arranged into a constant region, which includes a constant fragment or fragment crystallizable (Fc), in the case of a heavy chain. A VH and a VL interact to form a Fv comprising an antigen binding region that is capable of specifically binding to one or a few closely related antigens. Generally, a light chain from mammals is either a K light chain or a light chain and a heavy chain from mammals is a, 5, a, y, or p. Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGi, IgG2, IgGs, IgG4, IgAi and IgA2) or subclass. The term “antibody” also encompasses humanized antibodies, primatized antibodies, human antibodies, synhumanized antibodies and chimeric antibodies.

[0515] The present disclosure provides a binding moiety comprising a receptor, fragment or epitope thereof that specifically binds to a binding domain of the target ligand.

[0516] As used herein, the term “binding domain” shall be taken to mean a structure formed by the target ligand that is capable of binding or specifically binding to an antigen (i.e., a receptor, fragment or epitope thereof). The binding domain need not be a series of contiguous amino acids, or even amino acids in a single polypeptide chain. For example, in a Fv produced from two different polypeptide chains the binding domain is made up of a series of amino acids of a VL and a VH that interact with the antigen (i.e., the receptor, fragment or epitope thereof) and that are generally, however not always in the one or more of the CDRs in each variable region. In some examples, the binding domain is a VH or a VL or a Fv.

[0517] As used herein, “variable region" refers to the portions of the light and / or heavy chains of an antibody as defined herein that is capable of specifically binding to an antigen and includes amino acid sequences of complementarity determining regions (CDRs); i.e., CDR1, CDR2, and CDR3, and framework regions (FRs). Exemplary variable regions comprise three or four FRs (e.g., FR1, FR2, FR3 and optionally FR4) together with three CDRs. VH refers to the variable region of the heavy chain. VL refers to the variable region of the light chain.

[0518] In one example, the immunoglobulin is an anti-Fc receptor Ig. For example, an anti-FcyR Ig, an anti-FcaR Ig or an anti-FcsR Ig. In one example, the anti-FcsR Ig is an anti-FcsRI Ig or an anti-Fc8R.II Ig.

[0519] The present disclosure provides a method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample. The present disclosure also provides a method of screening a sample for an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig).

[0520] The present disclosure further provides a method of identifying a sample suitable for administration to a subject, wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0521] In one example, the target ligand is an antigen. For example, the antigen is selected from the group consisting of an EGF, a programmed death ligand 1 (PD-L1), a PD-L2, a TNF-a, a TNF- C, an OX40L, a CD154, a FasL, a CD70, a Siva, a CD153, a 4-1BB ligand, a TNF-related apoptosis inducing ligand (TRAIL), a RANK ligand (RANKL), TWEAK, a proliferation inducing ligand (APRIL), BAFF, a calcium modulating ligand (CAMLG), LIGHT, TNF-like ligand 1A (TL1A) and combinations thereof.

[0522] Analysis of effector cells

[0523] Methods of determining the proportion of basophil and / or mast cell activation and / or basophil and / or mast cell degranulation of the disclosure will be apparent to the skilled person and / or described herein. For example, the proportion can be determined using ELISA, FLISA or a lateral flow assay. Briefly, basophil and / or mast cells are incubated with the plasma sample or fraction thereof for a suitable period time. Cells are then fixed and the proportion of basophil and / or mast cell activation and / or basophil and / or mast cell degranulation relative to the total proportion of basophil and / mast cells being exposed to the sample is analysed using flow cytometry.

[0524] As used herein, the term “proportion” in reference to a basophil and / or mast cell activation shall be understood to refer to a measure of an activation marker.

[0525] As used herein, the term “activation” shall be understood to mean the stimulation of a cell (e.g., basophils and / or mast cells) by exposure to activation markers, such as a cellular or soluble ligand, which results in a change in the morphology or behaviour of the cell.

[0526] For example, upon activation of the cell, the cell upregulates the activation markers. In some examples, the activation markers are selected from the group consisting of CD63, CD203c, CD 107a and combinations thereof.

[0527] As used herein the term “degranulation” shall be taken to mean the process by which cytoplasmic granules are released from cells (e.g., of mast cells and / or basophils).

[0528] In some examples, activation of the cell leads to degranulation. In other examples, activation of the cell does not lead to degranulation. For example, activation of the cell leads to increased cytokine production without degranulation.

[0529] In one example, basophil activation following exposure to the plasma sample or fraction thereof can be determined using basophil activation tests known in the art. For example, the test is a basophil activation test. The test uses activation markers to measure the proportion of activation and / or degranulation of basophils. For example, the activation marker is CD63. In another example, the activation marker is CD203c. CD63 and CD203c are useful markers for flow cytometric quantification of in vitro activated basophils. In some examples, the basophils are activated human basophils.

[0530] In one example, the proportion of basophil activation is determined using flow cytometry. In one example, the proportion of basophil activation is determined by incubating basophils with a plasma sample or fraction thereof and quantifying the proportion of CD63 expressing basophils in the total population of basophils using flow cytometry. In another example, the proportion of basophil activation is determined by incubating basophils with a plasma sample or fraction thereof and quantifying the proportion of CD203c expressing basophils in the total population of basophils using flow cytometry.

[0531] In one example, basophil activation is measured by the presence of interleuklin-3 (IL-3). In another example, basophil activation is measured by the absence of IL-3.

[0532] In another example, mast cell activation following exposure to the plasma sample or fraction thereof can be determined using mast cell activation assays known in the art. For example, the assay is a Hoxb8 mast cell activation test. The assay uses activation markers to measure the proportion of activation and / or degranulation of mast cells. For example, the activation marker is CD 107a. CD 107a is a useful marker flow cytometric quantification of in vitro activated mast cells. In some examples, the mast cells are Hoxb8 mast cells.

[0533] In one example, the proportion of mast cell activation is determined by incubating mast cells with a plasma sample or fraction thereof and quantifying the proportion of CD107a expressing mast cells in the total population of mast cells using flow cytometry.

[0534] Briefly, the basophils or mast cells are incubated with the plasma sample or fraction thereof. Detecting activation of the cells is done by detecting the proportion of cells expressing an activation marker (e.g., CD63, CD203c and / or CD107a). The proportion of activation (i.e., the proportion of cells expressing the activation marker in the total population of cells) is then quantified using flow cytometry.

[0535] It will be apparent to the skilled person from the disclosure herein that methods of determining basophil and / or mast cell activation and / or degranulation are performed in vitro.

[0536] In one example, the method comprises determining a detectable amount of anti-FcsRI Ig in the plasma sample or fraction thereof by determining the proportion of basophils expressing CD63 following exposure to the plasma sample or fraction thereof. In one example, a detectable amount of anti-FcsRI Ig is present in the plasma sample or fraction thereof if at least 5% of a population of basophils express CD63 following exposure to the plasma sample or fraction thereof. For example, the proportion of basophils expressing CD63 following exposure to the plasma sample or fraction thereof is more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 11%, more than 12%, more than 13%, more than 14% or more than 15% of the total population of basophils exposed to the plasma sample or fraction thereof.

[0537] In one example, the method comprises determining a detectable amount of anti-FcsRI Ig in the plasma sample or fraction thereof by determining the proportion of basophils expressing CD203c following exposure to the plasma sample or fraction thereof. In one example, a detectable amount of anti-FcsRI Ig is present in the plasma sample or fraction thereof if at least 5% of a population of basophils express CD203c following exposure to the plasma sample or fraction thereof. For example, the proportion of basophils expressing CD203c following exposure to the plasma sample or fraction thereof is more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 11%, more than 12%, more than 13%, more than 14% or more than 15% of the total population of basophils exposed to the plasma sample or fraction thereof.

[0538] In one example, the method comprises determining a detectable amount of anti-FcsRI Ig in the plasma sample or fraction thereof by determining the proportion of mast cells expressing CD 107a following exposure to the plasma sample or fraction thereof. In one example, a detectable amount of anti-FcsRI Ig is present in the plasma sample or fraction thereof if at least 5% of a population of mast cells express CD 107a following exposure to the plasma sample or fraction thereof. For example, the proportion of mast cells expressing CD 107a following exposure to the plasma sample or fraction thereof is more than 5%, more than 6%, more than 7%, more than 8%, more than 9%, more than 10%, more than 11%, more than 12%, more than 13%, more than 14% or more than 15% of the total population of mast cells exposed to the plasma sample or fraction thereof.

[0539] Methods of Preparing anti-FcsRI Ig Reduced Preparations

[0540] The present disclosure provides a method of preparing an anti-FcsRI Ig reduced preparation from a plasma or a fraction thereof using a chromatography resin. For example, the disclosure provides a method of preparing an anti-FcsRI Ig reduced preparation from a plasma or a fraction thereof using an affinity chromatography resin. The affinity resin of the present disclosure comprises a ligand capable of specifically binding to an anti-FcsRI Ig.

[0541] The present disclosure further comprises an affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig immobilized to a matrix of the affinity chromatography resin. In one example, the resin further comprises a blood group A antigen and a blood group B antigen immobilized to the matrix of the affinity chromatography resin.

[0542] As used in this context, “a ligand capable of specifically binding to an anti-FcsRI Ig” or “a ligand which binds to the anti-FcsRI Ig” shall be understood to mean a molecule immobilised to a matrix of the chromatography column which interacts anti-FcsRI Ig. For example, the ligand comprises a FcsRI or fragment or epitope thereof. In one example, the ligand is a FcsRI or fragment or epitope thereof. In another example, the ligand is a FcsRI antibody or fragment thereof. In a further example, the ligand is a FcsRI DNA aptamer.

[0543] The present disclosure provides a method comprising binding the anti-FcsRI Ig to a chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the sample and collecting the anti-FcsRI Ig reduced preparation.

[0544] In one example, the method of detecting of the disclosure is performed to detect if anti- FcsRI Ig is present in the sample before the sample is loaded onto the chromatography resin. In one example, the method of screening of the disclosure is performed to screen if anti-FcsRI Ig is present in the sample before the sample is loaded onto the chromatography resin. In one example, the method of identifying of the disclosure is performed to identify if anti-FcsRI Ig is present in the sample before the sample is loaded onto the chromatography resin.

[0545] The methods of detecting, screening and identifying of the disclosure also comprise preparing an anti-FcsRI Ig reduced preparation from the sample if anti-FcsRI Ig is present in the sample, using a chromatography resin. For example, the disclosure provides a method of preparing an anti-FcsRI Ig reduced preparation from a plasma or a fraction thereof using an affinity chromatography resin.

[0546] The affinity resin of the present disclosure comprises a ligand capable of specifically binding to an anti-FcsRI Ig.

[0547] The term “affinity chromatography resin” shall be taken to mean a resin comprising an affinity chromatography ligand (e.g. FcsRI, or fragment or epitope thereof) attached to a matrix as would be apparent to the skilled artisan and / or as described herein.

[0548] As used herein, the terms “reduced” and “depleted” are used interchangeably and shall be understood to mean an Ig preparation wherein the amount of anti-FcsRI Ig has been decreased relative to an Ig preparation where the anti-FcsRI Ig has not been reduced or depleted and in an amount sufficient to prevent activation of the FcsRI mediated signalling pathway. As would be appreciated by the skilled person by the disclosure herein, the anti-FcsRI Ig need not be reduced or depleted 100% but only in in an amount to prevent activation of the FcsRI mediated signalling pathway.

[0549] As used herein, the terms “reduced basophil activation” and “reduced mast cell activation” shall be understood to mean an Ig preparation where the amount of anti-FcsRI Ig is reduced such that the activation of the basophil or mast cell is decreased. As would be appreciated by the skilled person reduced basophil activation or reduced mast cell activation need not refer to 100% decrease in the activation, but merely a decrease relative to an Ig preparation where the anti-FcsRI Ig has not been reduced or depleted. Suitable affinity chromatography resins will be apparent to the skilled person and / or described herein. In one example, the resin comprises a ligand comprising a FcsRI or fragment or epitope thereof. In another example, the resin comprises a ligand comprising a FcsRI antibody or fragment thereof. The skilled person will be aware that ligands based on FcsRI are capable of specifically binding to all types of anti-FcsRI Ig (i.e., IgG and IgE).

[0550] Continuous affinity chromatography

[0551] In some examples, the affinity chromatography is continuous affinity chromatography.

[0552] The term “continuous affinity chromatography” shall be taken to mean a chromatographic method comprising one or more column(s) packed with identical affinity resins, wherein each column comprises one or more zones. A zone is a column, or a region of a column, comprising the resin where one or more chromatography steps can be performed. For example, a zone is selected from a group consisting of an equilibration zone, a binding zone, a wash zone, an elution zone, a stripping zone, or a combination thereof. Continuous affinity chromatography comprising more than one column involves the columns being connected in an arrangement that allows the columns to be operated in series and / or in parallel. In principle, IgG may be loaded on a first and / or subsequent columns while other columns (or other zones of a column) are going through an equilibration, wash, elution, and / or regeneration simultaneously. Examples of continuous affinity chromatography will be apparent to the skilled person and / or described herein.

[0553] Examples of columns which may be used to perform the continuous chromatography method will be apparent to the skilled person and / or described herein. For example, the continuous chromatography method may be performed using Tricorn 5 / 100 (Cytiva). In another example, the continuous chromatography method may be performed using BioSMB PD System (Sartorius).

[0554] Simulated moving bed (SMB) chromatography

[0555] In one example, the continuous affinity chromatography is simulated moving bed (SMB) chromatography. The term “simulated moving bed chromatography” or “SMB chromatography” refers to a chromatography method first described in US patent 2,985,589. Examples of SMB chromatography setup and / or apparatus will be apparent to the skilled person and / or described herein. The concept of simulated moving bed involves the use of multiple smaller columns (rather than one large column) containing a solid absorbent (e.g. affinity resin) and performing one or more continuous chromatography steps (i.e. equilibration, binding, washing, eluting or stripping) simultaneously on different columns in a continuous loop.

[0556] An example of a SMB chromatography set up has columns arraged into four sections with one or more columns per section. Two inlet streams (feed and eluent) and two outlet streams (extract and raffinate) are directed in alternating order to and from the column ring. The inlet and outlet positions are switched at regular time intervals in the direction of the liquid flow, thus simulating counter-current movement of columns. A feed (containing adsorbable components (extract)) is loaded onto one or more columns of the SMB chromatography setup, and the extract binds to the resin within the columns. Meanwhile, less adsorbed components (raffinate) in the feed pass through the column. The raffinate may be loaded onto one or more subsequent column(s) or removed from the SMB chromatography system as waste. An eluent is loaded onto the column to collect the extract. For example, an eluate may be collected from a first column while more feed is loaded onto one or more subsequent column(s).

[0557] Suitable wash and elution buffers having the characteristics of the present disclosure will be apparent to the skilled person and / or described herein. In one example, the wash buffer comprises 20 mM sodium dihydrogen phosphate, 145 mM sodium chloride and is at a pH of 7.4.

[0558] The resin in SMB chromatography may undergo multiple cycles (e.g. 50 cycles) of resin equilibration, IgG loading, binding, elution, stripping, sanitising, and / or regeneration per batch of plasma sample or fraction thereof used. Multiple batch runs (e.g. 4 to 10 batches) may be performed using SMB chromatography. The total life time of the resin in SMB chromatography can be in the range of 200 to 500 cycles (if not more) before the resin is unusable. Resin regeneration is generally performed to allow multiple uses of the resin. Periodic counter-current chromatography (PCC)

[0559] In one example, the continuous affinity chromatography is periodic counter-current chromatography (PCC). Examples of PCC setup and / or apparatus will be apparent to the skilled person and / or described herein. The concept of PCC involves the use of multiple columns containing a solid absorbent (e.g. affinity resin) and performing the chromatography steps in parallel in a quasi -continuous manner. The buffers used in binding, washing, and / or elution steps flow counter-current to the affinity resin.

[0560] An example of PCC setup involves the use of two columns. In a first step, a sample is loaded onto a first column above the DBC of the resin so that unbound product (e.g. IgG) breaks through the first column and is captured by the second column. In a second step, the first column is washed, eluted, cleaned and / or re-equilibrated independently of the second column being loaded with a further sample. In a third step, an additional sample is loaded onto the second column above the DBC of the resin so that unbound product breaks through the second column and is captured by the first column. In a fourth step, the second column is washed, eluted, cleaned and / or reequilibrated independently of the first column being loaded with a further sample. The process steps are continuously cycled between the two columns.

[0561] Another example of PCC setup involves the use of multiple columns. For example, a variation of the above PCC setup can involve use of multiple columns to capture unbound product which simulates use of a large column.

[0562] Continuous counter-current tangential chromatography (CCTC)

[0563] In one example, the continuous affinity chromatography is continuous counter-current tangential chromatography (CCTC). Examples of CCTC setup and / or apparatus will be apparent to the skilled person and / or described herein. The concept of CCTC involves using the affinity resin in a slurry form where the slurry is continuously directed through a number of static mixers and hollow fiber membranes which separate the fluid phase from the resin. CCTC is ordinarily performed at low pressures (e.g. <70kPa).

[0564] An example of a CCTC process involves binding, first wash, second wash, elution, stripping and / or equilibration steps. Sample (e.g. plasma sample or fraction thereof) and the affinity resin is passed through static mixers and hollow fiber membranes in a binding step. Impurities are removed in the flow through of the hollow fiber membranes in the washing step, while resin bound product (i.e. IgG) is retained by the membrane. The hollow fibres retain the resin and allow the product to flow through in the elution step. The resins are stripped and / or equilibrated and process repeated.

[0565] Continuous counter-current spiral chromatography (CCSC)

[0566] In one example, the continuous affinity chromatography is continuous counter-current spiral chromatography (CCSC). Examples of CCSC setup and / or apparatus will be apparent to the skilled person and / or described herein. The concept of CCSC involves the use of a compact rotating coil separation column mounted onto a centrifuge rotary frame. There are two separation column designs currently available: the spiral disk assembly and the spiral tube support assembly.

[0567] An exemplary CCSC process involves a coiled separation column revolving around a central axis of the centrifuge while it synchronously rotates about its own axis (at e.g., 1,000 to 1,200 rpm). A mobile phase can be passed through the centrifuge rotor without rotary seals, and a large amount of a stationary phase is retained while the two phases are mixed along the length of the column to produce a highly efficient solute separation.

[0568] Buffers

[0569] The present disclosure provides an affinity chromatography method using buffers which enable efficient Ig binding to, and collection from, the resin. Generally, plasma sample or fraction thereof are at a neutral pH (pH of about 7.4). The resin is equilibrated with an equilibration buffer and / or washed with a wash buffer having a buffering range which covers the neutral pH. Suitable wash buffers comprise buffering agents having a dissociation constant (pKa) between 6.8 and 8.5 at 25 °C.

[0570] An exemplary buffering agent of the equilibration and / or wash buffer is sodium dihydrogen phosphate, where the phosphoric acid component of sodium dihydrogen phosphate has three dissociation constants (pKa: 2.16, 7.21 and 12.32). Phosphoric acid has a dissociation constant at about the pH of an elution and / or stripping buffer used in the continuous affinity chromatography method. However, phosphoric acid does not have a dissociation constant between the pH of the equilibration and / or wash buffer (higher pH) and the elution and / or stripping buffer (lower pH) used in the affinity chromatography method. This enables a fast switch between wash and elution steps, and stripping and equilibration steps, giving more defined peaks and shorter chromatography phases. An advantage of using such equilibration and / or wash buffers is that smaller buffer volumes can be used, thereby increasing the efficiency of the affinity chromatography method.

[0571] Other suitable buffering agents of the equilibration and / or wash buffer include imidazole (pKa: 7.0), Tris (pKa: 8.30), glycylglycine (pKa: 8.40), MOPS (pKa: 7.2), PIPES (pKa: 6.8), TES (pKa: 7.40), Bicine (pKa: 8.35), HEPES (pKa: 7.55), EPPS (pKa:8.00), HEPPSO (pKa: 7.85), MOBS (pKa: 7.60), POPSO (pKa: 7.78), TAPSO (pKa: 7.61), Tricine (pKa: 8.05), TEA (pKa: 7.76).

[0572] Chromatography resin

[0573] The present disclosure provides a method preparing an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation from a plasma sample or a fraction thereof using a continuous chromatography resin. In one example, the resin of the present disclosure comprises a ligand capable of specifically binding to an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig).

[0574] In one example, the method further comprises use of an affinity chromatography resin to remove anti-A and anti-B antibodies. In one example, the method further comprises use of an affinity chromatography resin to remove anti-A antibodies. In one example, the method further comprises use of an affinity chromatography resin to remove anti-B antibodies.

[0575] Suitable chromatography resins will be apparent to the skilled person and / or described in US2009 / 074749 incorporated herein by reference. In one example, the resin comprises a mixture of supports whose matrixes are grafted with oligosaccharide groups which have antigenic similarity with blood groups A and B. An exemplary resin is Glycosorb Abo® (Glycorex Transplantation AS).

[0576] Analysis of the Anti-FcsRI Ig Reduced Preparation

[0577] In one example, the activity of the anti-FcsRI Ig reduced preparation is assessed. Methods of determining yield, purity and IgG subclass distribution will be apparent to the skilled person and / or described herein.

[0578] In one example, purity is determined by SDS-PAGE and MALDI-TOF-MS peptide fingerprint analysis. Briefly, purified plasma protein products or pharmaceutical compositions described herein are loaded onto a suitable SDS-PAGE gel (e.g. 8-16% TRIS-glycine), along with a protein size marker and a positive control for the protein of interest (e.g., IgG such as IVIG) under reduced and non-reduced conditions. Proteins are separated based on size and protein bands of interest are isolated, processed and analysed by MALDI-TOF-MS.

[0579] In another example, impurities in the anti-FcsRI Ig reduced preparation or pharmaceutical composition described herein are measured in an Enzyme-Linked Immunosorbent Assay (ELISA) using impurity (e.g. IgA) specific antibodies. For example, the ELISA is performed using commercially available methods.

[0580] In one example, purity, yield and / or subclass distribution of IgG is determined by nephelometry.

[0581] In one example, the anti-FcsRI Ig reduced preparation comprises a purity of more than 95% Ig. For example, the anti-FcsRI Ig reduced preparation comprises a purity of more than 96% Ig. In another example, the anti-FcsRI Ig reduced preparation comprises a purity of more than 97% Ig. In another example, the anti-FcsRI Ig reduced preparation comprises a purity of more than 98% Ig. In another example, the anti-FcsRI Ig reduced preparation comprises a purity of more than 99% Ig-

[0582] Stability of plasma and plasma fraction

[0583] The stability of the plasma sample or fraction thereof for loading onto a chromatography resin described herein can be determined by assessing the pro-coagulant activity, proteolytic activity and particle size of the plasma sample or fraction thereof. Methods for assessing procoagulant activity, proteolytic activity and particle size will be apparent to a skilled person and / or described herein. Briefly, the plasma or plasma fraction is freeze / thawed in one or more cycles, stored at between 2°C and 32°C (e.g. 2°C, 10°C, 18°C, 21 °C, 28°C or 32°C) for 24 or up to 48 hrs and analysed using one or more of the methods described below. In one example, the plasma sample or fraction thereof is thawed in one or more cycles at a temperature of 32°C, stored for 24 or up to 48 hours and analysed using one or more of the methods described below. In another example, the plasma sample or fraction thereof is thawed in one or more cycles at a temperature of 32°C, stored for 24 or up to 48 hours and analysed using one or more of the methods described below and then cooled and stored at a temperature of 21 °C. In one example, the plasma sample or fraction thereof is thawed at a temperature of 32°C and at a temperature of 21 °C before the continuous chromatography.

[0584] In one example, the pro-coagulant activity in the plasma sample or fraction thereof can be determined using an in vitro coagulation assay, e.g., activated partial thromboplastin time (NaPTT) assay. The NaPTT assay measures the rate at which one or more coagulation factors (e.g., fibrinogen, prothrombin, proaccelerin, anti-hemophilic factor, Stuart-Prower factor, plasma thromboplastin antecedent and Hegeman factor) are activated or form in plasma sample or fraction thereof, when coagulation activators (e.g. silica, kaolin, ellagic acid) are added to the assay.

[0585] In one example, proteolytic activity in the plasma sample or fraction thereof can be assessed by measuring the activity of thrombin, general serine proteases, kallikrein, plasmin and FXa e.g. using commercially available kits, such as thrombin activity assay kit (S-2238), general serine protease assay kit (S-2288), kallikrein activity assay kit (S-2302), plasmin activity assay kit (S- 2251) and FXa activity kit (S-2765).

[0586] In one example, the size of any particles in the plasma sample or fraction thereof is assessed by microflow imaging (MFI) and poly dispersity index is calculated. Calculation of the poly dispersity index will be apparent to the skilled person.

[0587] Pharmaceutical Compositions and Methods of Production thereof

[0588] The present disclosure provides methods of identifying a sample suitable for use in producing an immunoglobulin (Ig) preparation for administration to a subject.

[0589] The present disclosure further provides methods of screening a subject to determine suitability for plasma donation for production of an immunoglobulin (Ig) preparation. The present disclosure further provides methods of preparing an anti-FcsRI Ig reduced preparation from the sample if anti-FcsRI Ig is present in the sample, using a chromatography resin.

[0590] It will be apparent to the skilled person from the disclosure herein that the presence of the anti-FcsRI Ig in the sample indicates the donor is not suitable for plasma donation for the production of an Ig preparation due to activation the FcsRI mediated signaling pathway which induces mast cell activation and / or basophil activation.

[0591] In one example, the method is performed on a single plasma donation sample or fraction thereof. In another example, the method is performed on a pooled sample or fraction thereof (i.e., comprises more than one plasma donation sample).

[0592] As provided herein, a donor is suitable for plasma donation and / or a sample is suitable for use in the production of an Ig preparation for administration to a subject if a detectable amount of anti-FcsRI Ig is not present. For example, anti-FcsRI Ig in the sample at a concentration of less than 500 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 450 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 400 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 350 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 300 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 250 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 200 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation. In another example, anti-FcsRI Ig in the sample at a concentration of less than 150 pg / mL indicates the donor and / or sample is suitable for donation for the production of an Ig preparation.

[0593] Plasma samples or fractions thereof suitable for donation for the production of an Ig preparation will be subjected to one or more further purification steps. Suitable purification steps for the production of an Ig preparation will be apparent to the skilled person and / or described herein. For example, the plasma sample or fraction thereof is subject to one or more steps selected from the group consisting of clarification, ethanol precipitation, octanoic acid fractionation, ammonium sulphate precipitation, affinity chromatography, ion exchange chromatography, viral inactivation, viral filtration, isoagglutinin affinity chromatography, ultrafiltration / diafiltration, bulk formulation, final formulation and combinations thereof.

[0594] In one example, the method of the disclosure is performed before and / or after one or more purification steps selected from a group consisting of: precipitation, such as ethanol, ammonium sulphate and octanoic acid fractionation; affinity chromatography; ion exchange chromatography; viral inactivation; viral filtration; isoagglutinin affinity chromatography, ultrafiltration / diafiltration, bulk formulation, final formulation and combinations thereof.

[0595] In one example, the method of the disclosure is performed before one or more purification steps selected from a group consisting of: precipitation, such as ethanol, ammonium sulphate and octanoic acid fractionation; affinity chromatography; ion exchange chromatography; viral inactivation; viral filtration; isoagglutinin affinity chromatography, ultrafiltration / diafiltration, bulk formulation, final formulation and combinations thereof.

[0596] In one example, the method of the disclosure is performed after one or more purification steps selected from a group consisting of: precipitation, such as ethanol, ammonium sulphate and octanoic acid fractionation; affinity chromatography; ion exchange chromatography; viral inactivation; viral filtration; isoagglutinin affinity chromatography, ultrafiltration / diafiltration, bulk formulation, final formulation and combinations thereof.

[0597] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to a precipitation step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to a precipitation step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to a precipitation step.

[0598] In one example, the precipitation step is ethanol precipitation. For example, cold ethanol may be used to isolate and enrich IgG by removing albumin and a- and P-globulins from the plasma or fractions thereof. For example, as described in WO2011 / 149472.

[0599] In one example, the precipitation step is octanoic acid fractionation. Octanoic acid may be used to remove of plasma lipids and plasma proteins (other than IgG). For example, as described in WO2011 / 131787.

[0600] In one example, the precipitation step is ammonium sulphate precipitation.

[0601] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to an affinity chromatography step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to an affinity chromatography step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to an affinity chromatography step.

[0602] Suitable affinity chromatography steps are also described in W02023 / 007445. For example, an affinity chromatography resin as described herein can be integrated into an affinity purification as described in W02023 / 007445. In one example, an affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig immobilized to a matrix of the affinity chromatography resin, can be integrated into an affinity purification as described in W02023 / 007445.

[0603] In one example, the plasma sample or fraction thereof is subjected to an affinity chromatography resin comprising a ligand capable of specifically binding to a CH3 domain of human IgG. In one example, the plasma sample or fraction thereof is purified using continuous affinity chromatography, the method comprising binding the IgG to an affinity chromatography resin comprising a ligand capable of specifically binding to a CH3 domain of human IgG and collecting the IgG.

[0604] In one example, the ligand comprises a camelid-derived single domain [VHH] antibody fragment. In one example, the resin comprises a matrix selected from the group consisting of a cross-linked poly(styrene-divinylbenzene) matrix and an agarose-based matrix. In one example, the ligand comprises a VHH antibody fragment conjugated to a cross-linked poly(styrene- divinylbenzene) matrix. In one example, the method comprises washing the resin with a wash buffer having a pH of between 5 and 10 and a dissociation constant (pKa) between 6.8 and 8.5 at 25°C. In one example, the method comprises eluting the bound IgG from the resin with an elution buffer having a pH of between 3 and 5. In one example, the wash buffer comprises sodium chloride and / or a divalent salt at a concentration of up to 1000 mM. In one example, the wash buffer comprises 20 mM sodium dihydrogen phosphate, 500 mM sodium chloride and is at a pH of 7.4. In one example, the elution buffer is or comprises a phosphate buffer and / or an acetate buffer at a pH of between 3 and 5. In one example, the equilibration buffer comprises 20 mM sodium dihydrogen phosphate, 500 mM sodium chloride and is at a pH of 7.4. In one example, method for purifying IgG from plasma or a fraction thereof using simulated moving bed (SMB) chromatography, the method comprising: a) equilibrating an affinity chromatography resin comprising a cross-linked poly(styrene- divinylbenzene) matrix and a ligand capable of specifically binding to a CH3 domain of human IgG with a 20mM phosphate equilibration buffer having a pH of between 7 and 8; b) binding the IgG from the plasma or fraction thereof to the resin; c) washing the resin with a 20mM phosphate wash buffer having a pH of between 7 and 8; and d) eluting the bound IgG with a 20mM acetate or phosphate elution buffer having a pH of between 3 and 5; wherein steps a) to d) may be repeated on the affinity chromatography resin and wherein, the affinity chromatography resin is packed into a series of two or more fluidly -connected columns separated by fluid conduits comprising inlet and outlet valves, and optionally wherein the method does not comprise stripping the resin.

[0605] In one example, the method of the disclosure is performed before and / or after purifying IgG from plasma or a fraction thereof using affinity chromatography. In one example, the method of the disclosure is performed before purifying IgG from plasma or a fraction thereof using affinity chromatography. In one example, the method of the disclosure is performed before after purifying IgG from plasma or a fraction thereof using affinity chromatography.

[0606] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to an ion exchange chromatography step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to an ion exchange chromatography step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to an ion exchange chromatography step.

[0607] In one example, the ion exchange chromatography is anion exchange chromatography. For example, anion exchange chromatography may be used to remove IgA, remaining IgM and other plasma components (other than IgG).

[0608] The anion exchanger can be a resin-based anion exchanger, an anion exchange membrane adsorber, or any other format of anion exchanger with a positively charged substrate for capturing negatively charged particles. In one example, the anion exchanger is an anion exchange membrane adsorber. In another example, the anion exchanger is a resin-based anion exchanger. In a further example, the anion exchanger is a monolithic anion exchanger.

[0609] In one example, the sample has been subjected to anion exchange chromatography using a resin-based anion exchanger. For example, the anion exchange chromatography resin is a strong anion exchanger. In one example, the strong anion exchange resin comprises a matrix consisting of a poly(styrene-divinylbenzene) matrix. In one example, the strong anion exchanger comprises a quatemized polyethyleneimine functional group. Suitable resin-based anion exchanges will be apparent to the skilled person and include, for example, POROS™ HQ 50. In one example, the anion exchange chromatography step is performed in flow through mode. In another example, the anion exchange chromatography step is performed in bind-and- elute mode.

[0610] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to a viral inactivation step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to a viral inactivation step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to a viral inactivation step.

[0611] In one example, the viral inactivation may be effected by adjusting the solution to low pH. Low pH may be a pH of between 2 to 4. In one example, low pH viral inactivation is performed in the presence of caprylate. In another example, viral inactivation may be effected by contacting the plasma or fraction thereof, or an IgG-enriched preparation or IgG-containing pharmaceutical composition with w-Octyl-P-D-Glucopyranoside (OG), thereby forming an OG-IgG mixture. In a further example, low pH viral inactivation is performed in the presence of N,N- Dimethylmyristylamine A -ox ide (TDAO).

[0612] In a further example, viral inactivation may be effected by exposing the sample to a solventdetergent inactivation step. Suitable solvent-detergent treatments would be apparent to the skilled person and include, for example environmentally friendly detergents. Exemplary environmentally friendly detergents suitable for use in the present disclosure and in particular for use in inactivating lipid enveloped viruses include AA-Dimethylmyristylamine A -ox ide (TDAO), polysorbate 80 (PS80), polyoxyethylene (10) isooctylcyclohexyl ether (TRITON® X-100-reduced), and a nonionic surfactant prepared from glucose and alcohol (e.g., Simulsol™ formulations). In one example, the detergent is Af,A-Dimethylmyristylamine A -ox ide (TDAO). In one example, the detergent is polysorbate 80. In another example, the detergent is polyoxyethylene (10) isooctylcyclohexyl ether (TRITON® X-100-reduced). In a further example, the detergent is a nonionic surfactant prepared from glucose and alcohol.

[0613] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to a viral filtration step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to a viral filtration step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to a viral filtration step.

[0614] In one example, the viral filtration is performed with membranes of pore sizes from 15-20 nm to remove microbes and viruses from a solution or eluate or pharmaceutical composition. Exemplary nanofilters include Planova S20N (Asahi), Virosart HC (Sartorius) and Planova 20N (Asahi).

[0615] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to an immunoaffinity chromatography step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to an immunoaffinity chromatography step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to an immunoaffinity chromatography step.

[0616] In one example, the immunoaffinity chromatography step is isoagglutinin affinity chromatography. For example, the isoagglutinin affinity chromatography is performed using Eshmuno anti-A and anti-B resin. For example, isoagglutinin affinity chromatography may be used to remove isoagglutinins A and B.

[0617] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to an ultrafiltration / diafiltration step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to an ultrafiltration / diafiltration chromatography step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to an ultrafiltration / diafiltration chromatography step.

[0618] An exemplary ultrafiltration / diafiltration membrane is Pellicon 2 Cassettes (Millipore) or Poly ethersulfone or Hydrosart cassettes (Sartorius).

[0619] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to a bulk formulation step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to a bulk formulation step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to a bulk formulation step.

[0620] In one example, the method of the disclosure is performed before and / or after the sample comprising the target ligand is subjected to a final formulation step. In one example, the method of the disclosure is performed before the sample comprising the target ligand is subjected to a final formulation step. In one example, the method of the disclosure is performed after the sample comprising the target ligand is subjected to a final formulation step. For example, the sample is formulated in the final commercially available formulation.

[0621] In one example, the method of the disclosure is performed on a formulated Ig product. For example, the method of the disclosure is performed on a polyclonal IgG selected from the group consisting of Hizentra® (CSL Behring), Privigen® (CSL Behring), Flebogamma® (Grifols), Gamunex®-C (Grifols), Gammagard® (Takeda), and Octagam® (Octapharma). Gamunex®-C (Grifols), Xembify® (Grifols), Cutaquig® (Octapharma) and Cuvitru® (Takeda).

[0622] Anti-FcsRI Ig reduced plasma preparations and anti-FcsRI Ig reduced IgG preparations of the disclosure are useful for formulations into a pharmaceutical composition for parenteral, such as intravenous administration or subcutaneous administration, for therapeutic and prophylactic treatment.

[0623] A pharmaceutical composition of present invention according to a preferred embodiment comprises the anti-FcsRI Ig reduced preparation or the anti-FcsRI Ig reduced IgG plasma preparation.

[0624] The compositions for administration will commonly comprise a solution of the anti-FcsRI Ig reduced preparation or anti-FcsRI Ig reduced IgG preparation of the disclosure dissolved in a pharmaceutically acceptable carrier, such as an aqueous carrier. A variety of aqueous carriers can be used, e.g., buffered saline and the like. The compositions may contain pharmaceutically acceptable carriers as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents and the like, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like.

[0625] The concentration of the Ig in the pharmaceutical composition of the present disclosure in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight and the like in accordance with the particular mode of administration selected and the patient's needs. The vehicles may contain minor amounts of additives that enhance isotonicity and chemical stability, e.g., buffers and preservatives. For example, the pharmaceutical composition comprises proline as a stabilising agent.

[0626] Suitable pharmaceutical compositions in accordance with the disclosure will generally include an amount of the anti-FcsRI Ig reduced preparation of the present disclosure admixed with an acceptable pharmaceutical carrier, such as a sterile aqueous solution, to give a range of final concentrations, depending on the intended use. The techniques of preparation are generally known in the art as exemplified by Remington's Pharmaceutical Sciences, 16th Ed. Mack Publishing Company, 1980.

[0627] For example, the total Ig concentration of the pharmaceutical composition is 1 to 5% w / v, 5 to 15% w / v, or 8 to 12% w / v. For example, the total Ig concentration of the pharmaceutical composition is 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15% w / v. For intravenous use, 1% w / v (i.e. 10g Ig / L) may be used. For intravenous use, 10% w / v (i.e. 100g Ig / L) may be used.

[0628] For subcutaneous administration, a higher concentration may be used. For example, 15 to 35% w / v, or 20 to 30% w / v. In one example, the total Ig concentration of the pharmaceutical composition is 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26% w / v.

[0629] Anti-FcsRI Ig reduced Ig Preparations

[0630] The present disclosure provides an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation, comprising polyclonal IgG.

[0631] The present disclosure also provides an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced IgG preparation, comprising polyclonal IgG.

[0632] In one example, the polyclonal IgG is present in an amount from 5 to 25% (w / v). In one example, the polyclonal IgG is present in an amount from 6 to 15% (w / v). In one example, the polyclonal IgG is present in an amount from about 8 to 12% (w / v). In one example, the polyclonal IgG is present in an amount of about 10% (w / v).

[0633] The final protein concentration will depend on various factors, such as the administration route, the type of condition to be treated, etc. The skilled person will be able to determine the optimal protein concentration for the intended application. For example, for intravenous infusion, the polyclonal IgG is present in an amount from about 15 to 20% (w / v). In one example, the polyclonal IgG is present in an amount from about 8 to 12% (w / v). In another example, the polyclonal IgG is present in an amount of about 20% (w / v).

[0634] For subcutaneous administration a higher dosage may be chosen, for instance about 15 to 20% (w / v). In one example, the polyclonal IgG is present in an amount of about 20% (w / v).

[0635] The immunoglobulins can be isolated from human or animal blood or produced by other means, for instance by recombinant DNA technology or hybridoma technology. In one example, the immunoglobulin is obtained from blood plasma, typically from a pool of blood plasma from many donors. In order to obtain the immunoglobulins from plasma, the plasma is subjected to alcohol fractionation, which may be combined with other purification techniques like chromatography, adsorption or precipitation as described herein. However, other processes can also be used.

[0636] The pharmaceutical compositions of the disclosure are formulated by methods known in the art. In the case of an IgG solution, the pH of the final preparation is adjusted to a relatively high but acidic pH, namely in the range of about pH 4.2 to 5.4. It has been found that this pH range is particularly useful for improving the storage of characteristics of polyclonal IgG preparations. In one example, the pH is from about 4.6 to 5.0. In an example, the pH is 4.8.

[0637] In other examples, the pharmaceutical composition further comprises a stabilizer. In one example, the stabilizer is more than one amino acid. For example, the amino acids are selected from the group consisting of non-polar and basic amino acids. In some examples, the amino acid is selected from the group consisting of histidine, arginine, lysine, ornithine, isoleucine, valine, methionine, glycine and proline.

[0638] In one example, the stabilizer is proline. For example, the proline is L-proline. The amount of proline in the pharmaceutical composition ranges from about 10 to about 2000 mmol / 1. In some examples, the amount of proline in the pharmaceutical composition ranges from about 50 to about 1000 mmol / 1. In other examples, the amount of proline in the pharmaceutical composition ranges from about 100 to 500 mmol / 1.

[0639] In one example, the amount of L-proline in the pharmaceutical composition ranges from about 200 mmol / L to 300 mmol / L. For example, the amount of L-proline in the preparation ranges from about 225 mmol / L to 275 mmol / L. In one example, the amount of L-proline in the pharmaceutical composition ranges from about 240 mmol / L to 260 mmol / L. For example, the amount of L-proline in the pharmaceutical composition ranges from about 250 mmol / L.

[0640] In further examples, the amount of proline in the pharmaceutical composition is about 250 mmol / 1.

[0641] In one example, the final concentration of proline is between 200 mM to 400 mM. In one example, the final concentration of proline is 250 mM.

[0642] In one example, the proline is L-proline. In another example, the proline is a proline equivalent (e.g., proline analogues).

[0643] In other examples, the stabilizer is present in the pharmaceutical composition at a concentration of more than 200 mM. In some examples, the stabilizer is present in the pharmaceutical composition at a concentration between 200 mM and 400 mM. In another examples, the stabilizer is present in the pharmaceutical composition at a concentration between 200 mM and 300 mM. In further examples, the stabilizer is present in the pharmaceutical composition at a concentration of 250 mM.

[0644] In one example, the pharmaceutical composition comprises a pH of between 4 and 5.5. For example, the pharmaceutical composition comprises a pH of between 4.5 and 5.0. In one example, the pharmaceutical composition comprises a pH of between 4.6 and 5.0. For example, the pharmaceutical composition comprises a pH of 4.6. In one example, the pharmaceutical composition or Ig preparation or anti-FcsRI Ig reduced preparation comprises a pH of 4.7. In another example, the pharmaceutical composition comprises a pH of 4.8. In a further example, the pharmaceutical composition comprises a pH of 4.9. In one example, the pharmaceutical composition comprises a pH of 5.0.

[0645] In one example, the pharmaceutical composition comprises 100 mg / mL of total human plasma protein. In one example, the pharmaceutical composition comprises 20 g / 100 mL of total human plasma protein.

[0646] In one example, the pharmaceutical composition comprises a purity of more than 95% immunoglobulin G (IgG). For example, the pharmaceutical composition comprises a purity of more than 96% immunoglobulin G (IgG). In another example, the pharmaceutical composition comprises a purity of more than 97% immunoglobulin G (IgG). In another example, the pharmaceutical composition comprises a purity of more than 98% immunoglobulin G (IgG). In another example, the pharmaceutical composition comprises a purity of more than 99% immunoglobulin G (IgG).

[0647] In one example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgGl subclass distribution of more than 60%. For example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgGl subclass distribution of more than 65%.

[0648] In one example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgG2 subclass distribution of less than 30%. For example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgG2 subclass distribution of less than 28%.

[0649] In one example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgG3 subclass distribution of less than 5%. For example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgG3 subclass distribution of less than 4%.

[0650] In one example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgG4 subclass distribution of less than 5%. For example, the pharmaceutical composition or the anti-FcsRI Ig reduced preparation comprises an IgG4 subclass distribution of less than 3%. In one example, the pharmaceutical composition or anti-FcsRI Ig reduced preparation comprises an IgG subclass distribution that is similar to that of normal human plasma, for example 69% IgGi, 26% IgG2, 3% IgG3and 2% IgG4.

[0651] In one example, the pharmaceutical composition comprises a nominal osmolality of between about 300 mOsm / kg and 400 mOsm / kg. In one example, the pharmaceutical composition comprises a nominal osmolality of 380 mOsm / kg. For example, the pharmaceutical composition comprises a nominal osmolality of between about 300 mOsm / kg and 350 mOsm / kg. In one example, the pharmaceutical composition comprises a nominal osmolality of 320 mOsm / kg.

[0652] In one example, the pharmaceutical composition comprises a sodium content of < 1 mmol / L.

[0653] In one example, the pharmaceutical composition comprises an IgA content of < 0.05 mg / mL. For example, the pharmaceutical composition comprises an IgA content of < 0.04 mg / mL, or < 0.03 mg / mL. In one example, the pharmaceutical composition comprises an IgA content of < 0.025 mg / mL. In one example, the pharmaceutical composition comprises an IgA content of < 0.01 mg / mL. For example, the pharmaceutical composition comprises an IgA content of < 0.009 mg / mL.

[0654] In one example, the pharmaceutical composition comprises an IgA content of < 0.1 mg / g IgG.

[0655] In one example, the pharmaceutical composition comprises an IgA content of <0.09 mg / g IgG.

[0656] In one example, the pharmaceutical composition comprises an IgM content of < 10 mg / L. For example, an IgM content of < 10 mg / L, < 9 mg / L, < 8 mg / L, < 7 mg / L, < 6 mg / L, < 5 mg / L, < 4 mg / L, < 3 mg / L, < 2 mg / L. In one example, the pharmaceutical composition comprises an IgM content of < 2 mg / L. In one example, the pharmaceutical composition comprises an IgM content of < 1 mg / L. In one example, the pharmaceutical composition comprises an IgM content of < 0.5 mg / L. For example, the pharmaceutical composition comprises an IgM content of <0.17 mg / L. In one example, the pharmaceutical composition comprises an IgM content of < 2 pg / g IgG.

[0657] In one example, the pharmaceutical composition comprises an IgM content of < 1.9 pg / g IgG.

[0658] In one example, the pharmaceutical composition comprises an albumin content of < 0.50 mg / mL. For example, the pharmaceutical composition comprises an albumin content of < 0.40 mg / mL. In one example, the pharmaceutical composition or anti-FcsRI Ig reduced Ig preparation comprises an albumin content of < 0.30 mg / mL. In one example, the pharmaceutical composition comprises an albumin content of < 0.20 mg / mL. In one example, the pharmaceutical composition comprises an albumin content of < 0.10 mg / mL. For example, the pharmaceutical composition comprises an albumin content of < 0.09 mg / mL. In one example, the pharmaceutical composition comprises an albumin content of < 0.08 mg / mL. In one example, the pharmaceutical composition comprises an albumin content of < 0.07 mg / mL. In one example, the pharmaceutical composition comprises an albumin content of < 1 mg / g IgG. In one example, the pharmaceutical composition comprises an albumin content of < 0.80 mg / g IgG.

[0659] In one example, the pharmaceutical composition comprises a Prekallikrein activator (PKA) level of < 35 lU / mL. In one example, the pharmaceutical composition comprises a Prekallikrein activator (PKA) level of < 30 HJ / mL. In one example, the pharmaceutical composition comprises a Prekallikrein activator (PKA) level of < 50 lU / mL. In one example, the pharmaceutical composition comprises a Prekallikrein activator (PKA) level of < 20 lU / mL. For example, the pharmaceutical composition comprises a Prekallikrein activator (PKA) level of < 15 lU / mL. In one example, the pharmaceutical composition comprises a Prekallikrein activator (PKA) level of < 10 lU / mL.

[0660] In one example, the anti-FcsRI Ig reduced preparation comprises polyclonal IgG at an amount of 10% (w / v), proline at a concentration of 250 mM, and a pH of 4.8.

[0661] In one example, the anti-FcsRI Ig reduced preparation comprises polyclonal IgG at an amount of 20% (w / v), proline at a concentration of 250 mM, PS80 at an amount of 20 pg / mL, and a pH of 4.8.

[0662] Exemplary methods of purifying Ig processes are described in W02015 / 000886, exemplary IgG products are described in WO2016 / 087569, both of which are incorporated herein by reference.

[0663] The present disclosure also provides a pooled IgG preparation, wherein the pooled IgG preparation comprises a detectable amount of an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig). It will be apparent to the skilled person from the disclosure herein that the pooled IgG preparation with a detectable amount of an anti-FcsRI Ig has not been subjected to any method of the present disclosure. Accordingly, the present disclosure also provides a method of detecting anti-FcsRI Ig in a pooled IgG preparation.

[0664] In one example, the pooled IgG preparation comprises other isotypes such IgA and IgM. In one example, the pooled IgG preparation comprises IgA. In one example, the pooled IgG preparation comprises other isotypes IgM.

[0665] Method of use

[0666] As discussed herein, the present disclosure provides a method of treating, preventing and / or delaying progression of a condition relating to a primary or secondary immune deficiency, an inflammatory disease, an autoimmune disease or an acute infection in a subject, comprising administering an anti-FcsRI Ig reduced preparation, an anti-FcsRI Ig reduced IgG preparation or a pharmaceutical formulation to the subject.

[0667] The present disclosure also provides an anti-FcsRI Ig reduced preparation or an anti-FcsRI Ig reduced IgG preparation or a pharmaceutical composition described herein for use in treating, preventing and / or delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject. The present disclosure further provides an anti-FcsRI Ig reduced preparation or an anti- FcsRI Ig reduced IgG preparation or a pharmaceutical composition described herein in the manufacture of a medicament for treating, preventing and / or delaying progression of a condition relating to an immune deficiency, an inflammatory disease, an autoimmune disease and / or an acute infection in a subject.

[0668] In one example, the condition is selected from a group consisting of primary immunodeficiency disease (PI), chronic inflammatory demyelinating polyneuropathy (CIDP), chronic immune thrombocytopenic purpura (ITP) and combinations thereof.

[0669] In one example the condition is primary immunodeficiency disease (PI).

[0670] In one example, the condition is chronic inflammatory demyelinating polyneuropathy (CIDP).

[0671] In one example, the condition is chronic immune thrombocytopenic purpura (ITP).

[0672] In another example, the condition is selected from the group consisting of congenital agammaglobulinaemia and hypogammaglobulinaemia, common variable immunodeficiency, severe combined immunodeficiency, allogenic bone marrow transplant, chronic lymphocytic leukaemia, pediatric HIV, kidney transplant with a high antibody recipient or with an ABO incompatible donor, chronic fatigue syndrome, Clostridium difficile colitis, Graves' ophthalmopathy, muscular dystrophy, inclusion body myositis, Lambert-Eaton syndrome, Lupus erythematosus, multifocal motor neuropathy, myasthenia gravis, neonatal alloimmune thrombocytopenia, Parvovirus B19 infection, pemphigus, post-transfusion purpura, renal transplant rejection, spontaneous Abortion Miscarriage, stiff person syndrome, opsoclonus Myoclonus, severe sepsis and septic shock in critically ill adults, toxic epidermal necrolysis, chronic lymphocytic leukemia, multiple myeloma, X-linked agammaglobulinemia, hypogammaglobulinemia, RRMS, IgG subclass deficiencies with recurrent infections and combinations thereof.

[0673] In further examples, the treatment further comprises a replacement therapy in myeloma or chronic lymphocytic leukaemia with severe secondary hypogammaglobulinaemia and recurrent infections.

[0674] In other examples, the condition is selected from a group consisting of autoimmune diseases and certain neurological diseases, such as Rheumatoid arthritis, Systemic Lupus Erythematosus (SLE), Antiphospholipid syndrome, immune thrombocytopenia (ITP), Kawasaki disease, Guillain Barre syndrome (GBS), multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CIDP), multifocal motor neuropathy (MMN), myasthenia gravis (MG), skin blistering diseases, scleroderma, Dermatomyositis, Polymyositis, Alzheimer's Disease, Parkinson's Disease, Alzheimer's Disease related to Downs Syndrome, cerebral amyloid angiopathy, Dementia with Lewy bodies, Fronto-temporal lobar degeneration, vascular dementia, cell and organ transplant and combinations thereof.

[0675] In some examples, the pharmaceutical composition is present in a vial, a prefilled syringe or an autoinjector device. The present disclosure also provides a prefilled syringe comprising the pharmaceutical composition described herein.

[0676] The present disclosure also provides an autoinjector device comprising the pharmaceutical composition described herein.

[0677] In one example, the composition of the disclosure is administered subcutaneously to the subject in need thereof. In another example, the composition of the disclosure is administered intravenously to the subject in need thereof.

[0678] In one example, the composition of the disclosure is self-administered.

[0679] In one example, the composition of the disclosure is self-administered subcutaneously.

[0680] In one example, the composition of the disclosure is provided in a pre-filled syringe.

[0681] In one example, the composition of the disclosure is self-administered subcutaneously, with a pre-filled syringe.

[0682] In one example of any method described herein, the subject is a mammal, for example a primate such as a human.

[0683] Kits

[0684] The present disclosure further provides a kit containing a first and / or second binding moiety of the disclosure for use in a method described herein.

[0685] The present disclosure further provides a kit for detecting a target ligand of interest, the kit comprising a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label.

[0686] The present disclosure further provides a kit for detecting an anti-FcsRI Ig, the kit comprising a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label.

[0687] In one example, the panel or kit as described herein is for ex vivo analysis. In one example, the kit is suitable for use with a whole blood sample, a plasma sample or fraction thereof.

[0688] Reagents can also be provided in the kits. In one example, the kits comprise a device (e.g., a multi-well plate). In addition, the kit includes various diluents and buffers, or other agents for performing the methods described above. Other components of the kit can easily be determined by one of skill in the art. Such components may include coating reagents, indicator charts for colorimetric comparisons, disposable gloves, decontamination instructions, applicator sticks or containers, a sample preparatory cup, etc. In one example, a kit comprises buffers or other reagents appropriate for constituting a reaction medium in which the composition disclosed herein is contacted with the sample.

[0689] In one example, the kit further comprise an instruction or package insert with instructions for detecting and / or screening a target ligand in a sample. For example, in certain examples, the kit comprises an instruction indicating how to use the kit detect an anti-FcsRI Ig, or to screen and / or identify plasma samples suitable for administration to a subject, or identify subjects suitable and / or unsuitable for plasma donation for the production of an Ig preparation. In one example, the kit comprises an instruction indicating how to prepare a sample. In one example, the kit provides instructions for contacting the sample with a binding moiety disclosed herein (i.e., FcsRI or fragment or epitope thereof) prior to analysing the sample for the presence of an anti-FcsRI Ig. The kit may also provide instructions for optimization of buffers, optimization of the ratios of the various components, optimization of dilution of the sample, and optimization of the order of the mixture and application steps (e.g., mix all components prior to application, mix only certain components and apply others separately).

[0690] The invention is further disclosed in the following numbered paragraphs:

[0691] 1. A method of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand’s receptor, the method comprising:

[0692] (i) contacting the sample comprising the target ligand to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the target ligand in the sample to thereby form an immunocomplex;

[0693] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0694] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0695] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the target ligand in the sample.

[0696] 2. The method of paragraph 1, wherein prior to contacting the immunocomplex to the first capture probe immobilized on the first solid matrix, the method comprises:

[0697] (i) contacting the immunocomplex to a second capture probe immobilized on a second solid matrix, wherein the second capture probe binds the second nucleic acid tag;

[0698] (ii) washing the second solid matrix to remove unbound immunocomplex; and

[0699] (iii)releasing the bound immunocomplex from the second immobilized capture probe.

[0700] 3. The method of paragraphs 1 and 2, wherein the nucleic acid reporter is generated by:

[0701] (i) linking the first nucleic acid tag and the second nucleic acid tag, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid tag; or

[0702] (ii) linking the first nucleic acid tag and the second nucleic acid label, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid label; or

[0703] (iii) linking the first nucleic acid label and the second nucleic acid tag, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid tag; or

[0704] (iv) linking the first nucleic acid label and the second nucleic acid label, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid label.

[0705] 4. The method of paragraph 3, wherein the nucleic acid reporter is formed by proximity ligation or proximity extension.

[0706] 5. The method of any one of paragraphs 1 to 4, wherein the method further comprises washing the first solid matrix to remove unbound immunocomplex.

[0707] 6. The method of any one of paragraphs 1 to 5, wherein the first capture probe is: (i) a protein that specifically binds to the first nucleic acid tag;

[0708] (ii) a protein and nucleic acid complex that specifically binds to the first nucleic acid tag;

[0709] (iii)a nucleic acid molecule, wherein the first capture probe or a fragment thereof is complementary to the first nucleic acid tag or a fragment thereof; or

[0710] (iv)a nucleic acid molecule, wherein the first capture probe or a fragment thereof hybridizes with the first nucleic acid tag or a fragment thereof.

[0711] 7. The method of any one of paragraphs 1 to 6, wherein the first capture probe comprises a biotin binding protein.

[0712] 8. The method of paragraph 7, wherein the biotinylated tail sequence of the first nucleic acid tag binds to the biotin binding protein.

[0713] 9. The method of paragraph 8, wherein the biotin binding protein is streptavidin, avidin or neutravidin.

[0714] 10. The method of paragraph 9, wherein the biotinylated tail sequence of the first nucleic acid tag binds the streptavidin or avidin of the first capture probe.

[0715] 11. The method of any one of paragraphs 1 to 10, wherein the first nucleic acid tag binds to the first capture probe via a thioester group, a disulfide linkage, a cleavable linkage or a proteinprotein interaction.

[0716] 12. The method of any one of paragraphs 2 to 11, wherein the second capture probe is:

[0717] (i) a protein that specifically binds to the second nucleic acid tag;

[0718] (ii) a protein and nucleic acid complex that specifically binds to the second nucleic acid tag;

[0719] (iii)a nucleic acid molecule, wherein the second capture probe or a fragment thereof is complementary to the second nucleic acid tag or a fragment thereof; or

[0720] (iv)a nucleic acid molecule, wherein the second capture probe or a fragment thereof hybridizes with the second nucleic acid tag or a fragment thereof.

[0721] 13. The method of any one of paragraphs 2 to 12, wherein the second capture probe comprises a T, an A, a C and / or a G rich sequence complementary to the A, a T, a G and / or a C rich tail sequence of the second nucleic acid tag.

[0722] 14. The method of claim 13, wherein the A, T, G and / or C rich tail sequence of the second nucleic acid tag binds the complementary T, A, C and / or G rich sequence. 15. The method of any one of paragraphs 2 to 14, wherein the second nucleic acid tag binds to the second capture probe via a thioester group, a disulfide linkage, a cleavable linkage or a proteinprotein interaction.

[0723] 16. The method of any one of paragraphs 11 to 15, wherein the cleavable linkage is a photocleavable linkage, a chemically cleavable linkage, or an enzymatically cleavable linkage.

[0724] 17. The method of any one of paragraphs 1 to 16, wherein:

[0725] (i) the first nucleic acid tag is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. fused to the first receptor, fragment or epitope thereof; c. conjugated to the first receptor, fragment or epitope thereof; d. non-covalently bound to the first receptor, fragment or epitope thereof; e. conjugated to the first nucleic acid label; f. non-covalently bound to the first nucleic acid label; or g. part of the first nucleic acid label; and / or

[0726] (ii) the second nucleic acid tag is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. fused to the second receptor, fragment or epitope thereof; c. conjugated to the second receptor, fragment or epitope thereof; d. non-covalently bound to the second receptor, fragment or epitope thereof; e. conjugated to the second nucleic acid label; f. non-covalently bound to the second nucleic acid label; or g. part of the second nucleic acid label.

[0727] 18. The method of any one of paragraphs 1 to 17, wherein:

[0728] (i) the first nucleic acid label is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. conjugated to the first receptor, fragment or epitope thereof; c. non-covalently bound to the first receptor, fragment or epitope thereof; d. conjugated to the first nucleic acid tag; e. non-covalently bound to the first nucleic acid tag; f. hybridizes with the first nucleic acid tag; or g. part of the first nucleic acid tag; and / or

[0729] (ii) the second nucleic acid label is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. conjugated to the second receptor, fragment or epitope thereof; c. non-covalently bound to the second receptor, fragment or epitope thereof; d. conjugated to the second nucleic acid tag; e. non-covalently bound to the second nucleic acid tag; f. hybridizes with the second nucleic acid tag; or g. part of the second nucleic acid tag.

[0730] 19. The method of any one of paragraphs 1 to 18, wherein:

[0731] (i) the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment or epitope thereof; and / or

[0732] (ii) the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment or epitope thereof.

[0733] 20. The method of paragraph 19, wherein the nucleic acid reporter comprises the first receptor barcode and / or second receptor barcode.

[0734] 21. The method of paragraph 20, wherein:

[0735] (i) the first receptor barcode in the nucleic acid reporter is a complementary sequence of the first receptor barcode in the first nucleic acid label; and / or

[0736] (ii) the second receptor barcode in the nucleic acid reporter is a complementary sequence of the second receptor barcode in the second nucleic acid label.

[0737] 22. The method of any one of paragraphs 19 to 21, wherein the first receptor barcode and the second receptor barcode are identical.

[0738] 23. The method of any one of paragraphs 1 to 22, wherein the first nucleic acid label and / or the second nucleic acid label further comprise a sample barcode.

[0739] 24. The method of paragraph 23, wherein the nucleic acid reporter comprises the sample barcode.

[0740] 25. The method of paragraphs 23 or 24, wherein the nucleic acid reporter comprises the first receptor barcode, the second receptor barcode and the sample barcode.

[0741] 26. The method of any one of paragraphs 1 to 25, wherein the method comprises detecting the nucleic acid reporter by polymerase chain reaction (PCR), next generation sequencing, Rolling Cycle Amplification (RCA), strand displacement amplification (SDA), Loop-Mediated Isothermal Amplification (LAMP), Recombinase Polymerase Amplification (RPA), or a QuantiGene assay.

[0742] 27. The method of paragraph 26, wherein the PCR is multiplexed quantitative PCR (qPCR) or multiplexed digital PCR.

[0743] 28. The method of any one of paragraphs 1 to 27, wherein the first capture probe is directly coupled to the first solid matrix. 29. The method of any one of paragraphs 1 to 28, wherein the first solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multi-well plate, an affinity column, a membrane or combinations thereof.

[0744] 30. The method of any one of paragraphs 2 to 29, wherein the second capture probe is directly coupled to the second solid matrix.

[0745] 31. The method of any one of paragraphs 2 to 30, wherein the bound immunocomplex is released from the second immobilized capture probe by increasing the temperature to 70°C.

[0746] 32. The method of any one of paragraphs 2 to 31, wherein the second solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multiwell plate, an affinity column, a membrane or combinations thereof.

[0747] 33. The method of paragraphs 29 or 32, wherein the membrane is a PDVF membrane or a nitrocellulose membrane.

[0748] 34. The method of any one of paragraphs 1 to 33, wherein the method comprises forming the immunocomplex in a solution prior to capturing the immunocomplex on the first solid matrix.

[0749] 35. The method of any one of paragraphs 1 to 34, wherein the immunocomplex is formed in a solution and captured on the first solid matrix simultaneously.

[0750] 36. The method of any one of paragraphs 1 to 35, wherein the first and second binding domains of the target ligand are identical sequences, are different sequences and / or have at least 75% sequence identity.

[0751] 37. The method of any one of paragraphs 1 to 36, wherein the first receptor, fragment or epitope thereof and the second receptor, fragment or epitope thereof are identical receptors, fragments or epitopes thereof.

[0752] 38. The method of any one of paragraphs 1 to 37, wherein the first and / or second receptor, fragment or epitope thereof is selected from the group consisting of a Fc receptor, fragment or epitope thereof; a T cell receptor, fragment or epitope thereof; a B cell receptor, fragment or epitope thereof; an epidermal growth factor (EGF) receptor, fragment or epitope thereof; a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof; an insulin receptor, fragment or epitope thereof; a programmed death 1 (PD-1) receptor, fragment or epitope thereof; and combinations thereof. 39. The method of paragraph 38, wherein:

[0753] (i) the Fc receptor, fragment or epitope thereof is a Fc-gamma receptor (FcyR), a Fc-alpha receptor (FcaR), a Fc-epsilon receptor (FcaR) and / or a polymeric immunoglobulin receptor (plgR); and / or

[0754] (ii) the TNFR superfamily, fragment or epitope thereof is a TNFR1, a TNFR2, lymphotoxin beta receptor, 0X40, cluster of differentiation (CD) 40 (CD40), Fas receptor, a decoy receptor, CD27, CD30, 4-1BB, a death receptor, receptor activator of nuclear factor K B (RANK), osteoprotegerin, TNF-related weak inducer of apoptosis (TWEAK) receptor, transmembrane activator and CAML interactor (TACI), B-cell activating factor (BAFF) receptor and / or herpesvirus entry mediator.

[0755] 40. The method of paragraph 39, wherein:

[0756] (i) the FcyR is a FcyRI, a FcyRIIA, a FcyRIIB, a FcyRIIIA, or a FcyRIIIB;

[0757] (ii) the FcaR is FcaRI; and / or

[0758] (iii)the FcaR is FcaRI or FcaRII.

[0759] 41. The method of paragraph 40, wherein the FcaRI is an a-chain of FcaRI or subunit thereof, or a P-chain of FcaRI or subunit thereof.

[0760] 42. The method of paragraph 41, wherein:

[0761] (i) the first receptor, fragment or epitope thereof is an a-chain of FcaRI or subunit thereof and the second receptor, fragment or epitope thereof is an a-chain of FcaRI or subunit thereof;

[0762] (ii) the first receptor, fragment or epitope thereof is an a-chain of FcaRI or subunit thereof and the second receptor, fragment or epitope thereof is an P-chain of FcaRI or subunit thereof;

[0763] (iii)the first receptor, fragment or epitope thereof is an P-chain of FcaRI or subunit thereof and the second receptor, fragment or epitope thereof is an P-chain of FcaRI or subunit thereof; or

[0764] (iv)the first receptor, fragment or epitope thereof is an P-chain of FcaRI or subunit thereof and the second receptor, fragment or epitope thereof is an a-chain of FcaRI or subunit thereof.

[0765] 43. The method of any one of paragraphs 1 to 38, wherein the target ligand is an immunoglobulin (Ig), a peptide-MHC complex and / or an antigen.

[0766] 44. The method of paragraph 43, wherein the immunoglobulin is an anti-Fc receptor Ig.

[0767] 45. The method of paragraph 44, wherein the anti-Fc receptor Ig is an anti-FcyR Ig, an anti- FcaR Ig or an anti -FcaR Ig.

[0768] 46. The method of paragraph 45, wherein: (i) the anti-FcyR Ig is an anti-FcyRI Ig, an anti-FcyRIIA Ig, an anti-FcyRIIB Ig, an anti- FcyRIIIA Ig, or an anti-FcyRIIIB Ig;

[0769] (ii) the anti-FcaR Ig is an anti-FcaRI Ig; and / or

[0770] (iii)the anti-FcaR Ig is an anti-FcaRI Ig or an anti-FcsRII Ig.

[0771] 47. The method of paragraph 43, wherein the antigen is selected from the group consisting of an EGF, a programmed death ligand 1 (PD-L1), aPD-L2, a TNF-a, a TNF-C, an OX40L, a CD 154, a FasL, a CD70, a Siva, a CD153, a 4-1BB ligand, a TNF-related apoptosis inducing ligand (TRAIL), a RANK ligand (RANKL), TWEAK, a proliferation inducing ligand (APRIL), BAFF, a calcium modulating ligand (CAMLG), LIGHT, TNF-like ligand 1A (TL1A) and combinations thereof.

[0772] 48. A method of detecting an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) in a sample, the method comprising:

[0773] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcaRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcaRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcaRI Ig in the sample to thereby form an immunocomplex;

[0774] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0775] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0776] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0777] 49. A method of screening a sample for an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig), the method comprising:

[0778] (i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;

[0779] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0780] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0781] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample.

[0782] 50. A method of identifying a sample suitable for administration to a subject, the method comprising:

[0783] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, A T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0784] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag; (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0785] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0786] 51. A method of screening a sample to determine suitability of the sample for administration to a subject, the method comprising:

[0787] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0788] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0789] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0790] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample indicates the sample is not suitable for administration to the subject.

[0791] 52. The method of any one of paragraphs 48 to 51, wherein the method further comprises performing the method of any one of paragraphs 2 to 37, 41 or 42.

[0792] 53. The method of any one of paragraphs 1 to 52, wherein the sample is serum, plasma, a plasma fraction, a purified or partially purified plasma or plasma fraction thereof, a fermentation broth or a purified or partially purified fermentation broth thereof, a cell culture harvest or a purified or partially purified cell culture harvest thereof or a protein suspension. 54. The method of paragraph 53, wherein the plasma or plasma fraction is selected the group consisting of a human blood plasma sample, an IgG intermediate product, an intravenous immunoglobulin G (IVIG) product, a subcutaneous immunoglobulin G (SCIG) product, a cryorich plasma, a cryo-poor plasma, a Supernatant I (SN I), a Cohn Fraction II (Fr II), a Cohn Fraction II+III (Fr II+III), a Cohn Fraction (I+)II+III ((FrI+)II+III), a Kistler / Nitschmann Precipitate A (KN A), a Kistler / Nitschmann Precipitate B (KN B), a Kistler / Nitschmann Precipitate of Supernatant B (KN B+l), an IgG depleted intermediate product, a purified IgG product, a purified formulated IgG product, a formulated plasma fractionation product and combinations thereof.

[0793] 55. The method of paragraphs 53 or 54, wherein the purified or partially purified plasma or plasma fraction thereof has been subjected to one or more steps selected from the group consisting of clarification, ethanol precipitation, octanoic acid fractionation, ammonium sulphate precipitation, affinity chromatography, ion exchange chromatography, viral inactivation, viral filtration, isoagglutinin affinity chromatography, ultrafiltration / diafiltration, bulk formulation, final formulation and combinations thereof.

[0794] 56. The method of any one of paragraphs 46 to 55, wherein the method further comprises administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 500 pg / mL, less than 450 pg / mL less than 400 pg / mL, less than 350 pg / mL, less than 300 pg / mL, less than 250 pg / mL, less than 200 pg / mL or less than 150 pg / mL.

[0795] 57. The method of any one of paragraphs 46 to 56, wherein the method further comprises including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti- FcsRI Ig is present in the sample at a concentration of less than 500 pg / mL, less than 450 pg / mL less than 400 pg / mL, less than 350 pg / mL, less than 300 pg / mL, less than 250 pg / mL, less than 200 pg / mL or less than 150 pg / mL.

[0796] 58. The method of any one of paragraphs 46 to 56, wherein the method further comprises excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti- FcsRI Ig is present in the sample at a concentration of greater than 150 pg / mL, greater than 200 pg / mL, greater than 250 pg / mL, greater than 300 pg / mL, greater than 350 pg / mL, greater than 400 pg / mL, greater than 450 pg / mL or greater than 500 pg / mL.

[0797] 59. A method of identifying a sample suitable for use in producing an immunoglobulin (Ig) preparation for administration to a subject, the method comprising:

[0798] (i) contacting the sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0799] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0800] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0801] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 500 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject.

[0802] 60. A method of screening a subj ect to determine suitability for plasma donation for production of an immunoglobulin (Ig) preparation, the method comprising:

[0803] (i) contacting a sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0804] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0805] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 500 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0806] 61. A method of identifying a subject suitable for plasma donation for production of an immunoglobulin (Ig) preparation, the method comprising:

[0807] (i) contacting a sample with a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI, fragment or epitope thereof that specifically binds to a first binding domain of the anti-FcsRI Ig; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI, fragment or epitope thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to an anti-Fc-epsilon receptor I (FcsRI) immunoglobulin (Ig) if present in the sample to thereby form an immunocomplex;

[0808] (ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;

[0809] (iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and

[0810] (iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample; and wherein the presence of the anti-FcsRI Ig in the sample at a concentration of greater than 500 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0811] 62. The method of any one of paragraphs 48 to 61, wherein the anti-FcsRI Ig in the sample activates a FcsRI mediated signaling pathway if administered to a subject.

[0812] 63. The method of any one of paragraphs 48 to 62, wherein anti-FcsRI Ig at a concentration of greater than 500 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject.

[0813] 64. The method of paragraphs 62 or 63, wherein activation of the FcsRI mediated signaling pathway induces basophil activation. 65. The method of any one of paragraphs 62 to 64, wherein activation of the FcsRI mediated signaling pathway induces mast cell activation.

[0814] 66. The method of any one of paragraphs 62 to 65, wherein activation of the FcsRI mediated signaling pathway induces basophil degranulation.

[0815] 67. The method of any one of paragraphs 62 to 66, wherein activation of the FcsRI mediated signaling pathway induces mast cell degranulation.

[0816] 68. The method of any one of paragraphs 48 to 67, wherein the anti-FcsRI Ig is an anti-FcsRI IgG, an anti-FcsRI IgE, an anti-FcsRI IgM and / or an anti-FcsRI IgA.

[0817] 69. The method of paragraph 68, wherein the anti-FcsRI IgG is an IgGl, an IgG2, an IgG3 or an IgG4.

[0818] 70. The method of any one of paragraphs 1 to 69, wherein the method further comprises diluting the sample in an assay solution prior to contacting with the first binding moiety and the second binding moiety.

[0819] 71. The method of paragraph 70, wherein the sample is diluted in the assay solution at a ratio of between 1 : 10 to 1 :20.

[0820] 72. The method of paragraph 70 or 71, wherein the sample is in the assay solution at a concentration of 10% (v / v).

[0821] 73. The method of any one of paragraphs 59 and 62 to 72, wherein the presence of the anti- FcsRI Ig in the sample at a concentration of greater than 150 pg / mL, greater than 200 pg / mL, greater than 250 pg / mL, greater than 300 pg / mL, greater than 350 pg / mL, greater than 400 pg / mL or greater than 450 pg / mL indicates the sample is not suitable for use in producing an Ig preparation for administration to the subject.

[0822] 74. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 150 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0823] 75. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 200 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation. 76. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 250 pg / m indicates the donor is not suitable for plasma donation for the production of an Ig preparation L.

[0824] 77. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 300 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0825] 78. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 350 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0826] 79. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 400 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0827] 80. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 450 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0828] 81. The method of any one of paragraphs 60 to 72, wherein the presence of the anti-FcsRI Ig in the sample at a concentration of or greater than 500 pg / mL indicates the donor is not suitable for plasma donation for the production of an Ig preparation.

[0829] 82. The method of any one of paragraphs 1 to 81, wherein the method further comprises preparing an anti-FcsRI Ig reduced preparation from the sample if anti-FcsRI Ig is present in the sample, the method further comprising binding the anti-FcsRI Ig to a chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the sample and collecting the anti-FcsRI Ig reduced preparation.

[0830] 83. The method of any one of paragraphs 1 to 83, wherein the method further comprises preparing an anti-FcsRI Ig reduced preparation from the sample if anti-FcsRI Ig is present in the sample, the method further comprising:

[0831] (i) loading the sample onto a chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the sample; and

[0832] (ii) collecting the anti-FcsRI Ig reduced preparation.

[0833] 84. An affinity chromatography resin comprising a ligand which binds to anti-FcsRI Ig immobilized to a matrix of the affinity chromatography resin. 85. The affinity chromatography resin of paragraph 84, wherein the resin further comprises a blood group A antigen and a blood group B antigen immobilized to the matrix of the affinity chromatography resin.

[0834] 86. A method of preparing an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation from a plasma sample or a fraction thereof, the method comprising binding the anti- FcsRI Ig to an affinity chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the plasma sample or fraction thereof and collecting the anti-FcsRI Ig reduced preparation.

[0835] 87. The method of paragraph 83 or 86, wherein the method further comprises loading the plasma sample or fraction thereof onto the affinity chromatography resin.

[0836] 88. A method of preparing an anti-Fc epsilon Receptor I (FcsRI) immunoglobulin (Ig) reduced preparation from a plasma sample or a fraction thereof, the method comprising:

[0837] (i) loading the plasma sample or fraction thereof onto an affinity chromatography resin comprising a ligand which binds to the anti-FcsRI Ig in the plasma sample or fraction thereof immobilized to a matrix of the affinity chromatography resin; and

[0838] (ii) collecting the anti-FcsRI Ig reduced preparation.

[0839] 89. The method of any one of paragraphs 83 and 86 to 88, wherein the method further comprises a wash and / or elution step.

[0840] 90. The method of any one of paragraphs 83 and 86 to 89, wherein the method further comprises an elution step.

[0841] 91. The method of any one of paragraphs 83 and 86 to 90, wherein the method further comprises a wash step.

[0842] 92. The method of any one of paragraphs 83 and 86 to 91, wherein the anti-FcsRI Ig binds to an a-chain of the FcsRI and activates a FcsRI mediated signalling pathway.

[0843] 93. The method of any one of paragraphs 83 and 86 to 92, wherein the anti-FcsRI Ig reduced preparation induces reduced activation of a FcsRI mediated signalling pathway relative to a preparation wherein the anti-FcsRI Ig is not reduced.

[0844] 94. The method of any one of paragraphs 83 and 86 to 93, wherein the anti-FcsRI Ig is not present in a detectable amount in the anti-FcsRI Ig reduced preparation.

[0845] 95. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 500 pg / mL of anti-FcsRI Ig. 96. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 400 pg / mL of anti-FcsRI Ig.

[0846] 97. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 350 pg / mL of anti-FcsRI Ig.

[0847] 98. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 300 pg / mL of anti-FcsRI Ig.

[0848] 99. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 250 pg / mL of anti-FcsRI Ig.

[0849] 100. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 200 pg / mL of anti-FcsRI Ig.

[0850] 101. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig reduced preparation comprises less than 150 pg / mL of anti-FcsRI Ig.

[0851] 102. The method of paragraph 94, wherein the anti-FcsRI Ig reduced preparation induces reduced activation of a FcsRI mediated signalling pathway relative to a preparation wherein the anti-FcsRI Ig is not reduced.

[0852] 103. The method of any one of paragraphs 83 and 86 to 94, wherein the anti-FcsRI Ig is not present in a detectable amount and the anti-FcsRI Ig reduced preparation induces reduced basophil activation relative to a preparation wherein the anti-FcsRI Ig is not reduced.

[0853] 104. The method of any one of paragraphs 83 and 86 to 94, whe...

Claims

CLAIMS1. A method of detecting a target ligand in a sample, wherein the target ligand is capable of inducing oligomerization and / or cross-linking of the target ligand’s receptor, the method comprising:(i) contacting the sample comprising the target ligand to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first receptor, fragment or epitope thereof that specifically binds to a first binding domain of the target ligand; ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second receptor, fragment or epitope thereof that specifically binds to a second binding domain of the target ligand, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the target ligand in the sample to thereby form an immunocomplex;(ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;(iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and(iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the target ligand in the sample, optionally wherein prior to contacting the immunocomplex to the first capture probe immobilized on the first solid matrix, the method comprises:(i) contacting the immunocomplex to a second capture probe immobilized on a second solid matrix, wherein the second capture probe binds the second nucleic acid tag;(ii) washing the second solid matrix to remove unbound immunocomplex; and(iii)releasing the bound immunocomplex from the second immobilized capture probe.

2. The method of claim 1, wherein the nucleic acid reporter is generated by:(i) linking the first nucleic acid tag and the second nucleic acid tag, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid tag; or(ii) linking the first nucleic acid tag and the second nucleic acid label, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid tag and a fragment of the second nucleic acid label; or(iii) linking the first nucleic acid label and the second nucleic acid tag, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid tag; or(iv) linking the first nucleic acid label and the second nucleic acid label, and detecting the nucleic acid reporter composed of a fragment of the first nucleic acid label and a fragment of the second nucleic acid label, optionally wherein the nucleic acid reporter is formed by proximity ligation or proximity extension.

3. The method of claim 1, wherein:(A) the first capture probe is:(i) a protein that specifically binds to the first nucleic acid tag;(ii) a protein and nucleic acid complex that specifically binds to the first nucleic acid tag;(iii)a nucleic acid molecule, wherein the first capture probe or a fragment thereof is complementary to the first nucleic acid tag or a fragment thereof; or(iv)a nucleic acid molecule, wherein the first capture probe or a fragment thereof hybridizes with the first nucleic acid tag or a fragment thereof; and / or(B) the second capture probe is:(i) a protein that specifically binds to the second nucleic acid tag;(ii) a protein and nucleic acid complex that specifically binds to the second nucleic acid tag;(iii)a nucleic acid molecule, wherein the second capture probe or a fragment thereof is complementary to the second nucleic acid tag or a fragment thereof; or(iv)a nucleic acid molecule, wherein the second capture probe or a fragment thereof hybridizes with the second nucleic acid tag or a fragment thereof.

4. The method of claim 1, wherein:(i) the first capture probe comprises a biotin binding protein, optionally wherein the biotinylated tail sequence of the first nucleic acid tag binds to the biotin binding protein; and / or(ii) the second capture probe comprises a T, an A, a C and / or a G rich sequence complementary to the A, a T, a G and / or a C rich tail sequence of the second nucleic acid tag, optionally wherein the A, T, G and / or C rich tail sequence of the second nucleic acid tag binds the complementary T, A, C and / or G rich sequence.

5. The method of claim 4, wherein the biotin binding protein is streptavidin, avidin or neutravidin, optionally wherein the biotinylated tail sequence of the first nucleic acid tag binds the streptavidin or avidin of the first capture probe.

6. The method of claim 1, wherein:(i) the first nucleic acid tag binds to the first capture probe via a thioester group, a disulfide linkage, a cleavable linkage or a protein-protein interaction; and / or(ii) the second nucleic acid tag binds to the second capture probe via a thioester group, a disulfide linkage, a cleavable linkage or a protein-protein interaction, optionally wherein the cleavable linkage is a photocleavable linkage, a chemically cleavable linkage, or an enzymatically cleavable linkage.

7. The method of claim 1, wherein:(i) the first nucleic acid tag is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. fused to the first receptor, fragment or epitope thereof; c. conjugated to the first receptor, fragment or epitope thereof; d. non-covalently bound to the first receptor, fragment or epitope thereof; e. conjugated to the first nucleic acid label; f. non-covalently bound to the first nucleic acid label; or g. part of the first nucleic acid label; and / or(ii) the second nucleic acid tag is: a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. fused to the second receptor, fragment or epitope thereof; c. conjugated to the second receptor, fragment or epitope thereof; d. non-covalently bound to the second receptor, fragment or epitope thereof; e. conjugated to the second nucleic acid label; f. non-covalently bound to the second nucleic acid label; or g. part of the second nucleic acid label; and / or(iii)the first nucleic acid label is: a. directly or indirectly bound to the first receptor, fragment or epitope thereof; b. conjugated to the first receptor, fragment or epitope thereof; c. non-covalently bound to the first receptor, fragment or epitope thereof; d. conjugated to the first nucleic acid tag; e. non-covalently bound to the first nucleic acid tag; f. hybridizes with the first nucleic acid tag; or g. part of the first nucleic acid tag; and / or(iv)the second nucleic acid label is:a. directly or indirectly bound to the second receptor, fragment or epitope thereof; b. conjugated to the second receptor, fragment or epitope thereof; c. non-covalently bound to the second receptor, fragment or epitope thereof; d. conjugated to the second nucleic acid tag; e. non-covalently bound to the second nucleic acid tag; f. hybridizes with the second nucleic acid tag; or g. part of the second nucleic acid tag.

8. The method of claim 1, wherein:(i) the first nucleic acid label further comprises a first receptor barcode specific to the first receptor, fragment or epitope thereof; and / or(ii) the second nucleic acid label comprises a second receptor barcode specific to the second receptor, fragment or epitope thereof, optionally wherein the nucleic acid reporter comprises the first receptor barcode and / or second receptor barcode and wherein:(i) the first receptor barcode in the nucleic acid reporter is a complementary sequence of the first receptor barcode in the first nucleic acid label; and / or(ii) the second receptor barcode in the nucleic acid reporter is a complementary sequence of the second receptor barcode in the second nucleic acid label, optionally wherein the first receptor barcode and the second receptor barcode are identical.

9. The method of claim 1, wherein the first nucleic acid label and / or the second nucleic acid label further comprise a sample barcode, optionally wherein the nucleic acid reporter comprises the sample barcode and / or the nucleic acid reporter comprises the first receptor barcode, the second receptor barcode and the sample barcode.

10. The method of claim 1, wherein the method comprises detecting the nucleic acid reporter by polymerase chain reaction (PCR), next generation sequencing, Rolling Cycle Amplification (RCA), strand displacement amplification (SDA), Loop-Mediated Isothermal Amplification (LAMP), Recombinase Polymerase Amplification (RPA), or a QuantiGene assay, optionally wherein the PCR is multiplexed quantitative PCR (qPCR) or multiplexed digital PCR.

11. The method of claim 1, wherein:(i) the first capture probe is directly coupled to the first solid matrix, optionally wherein the first solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multi-well plate, an affinity column, a membrane or combinations thereof; and / or(ii) the second capture probe is directly coupled to the second solid matrix, optionally wherein the second solid matrix is selected from the group consisting of a magnetic and / or paramagnetic bead or particle, a well of a multi-well plate, an affinity column, a membrane or combinations thereof.

12. The method of claim 1, wherein the bound immunocomplex is released from the second immobilized capture probe by increasing the temperature to 70°C.

13. The method of claim 1, wherein the method comprises forming the immunocomplex in a solution prior to capturing the immunocomplex on the first solid matrix.

14. The method of claim 1, wherein the immunocomplex is formed in a solution and captured on the first solid matrix simultaneously.

15. The method of claim 1, wherein:(i) the first and second binding domains of the target ligand are identical sequences, are different sequences and / or have at least 75% sequence identity; and / or(ii) the first receptor, fragment or epitope thereof and the second receptor, fragment or epitope thereof are identical receptors, fragments or epitopes thereof; and / or(iii) the first and / or second receptor, fragment or epitope thereof is selected from the group consisting of a Fc receptor, fragment or epitope thereof; a T cell receptor, fragment or epitope thereof; a B cell receptor, fragment or epitope thereof; an epidermal growth factor (EGF) receptor, fragment or epitope thereof; a tumour necrosis factor (TNF) receptor superfamily, fragment or epitope thereof; an insulin receptor, fragment or epitope thereof; a programmed death 1 (PD-1) receptor, fragment or epitope thereof; and combinations thereof.

16. The method of claim 15, wherein:(i) the Fc receptor, fragment or epitope thereof is a Fc-gamma receptor (FcyR), a Fc-alpha receptor (FcaR), a Fc-epsilon receptor (FcaR) and / or a polymeric immunoglobulin receptor (plgR); optionally wherein: a. the FcyR is a FcyRI, a FcyRIIA, a FcyRIIB, a FcyRIIIA, or a FcyRIIIB; b. the FcaR is FcaRI; and / or c. the FcaR is FcaRI or FcaRII; and / or(ii) the TNFR superfamily, fragment or epitope thereof is a TNFR1, a TNFR2, lymphotoxin beta receptor, 0X40, cluster of differentiation (CD) 40 (CD40), Fas receptor, a decoy receptor, CD27, CD30, 4-1BB, a death receptor, receptor activator of nuclear factor K B (RANK), osteoprotegerin, TNF-related weak inducer of apoptosis (TWEAK) receptor,transmembrane activator and CAML interactor (TACI), B-cell activating factor (BAFF) receptor and / or herpesvirus entry mediator.

17. The method of claim 16, wherein the FcsRI is an a-chain of FcsRI or subunit thereof, or a P-chain of FcsRI or subunit thereof, optionally wherein:(i) the first receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof;(ii) the first receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof;(iii)the first receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof; or(iv)the first receptor, fragment or epitope thereof is an P-chain of FcsRI or subunit thereof and the second receptor, fragment or epitope thereof is an a-chain of FcsRI or subunit thereof.

18. The method of claim 1, wherein the target ligand is an immunoglobulin (Ig), a peptide- MHC complex and / or an antigen, optionally wherein:(i) the immunoglobulin is an anti-Fc receptor Ig; and / or(ii) the antigen is selected from the group consisting of an EGF, a programmed death ligand 1 (PD-L1), a PD-L2, a TNF-a, a TNF-C, an OX40L, a CD 154, a FasL, a CD70, a Siva, a CD153, a 4-1BB ligand, a TNF-related apoptosis inducing ligand (TRAIL), a RANK ligand (RANKL), TWEAK, a proliferation inducing ligand (APRIL), BAFF, a calcium modulating ligand (CAMLG), LIGHT, TNF-like ligand 1A (TL1A) and combinations thereof.

19. The method of claim 18, wherein the anti-Fc receptor Ig is an anti-FcyR Ig, an anti-FcaR Ig or an anti-FcaR Ig, optionally wherein:(i) the anti-FcyR Ig is an anti-FcyR! Ig, an anti-FcyRIIA Ig, an anti-FcyRIIB Ig, an anti- FcyRIIIA Ig, or an anti-FcyRIIIB Ig;(ii) the anti-FcaR Ig is an anti-FcaRI Ig; and / or(iii)the anti-FcaR Ig is an anti-FcaRI Ig or an anti-FcsRII Ig.

20. The method of claim 1, wherein the sample is serum, plasma, a plasma fraction, a purified or partially purified plasma or plasma fraction thereof, a fermentation broth or a purified or partially purified fermentation broth thereof, a cell culture harvest or a purified or partially purified cell culture harvest thereof or a protein suspension, optionally wherein the plasma or plasma fraction is selected the group consisting of a human blood plasma sample, an IgG intermediate product, an intravenous immunoglobulin G (IVIG) product, a subcutaneous immunoglobulin G(SCIG) product, a cryo-rich plasma, a cryo- poor plasma, a Supernatant I (SN I), a Cohn Fraction II (Fr II), a Cohn Fraction II+III (Fr II+III), a Cohn Fraction (I+)II+III ((FrI+)II+III), a Kistler / Nitschmann Precipitate A (KN A), a Kistler / Nitschmann Precipitate B (KN B), a Kistler / Nitschmann Precipitate of Supernatant B (KN B+l), an IgG depleted intermediate product, a purified IgG product, a purified formulated IgG product, a formulated plasma fractionation product and combinations thereof.

21. The method of claim 19, wherein the method further comprises:(i) administering the sample to the subject if the anti-FcsRI Ig is present in the sample at a concentration of less than 500 pg / mL;(ii) including the plasma sample or fraction thereof into a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of less than 500 pg / mL; or(iii) excluding the plasma sample or fraction thereof from a pooled blood plasma sample if the anti-FcsRI Ig is present in the sample at a concentration of greater than 500 pg / mL.

22. The method of claim 19, wherein the anti-FcsRI Ig in the sample activates a FcsRI mediated signaling pathway if administered to a subject, optionally wherein anti-FcsRI Ig at a concentration of greater than 500 pg / mL activates a FcsRI mediated signaling pathway if administered to a subject, and optionally wherein activation of the FcsRI mediated signaling pathway induces basophil activation, mast cell activation, basophil degranulation and / or mast cell degranulation.

23. The method of claim 19, wherein the anti-FcsRI Ig is an anti-FcsRI IgG, an anti-FcsRI IgE, an anti-FcsRI IgM and / or an anti-FcsRI IgA, optionally wherein the anti-FcsRI IgG is an IgGl, an IgG2, an IgG3 or an IgG4.

24. The method of claim 1, wherein the method further comprises diluting the sample in an assay solution prior to contacting with the first binding moiety and the second binding moiety, optionally wherein the sample is diluted in the assay solution at a ratio of between 1 : 10 to 1 :20 and / or the sample is in the assay solution at a concentration of 10% (v / v).

25. A method of detecting anti-Fc epsilon receptor I (FcsRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, wherein the anti-FcsRI Ig is capable of inducing oligomerization and / or cross-linking of FcsRI, the method comprising:(i) contacting the sample comprising the anti-FcsRI Ig to a first binding moiety and a second binding moiety, wherein: a. the first binding moiety comprises: i. a first FcsRI a-chain or subunit thereof that specifically binds to a first binding domain of the anti-FcsRI Ig;ii. a first nucleic acid tag comprising a biotinylated tail; and iii. a first nucleic acid label; and b. the second binding moiety comprises: i. a second FcsRI a-chain or subunit thereof that specifically binds to a second binding domain of the anti-FcsRI Ig, ii. a second nucleic acid tag comprising an A, a T, a G and / or a C rich tail sequence; and iii. a second nucleic acid label, wherein the first and second binding moieties specifically bind to the anti-FcsRI Ig in the sample to thereby form an immunocomplex;(ii) contacting the immunocomplex to a first capture probe immobilized on a first solid matrix, wherein the first capture probe binds the first nucleic acid tag;(iii)generating a nucleic acid reporter from the immunocomplex based on proximity between the first nucleic acid label and the second nucleic acid label; and(iv)detecting the reporter, wherein the presence of the reporter is indicative of the presence of the anti-FcsRI Ig in the sample, optionally wherein prior to contacting the immunocomplex to the first capture probe immobilized on the first solid matrix, the method comprises:(iv) contacting the immunocomplex to a second capture probe immobilized on a second solid matrix, wherein the second capture probe binds the second nucleic acid tag;(v) washing the second solid matrix to remove unbound immunocomplex; and(vi)releasing the bound immunocomplex from the second immobilized capture probe.

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