Methods for identifying immunoglobulins associated with adverse reactions

Abstract

This disclosure relates to methods for identifying and reducing anti-Fc-epsilon receptor I (FcεRI) immunoglobulin (Ig) associated with adverse reactions from plasma or fractions thereof, and to the formulation and use of such anti-FcεRI Ig-reduced plasma protein products.

Description

[Technical Field]

[0001] This disclosure relates to methods for identifying and reducing anti-Fc-epsilon receptor I (FcεRI) immunoglobulin (Ig) associated with adverse reactions from plasma or fractions thereof, and to the formulation and use of such anti-FcεRI Ig-reduced plasma protein products. [Background technology]

[0002] Immunoglobulins (Ig) are one of the most abundant proteins in plasma and are responsible for complement activation, opsonization, and toxin neutralization. Plasma-derived Ig has become a major plasma product, and global demand is increasing. Both hyperimmune (or "specific") and normal (or "non-specific") human Ig products are primarily composed of IgG. Purified IgG from human plasma is used for the prophylactic prevention of infections in immunocompromised patients, replacement therapy for antibody deficiencies in patients, and the treatment of immunocompromised, inflammatory and autoimmune diseases, as well as conditions associated with acute infections in patients. A list of FDA-approved Ig is available at http: / / www.fda.gov / vaccines-blood-biologics / approved-blood-products / immune-globulins.

[0003] Currently, several commercially available intravenous IG (IVIG) products (typically 5% or 10% (w / v) stabilized solutions) include Privigen® (CSL Behring), Flebogamma® (Grifols), Gamunex®-C (Grifols), Gammagard® (Takeda), and Octagam® (Octapharma). More recently, subcutaneous IG (SCIG) administration has become available. Commercial SCIG products (typically 10%, 16.5%, or 20% (w / v) stabilized solutions) include Hizentra® (CSL Behring), Gamunex®-C (Grifols), Xembify® (Grifols), Cutaquig® (Octapharma), and Cuvitru® (Takeda). Other Ig products are administered intramuscularly (IMIG).

[0004] Immunoglobulin therapy is effective and generally well-tolerated, but various adverse effects have been reported over the years. Most of the events associated with receiving immunoglobulin products, such as rash, flushing, headache, malaise, fever, chills, fatigue, and lethargy, are transient and mild. However, some side effects, including immediate-type hypersensitivity reactions (including urticaria and pruritus), thrombosis, arrhythmias, aseptic meningitis, and hemolytic anemia, are serious. These adverse effects vary in severity and are associated with specific immunoglobulin preparations and individual patient sensitivity. While patient risk factors can be assessed, and adverse effects can be minimized by reducing the infusion rate and switching between IVIG and SCIG, it remains unclear which components of plasma and Ig products are causing these adverse reactions, and sometimes an increased frequency of adverse events can lead to the retrieval of Ig products from circulation. In fact, an excessive number of hypersensitivity reactions involving Ig in a particular lot can occur accidentally and unpredictably, leading to the isolation of the manufacturing intermediate stage, which can adversely impact the recall of the product lot and the supply of Ig (Product Safety Update; International blood / plasma news, p100, February 2023).

[0005] Therefore, it will be apparent to those skilled in the art that there is a need in the art for methods to identify and reduce components from plasma that cause adverse reactions in subjects from plasma samples or fractions thereof. [Overview of the project]

[0006] This disclosure is based on the inventors' identification of immunoglobulin (Ig) associated with adverse reactions in subjects. In particular, through statistical analysis of plasma donations, the inventors identified a single plasma donor associated with hypersensitivity reactions. Subsequently, the inventors found that plasma from this donor was able to induce degranulation of both mast cells and basophils via activation of the Fc epsilon receptor I (FcεRI) mediated pathway. The inventors further identified that the activation of the FcεRI mediated pathway was driven by immunoglobulin (Ig) G. Somewhat surprisingly, purified IgG products derived from the plasma of donors associated with hypersensitivity reactions did not clearly induce degranulation of mast cells or basophils in vitro via activation of the FcεRI mediated pathway. Only by identifying the individual plasma donors associated with hypersensitivity reactions could the inventors determine why the purified IgG products caused adverse reactions in subjects.

[0007] Therefore, the present inventors' findings provide a basis for a method for identifying anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof. The disclosure also provides a method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof. The disclosure further provides a method for screening a plasma sample or fraction thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig).

[0008] This disclosure provides a method comprising contacting a plasma sample or fraction thereof with a reagent that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the plasma sample or fraction thereof, and detecting the binding of the reagent.

[0009] In one example, the reagent directly binds to anti-FcεRI Ig in the plasma sample or a fraction thereof.

[0010] In one example, the reagent indirectly binds to anti-FcεRI Ig in the plasma sample or its fraction.

[0011] In one example, the reagent is a ligand containing FcεRI or a fragment or epitope thereof. For example, a fragment of FcεRI contains the α chain of FcεRI. In another example, the FcεRI fragment is a functional fragment. For example, a functional FcεRI fragment is a fragment of FcεRI that activates an FcεRI-mediated signaling pathway when bound to anti-FcεRI Ig.

[0012] In one example, the method involves contacting a plasma sample or fraction thereof with a ligand containing FcεRI or a fragment or epitope that binds to anti-FcεRI Ig in the plasma sample or fraction thereof. For example, the binding of anti-FcεRI Ig in the plasma sample or fraction thereof to FcεRI or a fragment or epitope results in the formation of a complex. In one example, the method further includes detecting the complex, the presence of which indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0013] This disclosure provides a method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, the method being: (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) detecting a complex, the presence of which indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0014] This disclosure also provides a method for screening plasma samples or fractions thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), the method being: (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) detecting a complex, the presence of which indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0015] The present disclosure further provides a method for identifying a plasma sample or a fraction thereof suitable for administration to a subject, the method comprising: (i) contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex; (ii) detecting the complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in the plasma sample or the fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or the fraction thereof indicates that the plasma sample or the fraction thereof is not suitable for administration to the subject.

[0016] The present disclosure provides a method for screening a plasma sample or a fraction thereof to determine suitability for administration to a subject, the method comprising: (i) contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex; (ii) detecting the complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in the plasma sample or the fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or the fraction thereof indicates that the plasma sample or the fraction thereof is not suitable for administration to the subject.

[0017] The present disclosure further provides a method for identifying a plasma sample or a fraction thereof for use in manufacturing an immunoglobulin (Ig) preparation for administration to a subject, the method comprising: (i) contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) Ig in the sample, thereby forming a complex; (ii) detecting the complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in the plasma sample or the fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or the fraction thereof indicates that the plasma sample or the fraction thereof is not suitable for use in manufacturing an Ig preparation for administration to the subject.

[0018] This disclosure also provides a method for screening plasma samples or fractions thereof for use in preparing immunoglobulin (Ig) preparations for administration to a subject, the method being: (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI)Ig in the sample, thereby forming a complex, (ii) detecting a complex, the presence of which indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0019] This disclosure provides a method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, the method being: (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) detecting the complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0020] This disclosure provides a method for screening plasma samples or fractions thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), the method being: (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) detecting the complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0021] This disclosure provides a method for identifying a plasma sample or fraction thereof suitable for administration to a subject, and the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) Detection of the complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is not suitable for administration to the subject.

[0022] This disclosure provides a method for screening plasma samples or fractions thereof to determine their suitability for administration to a subject, and the method is: (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin Ig in the sample, thereby forming a complex, (ii) Detection of the complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is not suitable for administration to the subject.

[0023] This disclosure provides a method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, the method being: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) detecting a label, the presence of which a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0024] This disclosure provides a method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, the method being: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) detecting a label, the presence of which a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0025] This disclosure provides a method for screening plasma samples or fractions thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), the method being: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) detecting a label, the presence of which a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0026] This disclosure provides a method for screening plasma samples or fractions thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), the method being: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) detecting a label, the presence of which a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0027] This disclosure provides a method for identifying a plasma sample or fraction thereof suitable for administration to a subject, and the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is not suitable for administration to a subject.

[0028] This disclosure provides a method for identifying a plasma sample or fraction thereof suitable for administration to a subject, and the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction, and the presence of anti-FcεRI Ig in the plasma sample or fraction indicates that the plasma sample or fraction is not suitable for administration to a subject.

[0029] This disclosure provides a method for screening plasma samples to determine their suitability for administration to a subject, and the method is: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is not suitable for administration to a subject.

[0030] This disclosure provides a method for screening plasma samples to determine their suitability for administration to a subject, and the method is: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction, and the presence of anti-FcεRI Ig in the plasma sample or fraction indicates that the plasma sample or fraction is not suitable for administration to a subject.

[0031] This disclosure provides a method for identifying suitable subjects for plasma donation for the manufacture of immunoglobulin (Ig) preparations, and the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the subject is not suitable for plasma donation for the manufacture of an Ig preparation.

[0032] This disclosure provides a method for screening subjects suitable for plasma donation for the manufacture of immunoglobulin (Ig) preparations, and the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the subject is not suitable for plasma donation for the manufacture of an Ig preparation.

[0033] This disclosure provides a method for identifying suitable subjects for plasma donation for the manufacture of immunoglobulin (Ig) preparations, and the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) detecting the complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the subject is not suitable for plasma donation for the manufacture of an Ig preparation.

[0034] This disclosure provides a method for screening subjects suitable for plasma donation for the manufacture of immunoglobulin (Ig) preparations, and the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) detecting the complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the subject is not suitable for plasma donation for the manufacture of an Ig preparation.

[0035] In one example, the ligand includes FcεRI or a fragment or epitope thereof. For example, a fragment of FcεRI includes the α chain of FcεRI. In another example, the FcεRI fragment is a functional fragment. For example, a functional FcεRI fragment is a fragment of FcεRI that activates an FcεRI-mediated signaling pathway when bound to anti-FcεRI Ig.

[0036] In some cases, anti-FcεRI Ig binds to FcεRI epitopes and activates FcεRI-mediated signaling pathways.

[0037] In one example, anti-FcεRI Ig binds to the α-chain of FcεRI and activates the FcεRI-mediated signaling pathway.

[0038] In one example, anti-FcεRI Ig binds to the epitope of FcεRI and activates another immunological signaling pathway. For example, anti-FcεRI Ig binds to the epitope of FcεRI and activates an IgE-mediated signaling pathway. In another example, anti-FcεRI Ig binds to the epitope of FcεRI and activates an IgG-mediated signaling pathway. In one example, anti-FcεRI Ig binds to the epitope of FcεRI, an IgE-mediated signaling pathway, and another immunological signaling pathway. In another example, anti-FcεRI Ig binds to the epitope of FcεRI, an IgG-mediated signaling pathway, and another immunological signaling pathway. In several examples, anti-FcεRI Ig binds to the epitope of FcεRI and activates both an IgE-mediated and an IgG-mediated signaling pathway. In one example, anti-FcεRI Ig binds to the epitope of FcεRI and activates the FcεRI-mediated signaling pathway, the IgE-mediated signaling pathway, and the IgG-mediated signaling pathway. In another example, anti-FcεRI Ig binds to the epitope of FcεRI and activates the FcεRI-mediated signaling pathway and the IgE-mediated signaling pathway. In yet another example, anti-FcεRI Ig binds to the epitope of FcεRI and activates the FcεRI-mediated signaling pathway and the IgG-mediated signaling pathway.

[0039] In one example, activation of the FcεRI-mediated signaling pathway induces activation and / or degranulation of basophils and / or mast cells. In another example, activation of the FcεRI-mediated signaling pathway induces basophil activation. In yet another example, activation of the FcεRI-mediated signaling pathway induces basophil degranulation. In yet another example, activation of the FcεRI-mediated signaling pathway induces activation and degranulation of basophils. In yet another example, activation of the FcεRI-mediated signaling pathway induces activation of mast cells. In yet another example, activation of the FcεRI-mediated signaling pathway induces degranulation of mast cells. In one example, activation of the FcεRI-mediated signaling pathway induces activation and degranulation of mast cells. In yet another example, activation of the FcεRI-mediated signaling pathway induces activation and degranulation of both basophils and mast cells.

[0040] The disclosure also provides a method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, the method comprising detecting activation and / or degranulation of basophils and / or mast cells induced by the plasma sample or fraction thereof, the activation and / or degranulation of basophils and / or mast cells indicating the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0041] This disclosure provides a method for screening a plasma sample or fraction thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), the method comprising detecting activation and / or degranulation of basophils and / or mast cells induced by the plasma sample or fraction thereof, the activation and / or degranulation of basophils and / or mast cells indicating the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0042] The Disclosure also provides a method for screening plasma samples or fractions thereof for use in preparing immunoglobulin (Ig) preparations for administration to a subject, the method comprising detecting activation and / or degranulation of basophils and / or mast cells induced by the plasma sample or fraction thereof, the activation and / or degranulation of basophils and / or mast cells indicating the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicating that the plasma sample or fraction thereof is unsuitable for use in preparing Ig preparations for administration to a subject.

[0043] The Disclosure further provides a method for identifying a plasma sample or fraction thereof suitable for administration to a subject, the method comprising detecting activation and / or degranulation of basophils and / or mast cells induced by the plasma sample or fraction thereof, the activation and / or degranulation of basophils and / or mast cells indicating the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicating that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0044] The Disclosure further provides a method for identifying a plasma sample or fraction thereof for use in preparing an immunoglobulin (Ig) preparation for administration to a subject, the method comprising detecting activation and / or degranulation of basophils and / or mast cells induced by the plasma sample or fraction thereof, the percentage of basophil and / or mast cell activation and / or degranulation indicating the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicating that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0045] In one example, anti-FcεRI Ig is anti-FcεRI IgG and / or anti-FcεRI IgE. In another example, anti-FcεRI Ig is anti-FcεRI IgG and anti-FcεRI IgE. In yet another example, anti-FcεRI Ig is anti-FcεRI IgG. In yet another example, anti-FcεRI Ig is anti-FcεRI IgE.

[0046] In one example, anti-FcεRI Ig induces a hypersensitivity reaction in a subject.

[0047] In one example, the ligand is immobilized on a solid surface.

[0048] In another example, the method further includes immobilizing the FcεRI ligand onto a solid surface.

[0049] In one example, the method further comprises a protein complex in contact with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig, and the detection protein contains a detectable label.

[0050] In one example, the method further comprises a protein complex in contact with a detection protein containing an antibody variable region, the detection protein being bound to anti-FcεRI Ig, and the detection protein being brought into contact with an antibody containing a label that can be bound to it and detected.

[0051] In one example, the detected protein is anti-IgG and / or anti-IgE. In another example, the detected protein is anti-IgG and anti-IgE. In yet another example, the detected protein is anti-IgG. In yet another example, the detected protein is anti-IgE.

[0052] For example, detectable labels are selected from the group consisting of radioactive labels, enzymes, fluorescent labels, luminescent labels, bioluminescent labels, magnetic labels, prosthetic groups, and contrast agents.

[0053] Those skilled in the art will understand from the disclosure herein that, in a method for screening and / or identifying plasma samples or fractions suitable for administration and / or use in the preparation of the Ig preparations described herein, a plasma sample or fraction is unsuitable for administration or use if anti-FcεRI IgG is present in a detectable amount. For example, the presence of a detectable amount of anti-FcεRI Ig in a plasma sample or fraction indicates that the plasma sample or fraction is unsuitable for administration to a subject. In another example, the presence of a detectable amount of anti-FcεRI Ig in a plasma sample or fraction indicates that the plasma sample or fraction is unsuitable for use in the preparation of an Ig preparation for administration to a subject.

[0054] Methods for detecting the presence of anti-FcεRI Ig will be apparent to those skilled in the art and / or will be described herein. For example, the presence of a detectable amount of anti-FcεRI Ig can be determined directly by determining the level of anti-FcεRI Ig in a plasma sample or fraction thereof. In another example, the presence of a detectable amount of anti-FcεRI Ig can be determined indirectly by determining the rate of activation and / or degranulation of basophils and / or mast cells induced by a plasma sample or fraction thereof.

[0055] In one example, the method involves determining the level of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0056] For example, the method includes determining the level of anti-FcεRI IgG in a plasma sample or fraction thereof. For example, the method includes determining a detectable amount of anti-FcεRI IgG in a plasma sample or fraction thereof. For example, the detectable amount of anti-FcεRI IgG in a plasma sample or fraction thereof is at least 40 μg / mL. For example, the detectable amount of anti-FcεRI IgG in a plasma sample or fraction thereof is at least 45 μg / mL. For example, the detectable amount of anti-FcεRI IgG in a plasma sample or fraction thereof is at least 50 μg / mL. For example, if anti-FcεRI IgG is present in a plasma sample or fraction thereof at a concentration of at least 40 μg / mL (i.e., in a detectable amount), the plasma sample or fraction thereof is not suitable for administration to a subject or for use in preparing Ig preparations for administration to a subject. For example, if anti-FcεRI IgG is present in a plasma sample or fraction at a concentration of less than 40 μg / mL (i.e., not in a detectable amount), the plasma sample or fraction is suitable for administration to a subject or for use in preparing an Ig preparation for administration to a subject.

[0057] For example, the method includes determining the level of anti-FcεRI IgE in a plasma sample or fraction thereof. For instance, the method includes determining a detectable amount of anti-FcεRI IgE in a plasma sample or fraction thereof. For example, the detectable amount of anti-FcεRI IgE in a plasma sample or fraction thereof is at least 0.1 kU / L. For example, the detectable amount of anti-FcεRI IgE in a plasma sample or fraction thereof is at least 0.5 kU / L. For example, the detectable amount of anti-FcεRI IgE in a plasma sample or fraction thereof is at least 1 kU / L. For example, if anti-FcεRI IgE is present in a plasma sample or fraction thereof at a concentration of at least 0.1 kU / L (i.e., in a detectable amount), the plasma sample or fraction thereof is not suitable for administration to a subject or for use in preparing Ig preparations for administration to a subject. For example, if anti-FcεRI IgE is present in a plasma sample or fraction at a concentration of less than 0.1 kU / L (i.e., not detectable), the plasma sample or fraction is suitable for administration to a subject or for use in preparing Ig preparations for administration to a subject.

[0058] In one example, the method further includes determining the rate of basophil and / or mast cell activation and / or degranulation induced by a plasma sample or fraction thereof. In another example, the method further includes determining the rate of basophil and mast cell activation and / or degranulation induced by a plasma sample or fraction thereof. In yet another example, the method further includes determining the rate of basophil activation and / or degranulation induced by a plasma sample or fraction thereof. In yet another example, the method further includes determining the rate of basophil activation and / or degranulation induced by a plasma sample or fraction thereof.

[0059] In one example, the rate of basophil and / or mast cell activation and / or degranulation after exposure to a plasma sample or fraction thereof is determined by determining the percentage of basophils and / or mast cells expressing one or more markers selected from the group consisting of CD63, CD203c, CD107a, and combinations thereof. In one example, the rate of basophil and mast cell activation after exposure to a plasma sample or fraction thereof is determined by determining the percentage of basophils and mast cells expressing one or more markers selected from the group consisting of CD63, CD203c, CD107a, and combinations thereof. In one example, the rate of basophil activation after exposure to a plasma sample or fraction thereof is determined by determining the percentage of basophils expressing one or more markers selected from the group consisting of CD63, CD203c, and combinations thereof.

[0060] In one example, the rate of mast cell activation after exposure to a plasma sample or fraction thereof is determined by determining the percentage of mast cells expressing CD107a.

[0061] For example, the method includes determining the percentage of basophils that express CD63 after exposure to a plasma sample or fraction thereof. For instance, the method includes determining a detectable amount of anti-FcεRI Ig in a plasma sample or fraction thereof by determining the percentage of basophils that express CD63 after exposure to a plasma sample or fraction thereof. For example, the percentage of basophils that express CD63 after exposure to a plasma sample or fraction thereof is greater than 5%. For example, the percentage of basophils that express CD63 is relative to the total percentage of basophils exposed to the plasma sample or fraction thereof. For example, if at least 5% of the population of basophils express CD63 after exposure to a plasma sample or fraction thereof, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction thereof.

[0062] For example, the method includes determining the percentage of basophils that express CD203c after exposure to a plasma sample or fraction thereof. For instance, the method includes determining a detectable amount of anti-FcεRI Ig in a plasma sample or fraction thereof by determining the percentage of basophils that express CD203c after exposure to a plasma sample or fraction thereof. For example, the percentage of basophils that express CD203c after exposure to a plasma sample or fraction thereof is greater than 5%. For example, the percentage of basophils that express CD203c is relative to the total percentage of basophils exposed to the plasma sample or fraction thereof. For example, if at least 5% of the population of basophils express CD203c after exposure to a plasma sample or fraction thereof, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction thereof.

[0063] For example, the method includes determining the percentage of mast cells expressing CD107a after exposure to a plasma sample or fraction thereof. For instance, the method includes determining a detectable amount of anti-FcεRI Ig in a plasma sample or fraction thereof by determining the percentage of mast cells expressing CD107a after exposure to a plasma sample or fraction thereof. For example, the percentage of mast cells expressing CD107a after exposure to a plasma sample or fraction thereof is greater than 5%. For example, the percentage of mast cells expressing CD107a is relative to the total percentage of mast cells exposed to the plasma sample or fraction thereof. For example, if at least 5% of the population of mast cells express CD107a after exposure to a plasma sample or fraction thereof, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction thereof.

[0064] In one example, the plasma sample or fraction thereof is selected from the group consisting of human blood plasma sample, IgG intermediate product, intravenous immunoglobulin G (IVIG), subcutaneous immunoglobulin G (SCIG), cryo-rich plasma, decryoplasma, supernatant I (SN I), Cohn fraction II (Fr II), Cohn fraction II+III (Fr II+III), Cohn fraction I+II+III (Fr I+II+III), Kistler / Nitzjemann precipitate A (KN A), Kistler / Nitzjemann precipitate B (KN B), Kistler / Nitzjemann precipitate of supernatant B (KN B+1), and combinations thereof.

[0065] In one example, the plasma sample or fraction thereof is a human blood plasma sample from one or more subjects.

[0066] In one example, the method further includes administering a plasma sample or fraction thereof if anti-FcεRI Ig is not present. For example, anti-FcεRI Ig is not present in a detectable amount.

[0067] For example, the method further includes including a plasma sample or fraction thereof in a pooled blood plasma sample if anti-FcεRI Ig is not present. For instance, if anti-FcεRI Ig is not present in a detectable amount, the plasma sample or fraction thereof is included in the pooled blood plasma sample.

[0068] In one example, the method further includes excluding the plasma sample or fraction thereof from the pooled blood plasma sample if anti-FcεRI Ig is present. For example, if anti-FcεRI Ig is present in a detectable amount, the plasma sample or fraction thereof is excluded from the pooled blood plasma sample.

[0069] The inventors' findings also provide a basis for a method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation from a plasma sample or fraction thereof, the method comprising detecting anti-FcεRI Ig in a plasma sample or fraction thereof according to any method described herein, and producing an anti-FcεRI Ig depletion preparation.

[0070] This disclosure also provides a method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation from a plasma sample or fraction thereof, the method comprising conjugating anti-FcεRI Ig to a chromatographic resin containing a ligand that binds to anti-FcεRI Ig in a plasma sample or fraction thereof, and collecting the anti-FcεRI Ig depletion preparation. This disclosure also provides a method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation from a plasma sample or fraction thereof, the method comprising, (i) Loading a plasma sample or fraction thereof onto a chromatography resin containing a ligand that binds to anti-FcεRI Ig in the plasma sample or fraction thereof, (ii) Collecting anti-FcεRI Ig reduction preparations, which includes

[0071] For example, a chromatography resin is an affinity chromatography resin. For instance, a continuous affinity chromatography resin.

[0072] This disclosure provides a method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation from a plasma sample or fraction thereof, the method being: (i) Loading a plasma sample or fraction thereof onto an affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig in the plasma sample or fraction thereof, (ii) Collecting anti-FcεRI Ig reduction preparations, which includes

[0073] In one example, the ligand is immobilized on a matrix of chromatography resin.

[0074] In one example, the ligand includes FcεRI or a fragment or epitope thereof. For example, a fragment of FcεRI includes the α chain of FcεRI. In another example, the FcεRI fragment is a functional fragment. For example, a functional FcεRI fragment is a fragment of FcεRI that activates an FcεRI-mediated signaling pathway when bound to anti-FcεRI Ig.

[0075] In one example, the method further includes detecting the presence of anti-FcεRI Ig in a plasma sample or fraction thereof by performing the detection method described herein.

[0076] In one example, an anti-FcεRI Ig-reducing preparation induces reduced activation of the FcεRI-mediated signaling pathway compared to a preparation in which anti-FcεRI Ig levels are not reduced. In another example, an anti-FcεRI Ig-reducing preparation does not induce activation of the FcεRI-mediated signaling pathway.

[0077] In one example, an anti-FcεRI Ig-reduced preparation induces reduced basophil and mast cell activation and / or degranulation compared to a preparation in which anti-FcεRI Ig levels are not reduced. In another example, an anti-FcεRI Ig-reduced preparation induces reduced basophil activation compared to a preparation in which anti-FcεRI Ig levels are not reduced. In yet another example, an anti-FcεRI Ig-reduced preparation induces reduced mast cell activation compared to a preparation in which anti-FcεRI Ig levels are not reduced. In yet another example, an anti-FcεRI Ig-reduced preparation does not induce basophil and / or mast cell activation and / or degranulation.

[0078] In one example, the activation and / or degranulation of basophils and / or mast cells after exposure to an anti-FcεRI Ig-reducing preparation is determined by the method described herein. For example, the method includes determining the percentage of basophils 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 activation of basophils after exposure to an anti-FcεRI Ig-reducing preparation is determined by determining the percentage of basophils expressing one or more markers selected from the group consisting of CD63, CD203c, and combinations thereof. In yet another example, the activation of mast cells after exposure to an anti-FcεRI Ig-reducing preparation is determined by determining the percentage of mast cells expressing CD107a.

[0079] In one example, the method includes determining the percentage of basophils expressing CD63 after exposure to an anti-FcεRI Ig depletion preparation. For example, the method includes determining whether a detectable amount of anti-FcεRI Ig is present in the anti-FcεRI Ig depletion preparation by determining the percentage of basophils expressing CD63 after exposure to the anti-FcεRI Ig depletion preparation. In one example, the percentage of basophils expressing CD63 after exposure to the anti-FcεRI Ig depletion preparation is less than 5%. For example, the percentage of basophils expressing CD63 is relative to the total percentage of basophils exposed to the anti-FcεRI Ig depletion preparation. In one example, if at least 5% of the basophil population expresses CD63 after exposure to the anti-FcεRI Ig depletion preparation, then a detectable amount of anti-FcεRI Ig is present in the anti-FcεRI Ig depletion preparation. In one example, if less than 5% of the basophil population expresses CD63 after exposure to an anti-FcεRI Ig-reducing preparation, then a detectable amount of anti-FcεRI Ig is not present in the anti-FcεRI Ig-reducing preparation.

[0080] In one example, the method includes determining the percentage of basophils expressing CD203c after exposure to an anti-FcεRI Ig depletion preparation. For example, the method includes determining whether a detectable amount of anti-FcεRI Ig is present in the anti-FcεRI Ig depletion preparation by determining the percentage of basophils expressing CD203c after exposure to the anti-FcεRI Ig depletion preparation. In one example, the percentage of basophils expressing CD203c after exposure to the anti-FcεRI Ig depletion preparation is less than 5%. For example, the percentage of basophils expressing CD203c is relative to the total percentage of basophils exposed to the anti-FcεRI Ig depletion preparation. In one example, if at least 5% of the basophil population expresses CD203c after exposure to the anti-FcεRI Ig depletion preparation, then a detectable amount of anti-FcεRI Ig is present in the anti-FcεRI Ig depletion preparation. In one example, if less than 5% of the basophil population expresses CD203c after exposure to an anti-FcεRI Ig depletion preparation, then a detectable amount of anti-FcεRI Ig is not present in the anti-FcεRI Ig depletion preparation.

[0081] In one example, the method includes determining the percentage of mast cells expressing CD107a after exposure to an anti-FcεRI Ig-reducing preparation. For example, the method includes determining whether a detectable amount of anti-FcεRI Ig is present in the anti-FcεRI Ig-reducing preparation by determining the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation. In one example, the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 5%. For example, the percentage of mast cells expressing CD107a is relative to the total percentage of mast cells exposed to the anti-FcεRI Ig-reducing preparation. In one example, if at least 5% of the population of mast cells express CD107a after exposure to the anti-FcεRI Ig-reducing preparation, then a detectable amount of anti-FcεRI Ig is present in the anti-FcεRI Ig-reducing preparation. For example, if less than 5% of the mast cell population expresses CD107a after exposure to an anti-FcεRI Ig-reducing preparation, then a detectable amount of anti-FcεRI Ig is not present in the anti-FcεRI Ig-reducing preparation.

[0082] For example, the anti-FcεRI Ig reduction preparation contains less than 40 μg / mL of anti-FcεRI IgG and less than 0.1 kU / L of anti-FcεRI IgE.

[0083] In one example, the method includes determining the level of anti-FcεRI IgG in an anti-FcεRI Ig reduction preparation. For example, the method includes determining whether a detectable amount of anti-FcεRI IgG is present in the anti-FcεRI Ig reduction preparation. In one example, the detectable amount of anti-FcεRI IgG in the anti-FcεRI Ig reduction preparation is at least 40 μg / mL. In another example, the anti-FcεRI Ig reduction preparation contains less than 40 μg / mL of anti-FcεRI IgG. In one example, the level of anti-FcεRI IgG in the anti-FcεRI Ig reduction preparation is less than 35 μg / mL. In one example, the level of anti-FcεRI IgG in the anti-FcεRI Ig reduction preparation is less than 30 μg / mL. In one example, the level of anti-FcεRI IgG in the anti-FcεRI Ig reduction preparation is less than 35 μg / mL. In one example, the level of anti-FcεRI IgG in the anti-FcεRI Ig-reducing preparation is less than 20 μg / mL. In another example, the level of anti-FcεRI IgG in the anti-FcεRI Ig-reducing preparation is less than 15 μg / mL. In another example, the level of anti-FcεRI IgG in the anti-FcεRI Ig-reducing preparation is less than 10 μg / mL. In another example, the level of anti-FcεRI IgG in the anti-FcεRI Ig-reducing preparation is less than 5 μg / mL. In one example, if anti-FcεRI IgG is present in the anti-FcεRI Ig-reducing preparation at a concentration of less than 40 μg / mL (i.e., not detectable), the anti-FcεRI Ig-reducing preparation is suitable for administration to the subject.

[0084] For example, the method includes determining the level of anti-FcεRI IgE in an anti-FcεRI Ig depletion preparation. For instance, the method includes determining whether a detectable amount of anti-FcεRI IgE is present in the anti-FcεRI Ig depletion preparation. For example, the detectable amount of anti-FcεRI IgE in the anti-FcεRI Ig depletion preparation is at least 0.1 kU / L. For example, the anti-FcεRI Ig depletion preparation contains less than 0.1 kU / L of anti-FcεRI IgE. For example, the level of anti-FcεRI IgE in the anti-FcεRI Ig depletion preparation is less than 0.01 kU / L. For example, the level of anti-FcεRI IgE in the anti-FcεRI Ig depletion preparation is less than 0.05 kU / L. For example, the level of anti-FcεRI IgE in the anti-FcεRI Ig depletion preparation is less than 0.001 kU / L. In one example, the level of anti-FcεRI IgE in the anti-FcεRI Ig-reducing preparation is less than 0.005 kU / L. In another example, if anti-FcεRI IgE is present in the anti-FcεRI Ig-reducing preparation at a concentration of less than 0.1 kU / L (i.e., not detectable), the anti-FcεRI Ig-reducing preparation is suitable for administration to the subject.

[0085] For example, an anti-FcεRI Ig-reducing preparation is an anti-FcεRI Ig-reducing IgG preparation.

[0086] In one example, the affinity chromatography resin further comprises blood group A antigen and blood group B antigen immobilized on the matrix of the affinity chromatography resin. In another example, the affinity chromatography resin further comprises blood group A antigen. In yet another example, the affinity chromatography resin further comprises blood group B antigen.

[0087] In one example, an affinity chromatography resin is a continuous affinity chromatography resin. For instance, a continuous affinity chromatography resin contains a ligand that binds to anti-FcεRI Ig.

[0088] In one example, the affinity chromatography resin is a continuous affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, blood group A antigen, and blood group B antigen.

[0089] In one example, the affinity chromatography resin is a continuous affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig and a blood group A antigen, immobilized on the matrix of the affinity chromatography resin.

[0090] In one example, the affinity chromatography resin is a continuous affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig and a blood group B antigen, immobilized on the affinity chromatography resin matrix.

[0091] This disclosure provides affinity chromatography resins comprising ligands that bind to anti-FcεRI Ig, and optionally, blood group A antigens and / or blood group B antigens. For example, this disclosure provides serial affinity chromatography resins comprising ligands that bind to anti-FcεRI Ig, and optionally, blood group A antigens and / or blood group B antigens.

[0092] For example, affinity chromatography is, (i) A first affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the first affinity chromatography resin, (ii) A continuous affinity chromatography comprising a second affinity chromatography resin containing blood type A antigen and blood type B antigen immobilized on a matrix of the second affinity chromatography resin.

[0093] For example, affinity chromatography is, (i) A first affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the first affinity chromatography resin, (ii) A continuous affinity chromatography comprising a second affinity chromatography resin containing a blood type A antigen immobilized on a matrix of the second affinity chromatography resin.

[0094] For example, affinity chromatography is, (i) A first affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the first affinity chromatography resin, (ii) A continuous affinity chromatography comprising a second affinity chromatography resin containing a blood group B antigen immobilized on the matrix of the second affinity chromatography resin.

[0095] For example, affinity chromatography is, (i) A first affinity chromatography resin comprising blood type A antigen and blood type B antigen immobilized on a matrix of the first affinity chromatography resin, (ii) A continuous affinity chromatography comprising a second affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the second affinity chromatography resin.

[0096] For example, affinity chromatography is, (i) A first affinity chromatography resin containing blood group A antigen immobilized on the matrix of the first affinity chromatography resin, (ii) A continuous affinity chromatography comprising a second affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the second affinity chromatography resin.

[0097] For example, affinity chromatography is, (i) A first affinity chromatography resin containing blood group B antigen immobilized on the matrix of the first affinity chromatography resin, (ii) A continuous affinity chromatography comprising a second affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the second affinity chromatography resin.

[0098] In one example, the ligand includes FcεRI or a fragment or epitope thereof. For example, a fragment of FcεRI includes the α chain of FcεRI. In another example, the FcεRI fragment is a functional fragment. For example, a functional FcεRI fragment is a fragment of FcεRI that activates an FcεRI-mediated signaling pathway when bound to anti-FcεRI Ig.

[0099] In one example, the method further includes one or more steps selected from the group consisting of ethanol precipitation, octanoid acid fractionation, ion exchange chromatography, virus inactivation, viral filtration, ultrafiltration / dialysis filtration, and combinations thereof.

[0100] The findings provide a basis for pharmaceutical compositions comprising polyclonal IgG, anti-FcεRI Ig-decreasing preparations, or anti-FcεRI Ig-decreasing IgG plasma preparations, as well as the use of these pharmaceutical compositions for treating, preventing, and / or delaying the progression of conditions in subjects (e.g., primary immunodeficiency, chronic inflammatory demyelinating polyneuropathy, and chronic immune thrombocytopenic purpura).

[0101] This disclosure provides a pharmaceutical composition comprising an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depleting IgG preparation manufactured by the method described herein.

[0102] For example, 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).

[0103] In one example, the anti-FcεRI Ig reduction preparation contains an undetectable amount of anti-FcεRI Ig. For example, the anti-FcεRI Ig reduction preparation contains less than 40 μg / mL of anti-FcεRI IgG and / or less than 0.1 kU / L of anti-FcεRI IgE. In one example, the anti-FcεRI Ig reduction preparation contains less than 40 μg / mL of anti-FcεRI IgG. In one example, the anti-FcεRI Ig reduction preparation contains less than 35 μg / mL of anti-FcεRI IgG. In one example, the anti-FcεRI Ig reduction preparation contains less than 30 μg / mL of anti-FcεRI IgG. In one example, the anti-FcεRI Ig reduction preparation contains less than 25 μg / mL of anti-FcεRI IgG. In one example, the anti-FcεRI Ig reduction preparation contains less than 20 μg / mL of anti-FcεRI IgG. In one example, the anti-FcεRI Ig-reducing preparation contains less than 15 μg / mL of anti-FcεRI IgG. In another example, the anti-FcεRI Ig-reducing preparation contains less than 10 μg / mL of anti-FcεRI IgG. In another example, the anti-FcεRI Ig-reducing preparation contains less than 5 μg / mL of anti-FcεRI IgG. In another example, the anti-FcεRI Ig-reducing preparation contains less than 0.1 kU / L of anti-FcεRI IgE. In another example, the anti-FcεRI Ig-reducing preparation contains less than 0.01 kU / L of anti-FcεRI IgE. In another example, the anti-FcεRI Ig-reducing preparation contains less than 0.005 kU / L of anti-FcεRI IgE. In another example, the anti-FcεRI Ig-reducing preparation contains less than 0.001 kU / L of anti-FcεRI IgE.

[0104] In one example, an anti-FcεRI Ig-reduced IgG preparation contains an undetectable amount of anti-FcεRI Ig. For example, an anti-FcεRI Ig-reduced IgG preparation contains less than 40 μg / mL of anti-FcεRI IgG and / or less than 0.1 kU / L of anti-FcεRI IgE. In one example, an anti-FcεRI Ig-reduced IgG preparation contains less than 40 μg / mL of anti-FcεRI IgG. In one example, an anti-FcεRI Ig-reduced IgG preparation contains less than 35 μg / mL of anti-FcεRI IgG. In one example, an anti-FcεRI Ig-reduced IgG preparation contains less than 30 μg / mL of anti-FcεRI IgG. In one example, an anti-FcεRI Ig-reduced IgG preparation contains less than 25 μg / mL of anti-FcεRI IgG. In one example, an anti-FcεRI Ig-reduced IgG preparation contains less than 20 μg / mL of anti-FcεRI IgG. In one example, the anti-FcεRI Ig-reducing IgG preparation contains less than 15 μg / mL of anti-FcεRI IgG. In another example, the anti-FcεRI Ig-reducing IgG preparation contains less than 10 μg / mL of anti-FcεRI IgG. In another example, the anti-FcεRI Ig-reducing IgG preparation contains less than 5 μg / mL of anti-FcεRI IgG. In another example, the anti-FcεRI Ig-reducing IgG preparation contains less than 0.1 kU / L of anti-FcεRI IgE. In another example, the anti-FcεRI Ig-reducing IgG preparation contains less than 0.01 kU / L of anti-FcεRI IgE. In another example, the anti-FcεRI Ig-reducing IgG preparation contains less than 0.005 kU / L of anti-FcεRI IgE. In another example, the anti-FcεRI Ig-reducing IgG preparation contains less than 0.001 kU / L of anti-FcεRI IgE.

[0105] In one example, anti-FcεRI Ig-reducing preparations do not induce activation of FcεRI-mediated signaling pathways. For example, anti-FcεRI Ig-reducing preparations do not induce activation and / or degranulation of basophils and / or mast cells. In another example, anti-FcεRI Ig-reducing preparations do not induce activation and / or degranulation of detectable amounts of basophils and / or mast cells.

[0106] It will be apparent from the disclosure herein that the activation and / or degranulation of basophils and / or mast cells is determined by determining the proportion of basophils expressing CD63 and / or CD203c and / or mast cells expressing CD107a. In one example, of the total population of basophils and / or mast cells exposed to the anti-FcεRI Ig-reduced IgG preparation, the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reduced IgG preparation is less than 5%, the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reduced IgG preparation is less than 5%, and / or the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reduced IgG preparation is less than 5%. In another example, the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparation. In further examples, the percentage of basophils expressing CD203c after exposure to anti-FcεRI Ig-reduced IgG preparations is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparations. In one example, the percentage of mast cells expressing CD107a after exposure to anti-FcεRI Ig-reduced IgG preparations is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reduced IgG preparations. In another example, the percentage of basophils expressing CD63 after exposure to anti-FcεRI Ig-reduced IgG preparations is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparations, and / or the percentage of basophils expressing CD203c after exposure to anti-FcεRI Ig-reduced IgG preparations is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparations. In one example, the proportion of basophils expressing CD63 after exposure to an anti-FcεRI Ig-reduced IgG preparation was less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparation, and / or the proportion of mast cells expressing CD107a after exposure to an anti-FcεRI Ig-reduced IgG preparation was less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reduced IgG preparation.In one example, the proportion of basophils expressing CD203c after exposure to an anti-FcεRI Ig-reduced IgG preparation was less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparation, and / or the proportion of mast cells expressing CD107a after exposure to an anti-FcεRI Ig-reduced IgG preparation was less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reduced IgG preparation. For example, the proportion of basophils expressing CD63 after exposure to an anti-FcεRI Ig-reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparation, the proportion of basophils expressing CD203c after exposure to an anti-FcεRI Ig-reduced IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reduced IgG preparation, and / or the proportion of mast cells expressing CD107a after exposure to an anti-FcεRI Ig-reduced IgG preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reduced IgG preparation.

[0107] This disclosure also provides a pooled IgG preparation containing a detectable amount of anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig). Those skilled in the art will see from this disclosure that a pooled IgG preparation containing a detectable amount of anti-FcεRI Ig is not provided by any of the methods of this disclosure. Therefore, this disclosure also provides a method for detecting anti-FcεRI Ig in a pooled IgG preparation.

[0108] This disclosure also provides methods for treating, preventing, and / or delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects, the methods comprising administering anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein. For example, this disclosure provides methods for treating conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects, the methods comprising administering anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein. This disclosure also provides methods for preventing conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects, the methods comprising administering anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein. This disclosure further provides a method for delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in a subject, the method comprising administering an anti-FcεRI Ig-reducing preparation, an anti-FcεRI Ig-reducing IgG preparation, or a pharmaceutical composition described herein.

[0109] This disclosure also provides anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein for use in treating, preventing, and / or delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein for use in treating conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein for use in preventing conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein for use in delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects.

[0110] This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein for treating, preventing, and / or delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein for treating conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein for preventing conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein, for delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects.

[0111] This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein in the manufacture of pharmaceuticals for the treatment, prevention, and / or delayed progression of immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein in the manufacture of pharmaceuticals for the treatment of immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or the pharmaceutical compositions described herein in the manufacture of pharmaceuticals for the prevention of immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects. This disclosure also provides the use of anti-FcεRI Ig-reducing preparations, anti-FcεRI Ig-reducing IgG preparations, or pharmaceutical compositions described herein in the manufacture of pharmaceuticals for delaying the progression of immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects.

[0112] This disclosure also provides a method for preparing an immunoglobulin (Ig) preparation from a plasma sample or fraction thereof, wherein the Ig preparation has a reduced adverse reaction in a subject, and the method comprises detecting the presence of anti-FcεRI Ig in a plasma sample or fraction thereof according to any method described herein, and producing an Ig preparation having a reduced adverse reaction.

[0113] In one example, the condition is selected from the group consisting of primary immunodeficiency, chronic inflammatory demyelinating polyneuropathy, and chronic immune thrombocytopenic purpura. In one example, the condition is primary immunodeficiency. In another example, the condition is chronic inflammatory demyelinating polyneuropathy. In yet another example, the condition is chronic immune thrombocytopenic purpura.

[0114] This disclosure also provides a method for preparing an immunoglobulin (Ig) preparation from a plasma sample or fraction thereof, wherein the Ig preparation has reduced adverse reactions in the subject, and the method comprises determining the rate of activation and / or degranulation of basophils and / or mast cells after exposure to the plasma sample or fraction thereof. (i) If the percentage of basophils expressing CD63 after exposure to the plasma sample or fraction 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, then include the plasma sample or fraction in the Ig preparation. (ii) If the percentage of basophils expressing CD63 after exposure to the plasma sample or fraction is greater than 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the total population of basophils exposed to the plasma sample or fraction, the plasma sample or fraction is excluded from the Ig preparation. (iii) If the percentage of basophils expressing CD203c after exposure to the plasma sample or fraction 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, then include the plasma sample or fraction in the Ig preparation. (iv) If the percentage of basophils expressing CD203c after exposure to the plasma sample or fraction is greater than 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the total population of basophils exposed to the plasma sample or fraction, the plasma sample or fraction is excluded from the Ig preparation. (v) If the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction 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, the plasma sample or fraction is included in the Ig preparation and / or (vi) If the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction is greater than 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the total population of mast cells exposed to the plasma sample or fraction, the plasma sample or fraction is excluded from the Ig preparation.

[0115] This disclosure also provides a method for identifying plasma donors suitable for the production of Ig preparations. [Brief explanation of the drawing]

[0116] [Figure 1-1] This is a graphical representation of the MRGPRX2 receptor-mediated activation profiles for cortistatin 14 (A, positive control), single plasma donors (B, C, including donor Z), affected products (D, F), and unaffected products (E, G). [Figure 1-2] Same as above. [Figure 2] This graph shows the activation of Hoxb8 mast cells by IVIG. Unaffected lots are shown as Ctrl-P. Affected lots are shown as AE-P (A) or hidden (B). The formulation buffer is used as a negative control (data not shown). [Figure 3] This graph shows the activation of Hoxb8 mast cells by SCIG. Unaffected lots are shown as Ctrl-H. Affected lots are shown as AE-H (A) or hidden (B). The formulation buffer is used as a negative control (data not shown). [Figure 4] This graph displays the mast cell activation profiles for 17 single-donor plasma samples (A, B) tested. Plasma sample 1 is from designated donor Z. Arrows indicate activation signals in plasma 1. [Figure 5] This graph displays the mast cell activation profiles for untreated plasma (A), IgG-depleted plasma (B), and IgE-depleted plasma (C) from donors 1, 2, and 3. Exclamation marks indicate suppression of Hoxb8 mast cell activation. [Figure 6] This graph displays the mast cell activation profile using the blockade of IgG binding to FcγRIIb by an Fc-based anti-CD16 / 32 antibody. The dotted line highlights the 5% threshold. [Figure 7] This graph displays the mast cell activation profile using soluble FcεRIα to block IgE binding to FcεRIα. Exclamation marks indicate partial suppression of Hoxb8 mast cell activation. The dotted line highlights the 5% threshold. [Figure 8] This graph displays the mast cell activation profile using cell-specific blockade by targeting BTK with ibrutinib. Exclamation marks indicate suppression of Hoxb8 mast cell activation. The dotted line highlights the 5% threshold. [Figure 9] This graph displays the mast cell activation profile using a C5aR antagonist to block C5a-dependent mast cell activation. The dotted line highlights the 5% threshold. [Figure 10-1] This graph shows the activation of basophil degranulation (CD63+) by IVIG using basophil donors 1(A), 2(B), and 3(C). Unaffected lots are shown as Ctrl-P. Affected lots are shown as AE-P or are hidden. The dotted line at the top highlights the 5% threshold, and the dotted line at the bottom highlights the baseline. [Figure 10-2] Same as above. [Figure 11-1] This graph shows the activation of basophil degranulation (CD63+) by SCIG using basophils from basophil donors 1(A), 2(B), and 3(C). Unaffected lots are shown as Ctrl-H. Affected lots are shown as AE-H. The dotted line at the top highlights the 5% threshold, and the dotted line at the bottom highlights the baseline. [Figure 11-2] Same as above. [Figure 12-1]This graph shows the activation of basophil degranulation (CD63+) in individual plasma samples using basophil donors 1(A), 2(B), and 3(C). Plasma sample 1 is from the designated donor Z. Arrows indicate the activation signal in plasma 1 in the undiluted state for all basophil donors. The upper dotted line highlights the 5% threshold, and the lower dotted line highlights the baseline. [Figure 12-2] Same as above. [Figure 13] This graph shows the activation of basophil degranulation, which is IL-3-deficient CD63+ (A) and CD203c+ (B), using individual plasma samples from donor 1. The dotted line highlights the 5% threshold. [Figure 14] This graph shows the activation of basophil degranulation in IL-3-deficient CD63+ (A) and CD203c+ (B) basophils using individual plasma samples from basophil donor 2. The dotted line highlights the 5% threshold. [Figure 15] This graph shows the activation of basophil degranulation, which is IL-3-deficient CD63+ (A) and CD203c+ (B), using individual plasma samples obtained from basophil donor 3. The dotted line highlights the 5% threshold. [Figure 16] This graph displays the activation of basophil degranulation (CD203c+) by SCIG(A) and IVIG(B) using basophil donor 1. The upper dotted line highlights the 5% threshold, and the lower dotted line highlights the baseline. Exclamation marks indicate activation signals exceeding the threshold. For each lot, basophil degranulation activation is plotted from left to right at SCIG(A) or IVIG(B) concentrations of 2, 20, 200, 2000, 20000, and 40000 μg / mL used in the assay. For each lot, on the right side of each panel, activation at a 2000 μg / mL SCIG(A) or IVIG(B) concentration spiked with anti-FcεRI is plotted as a stimulus control. [Figure 17]This graph displays the activation of basophil degranulation (CD203c+) by SCIG(A) and IVIG(B) using basophil donor 2. The upper dotted line highlights the 5% threshold, and the lower dotted line highlights the baseline. For each lot, the activation of basophil degranulation is plotted from left to right at SCIG(A) or IVIG(B) concentrations of 2, 20, 200, 2000, 20000, and 40000 μg / mL used in the assay. For each lot, on the right side of each panel, the activation at a 2000 μg / mL SCIG(A) or IVIG(B) concentration spiked with anti-FcεRI is plotted as a stimulus control. [Figure 18] This graph shows the activation of basophil degranulation (CD203c+) by SCIG(A) and IVIG(B) using basophil donor 3. The upper dotted line highlights the 5% threshold, and the lower dotted line highlights the baseline. For each lot, the activation of basophil degranulation is plotted from left to right at SCIG(A) or IVIG(B) concentrations of 2, 20, 200, 2000, 20000, and 40000 μg / mL used in the assay. For each lot, on the right side of each panel, the activation at a 2000 μg / mL SCIG(A) or IVIG(B) concentration spiked with anti-FcεRI is plotted as a stimulus control. [Figure 19-1] This graph displays basophil degranulation (CD203c+) from individual plasma samples using basophil donors 1(A), 2(B), and 3(C). The upper dotted line highlights the 5% threshold, the lower dotted line highlights the baseline, and spikes represent anti-FcεRI (stimulation control). For each plasma sample using basophil donors 1(A), 2(B), and 3(C), basophil degranulation is plotted from left to right at the following tested plasma dilutions: undiluted, 1:10, 1:100, 1:1000, 1:10000, and 1:100000. For each tested plasma sample, the 1:10 dilution spiked with anti-FcεRI is plotted on the right side of each panel as a stimulation control. [Figure 19-2] Same as above. [Figure 20-1]This graph displays the basophil degranulation profiles (CD203c+) in untreated plasma, IgG-depleted plasma, IgG-enriched plasma, and IgE-depleted plasma from donors 1 (A, D, G), 2 (B, E, H), and 3 (C, F, I). Plasma 1 samples are from the designated donor Z. Arrows indicate the suppression of activation signals in plasma 1 for all basophil donors for IgG-depleted plasma. The upper dotted line highlights the 5% threshold, the lower dotted line highlights the baseline, and spikes represent anti-FcεRI (stimulation control). [Figure 20-2] Same as above. [Figure 20-3] Same as above. [Modes for carrying out the invention]

[0117] overview Throughout this specification, unless otherwise specifically stated or the context requires, any reference to a single step, composition of a substance, group of steps, or group of compositions shall be construed as encompassing one or more (i.e., one or more) of those steps, compositions, groups of steps, or groups of compositions.

[0118] Those skilled in the art will understand that this disclosure is subject to variations and modifications other than those specifically described. It should be understood that this disclosure includes all such variations and modifications. This disclosure also includes, individually or collectively, all of the steps, features, compositions, and compounds referred to or indicated herein, as well as any and all combinations or any two or more of such steps or features.

[0119] This disclosure is intended for illustrative purposes only and should not be limited in scope by the specific examples described herein. Functionally equivalent products, compositions, and methods are clearly within the scope of this disclosure.

[0120] Any example in this disclosure shall apply mutatis mutandis to any other example in this disclosure unless otherwise specified.

[0121] Unless specifically defined otherwise, all technical and scientific terms used herein shall be construed to have the same meaning as those generally understood by those skilled in the art (e.g., immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0122] Unless otherwise indicated, the recombinant proteins, cell cultures, and immunological techniques used in this disclosure are standard procedures well known to those skilled in the art. Such techniques are described and explained through the following sources: J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984); J. Sambrook et al. Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989); TA Brown (ed.), Essential Molecular Biology: A Practical Approach, Vol. 1 and 2, IRL Press (1991); DMGlover and BDHames (eds.), DNA Cloning: A Practical Approach, Vol. 1-4, IRL Press (1995 and 1996); and FMAusubel et al. (eds.), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates to date); Ed Harlow and David Lane (eds.), Antibodies: A Laboratory Manual, Cold Spring Harbour Laboratory, (1988), and JEColigan et al. (eds.), Current Protocols in Immunology, John Wiley & Sons (including all updates to date).

[0123] The term "and / or," for example, "X and / or Y," is understood to mean either "X and Y" or "X or Y," and is construed as providing explicit support for both meanings or either meaning.

[0124] Throughout this specification, the word “comprise,” or variations such as “comprises” or “comprising,” will be understood to mean including the element, integer, or step, or group of elements, integers, or steps, that is described, but not to mean excluding any other element, element, or step, or group of elements, elements, or steps.

[0125] Selected definition The term "immunoglobulin (Ig)" should be understood as referring to proteins or antibodies present in serum and immune system cells. There are several types of Ig, such as IgG, IgE, IgM, IgD, IgA, and IgY.

[0126] As used herein, “anti-Fc-epsilon receptor I (FcεRI) immunoglobulin (Ig)” or “anti-FcεRI Ig” should be understood to refer to FcεRI, its fragments, or any Ig that binds to the epitopes of FcεRI.

[0127] As used herein, the term “FcεRI-mediated signaling pathway” refers to the crosslinking of FcεRIs that results in the activation and / or degranulation of basophils and / or mast cells.

[0128] The term "plasma" refers to the straw-colored / pale-yellow component of blood obtained from one or more blood donors. Methods for obtaining plasma from donors are obvious to those skilled in the art and / or will be described herein. For example, plasma is obtained by removing red blood cells from donated blood. For example, plasma is obtained by plasmapheresis.

[0129] The terms “plasma sample” or “fraction thereof” refer to plasma that has been fractionated to isolate one or more desirable protein components from the plasma. For example, plasma may be fractionated to isolate cold precipitates (proteins that precipitate from solution when units of fresh frozen plasma are slowly thawed in cold conditions) and cold supernatants (also known as decryoplasma). For example, plasma may be fractionated by ethanol precipitation, as described in U.S. Patent No. 3,301,842, to yield IgG-containing Onclay fraction, Cohn fraction, ammonium sulfate precipitate, or precipitate A (KNA) from the plasma. The plasma fraction includes II+III precipitates produced according to the Cohn method, 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 I+II+III precipitates produced according to Method 10, Cohn et al. J. Am; Chem. Soc., 72, 465-474 (1950), and the method of Deutsch et al. J. Biol. Chem. 164, 109-118 (1946), or precipitate A of Nitschmann and Kistler Vox Sang. 7, 414-424 (1962), Helv. Chim. Acta 37, 866-873 (1954). For example, plasma can be fractionated by octanoic acid fractionation, as described in European Patent Application No. 893450. Typically, the Kohn fraction, Kistler / Nischemann precipitate A (KN A), exists as a suspension paste. Other purification techniques, including chromatography, may be used.

[0130] As used herein, “IgG intermediate product” refers to any material obtained between manufacturing steps, for example, the supernatant or fraction of the starting material.

[0131] As used herein, “mast cells” should be understood to refer to granular tissue commensal cells known for their host cell responses, allergic responses, and vascular homeostasis.

[0132] As used herein, “basophils” should be understood to refer to the largest type of granulocyte, which accounts for approximately 0.5%–1% of circulating white blood cells. Their activation and degranulation are involved in inflammatory responses during immune responses, as well as in the formation of acute and chronic allergic diseases.

[0133] The term "protein" should be understood to include a single polypeptide chain, i.e., a series of consecutive amino acids linked by peptide bonds, or a series of polypeptide chains covalently or non-covalently bonded to one another (i.e., a polypeptide complex). For example, a series of polypeptide chains can be covalently bonded using suitable chemical bonds or disulfide bonds. Examples of non-covalent bonds include hydrogen bonds, ionic bonds, van der Waals forces, and hydrophobic interactions.

[0134] The terms "polypeptide" or "polypeptide chain" will be understood from the preceding paragraph to mean a series of consecutive amino acids linked by peptide bonds.

[0135] Those skilled in the art will know that an "antibody" generally consists of multiple polypeptide chains, for example, a light chain variable region (V L ) a polypeptide containing a heavy chain variable region (V H It will be recognized that antibodies are thought to be proteins containing a polypeptide comprising a variable region consisting of ). Antibodies also generally contain constant domains, some of which, in the case of heavy chains, can be located within a constant region containing a constant fragment or crystallizable fragment (Fc). H and V LThese interact to form an Fv containing an antigen-binding region that can specifically bind to one or more closely related antigens. Generally, the light chain from mammals is either a κ light chain or a λ light chain, and the heavy chain from mammals is α, δ, ε, γ, or μ. Antibodies can be any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), a class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or a subclass. The term “antibody” also encompasses humanized antibodies, primate-derived antibodies, human antibodies, syn-humanized antibodies, and chimeric antibodies.

[0136] As used herein, “variable region” refers to a portion of the light and / or heavy chain of an antibody as defined herein, which can specifically bind to an antigen and includes the amino acid sequences of complementarity-determining regions (CDRs), i.e., CDR1, CDR2, and CDR3, as well as framework regions (FRs). An example variable region includes three or four FRs (e.g., FR1, FR2, FR3, and optionally FR4) along with three CDRs. H This refers to the variable region of the heavy chain. L This refers to the variable region of the light chain.

[0137] As used herein, the terms “detect” or “to detect” refer to the identification of the presence or existence of anti-FcεRI Ig in a plasma sample.

[0138] As used herein, the terms “contact” or “to come into contact” are used to refer to direct or indirect interactions or associations between a ligand or reagent described herein and a molecule. For example, a ligand (such as FcεRI or a fragment or epitope thereof) directly or indirectly binds to anti-FcεRI Ig in a plasma sample or fraction thereof. The terms also encompass interactions, for example, between a first protein (such as a detection protein) and, for example, a second molecule (such as an antibody).

[0139] As used herein, the term “binding” in relation to protein-antigen interactions means that the interaction depends on the presence of a specific structure on the antigen (e.g., an antigenic determinant or epitope). For example, antibodies generally recognize and bind to specific protein structures, not proteins themselves. If an antibody binds to epitope “A”, in a reaction involving labeled “A” and the antibody, the presence of molecules containing epitope “A” (or free, unlabeled “A”) will reduce the amount of labeled “A” that binds to the antibody.

[0140] Where used herein, the terms “particularly bind” or “specifically bind” should be understood to mean that the antibodies of this disclosure react to or associate with a particular antigen (e.g., S protein or RBD) more frequently, more rapidly, for a longer duration, and / or with higher affinity than surrogate antigens. For example, an antibody binds to an S protein or RBD with significantly higher affinity (e.g., 20-fold or 40-fold or 60-fold or 80-fold to 100-fold or 150-fold or 200-fold higher affinity) binding activity, more easily, and / or for a longer duration than an antibody binds to an antigen commonly recognized by other antigens, for example, by a multireactive native antibody (i.e., a naturally occurring antibody known to bind to a variety of naturally occurring antigens found in humans). Generally, though not always, references to binding should be understood to mean specific binding, and each term should be understood to provide explicit support for the other terms.

[0141] As used herein, the term “epitope” (synonym “antigenic determinant”) should be understood to mean a region of a protein to which a protein containing an antibody variable region binds. The term is not necessarily limited to the specific residue or structure to which the protein contacts. For example, the term includes a region spanning amino acids contacted by a protein, and / or 5 to 10, 2 to 5, or 1 to 3 amino acids outside this region. In some examples, an epitope includes a series of discontinuous amino acids that are positioned close to each other when the protein is folded, i.e., a “structural epitope.” Those skilled in the art will also recognize that the term “epitope” is not limited to peptides or polypeptides. For example, the term “epitope” includes a chemically active surface classification of a molecule, such as a sugar side chain, phosphoryl side chain, or sulfonyl side chain, which in certain examples may have specific three-dimensional structural properties and / or specific charge properties.

[0142] The term "affinity chromatography resin" should be understood, as will be apparent to those skilled in the art and / or as described herein, to mean a resin containing an affinity chromatography ligand (e.g., FcεRI, or a fragment or epitope thereof) bound to a matrix.

[0143] The term "ligand" should be understood as a molecule immobilized on the matrix of a chromatography column that interacts with anti-FcεRI Ig. For example, a ligand may include FcεRI or a fragment or epitope thereof. In one example, the ligand is FcεRI or a fragment or epitope thereof. In another example, the ligand is an FcεRI antibody or a fragment thereof. In yet another example, the ligand is an FcεRI DNA aptamer.

[0144] The term “matrix” should be understood to mean a support to which molecules can be directly or indirectly bound. The matrix may include any substrate material that can provide a physical support for the compositions described herein. The material may be a naturally occurring material, a synthetic material, or a modification of a naturally occurring material. Suitable matrix materials include glass fibers, polyester, cellulose, rayon, silicon, silicon wafer chips, graphite, mirrors, laminates, films, ceramics, plastics (including polymers such as poly(vinyl chloride), cycloolefin copolymers, agarose gels or beads, polyacrylamide, polyacrylate, polyethylene, polypropylene, poly(4-methylbutene), polystyrene, polymethacrylate, poly(ethylene terephthalate), polytetrafluoroethylene (PTFE or Teflon®), nylon, and poly(vinyl butyrate)), germanium, gallium arsenide, gold, silver, Langmuir-Bludget films, flow-through chips, etc., which may be used either by themselves or in combination with other materials. Additional rigid materials such as glass, which may contain silica and further include glass that can be used as bioglass, for example, may be considered. Other materials that may be used include porous materials such as controlled-pore glass beads, cross-linked beads Sepharose® or agarose resin, or cross-linked bis-acrylamide and azalactone copolymers.

[0145] The term “pharmaceutical composition” should be understood to mean a formulation of Ig having compounds generally accepted in the art for the delivery of Ig to mammals. Exemplary compounds include all pharmaceutically acceptable carriers, diluents, or excipients thereof.

[0146] The terms “to treat,” “to cure,” or “to treat” should be understood to mean administering a therapeutically effective dose of Ig such that one or more symptoms or characteristics of the condition are reduced in the subject, or the subject is no longer clinically diagnosed as having the condition.

[0147] The terms “prevention,” “prevent,” or “prevention” include providing preventive measures regarding the occurrence or recurrence of a particular condition in a subject. The subject is predisposed to or at risk of developing the condition but has not yet been diagnosed with the condition.

[0148] As used herein, the phrase “delay the progression of” includes reducing or delaying the progression of a condition and / or two or more symptoms of a condition in the subject.

[0149] The term "condition" should be understood to mean the presence or health state of an individual requiring treatment with Ig.

[0150] The term "subject" should be understood to mean any animal, including humans and, for example, mammals. Examples of subjects include, but are not limited to, humans and non-human primates. For example, the subject is a human.

[0151] Fc epsilon receptor I (FcεRI) This disclosure provides a method for identifying anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof.

[0152] As used herein, the term “Fc epsilon receptor I (FcεRI)” should be understood to refer to antigens present on mast cells, basophils, epidermal Langerhans cells, eosinophils, and other antigen-presenting cells. FcεRIs are members of the multimeric receptor and associated antigen / Fc receptor family, sharing conserved structural features and similar roles in initiating intracellular signaling cascades. In humans, two types of FcεRIs are expressed on the cell surface: a tetrameric receptor (αβγ2) consisting of an α chain, a β chain, and a homodimeric γ chain, and a trimer receptor (αγ2) consisting of an α chain and a homodimeric γ chain.

[0153] FcεRIs are involved in the activation and degranulation of mast cells and basophils. Antigen crosslinking of FcεRIs, for example via IgE and / or IgG binding, initiates multiple signaling pathways known as the "FcεRI-mediated signaling pathway," which regulate diverse effector responses. These include the secretion of allergy mediators and the induction of cytokine gene transcription, leading to the secretion of molecules such as interleukin-4, interleukin-6, tumor necrosis factor-α, and granulocyte-macrophage colony-stimulating factor. Thus, FcεRIs are central to the induction and maintenance of allergic responses.

[0154] To detect, identify, and screen for anti-FcεRI Ig This disclosure provides a method for detecting anti-FcεRI Ig in a plasma sample or fraction thereof. In addition, this disclosure provides a method for screening for anti-FcεRI Ig in a plasma sample or fraction thereof.

[0155] This disclosure also provides a method for identifying plasma samples or fractions suitable for administration to a subject. This disclosure also provides a method for identifying plasma samples or fractions for use in preparing immunoglobulin (Ig) preparations for administration to a subject. Furthermore, this disclosure provides a method for screening plasma samples or fractions to determine their suitability for administration to a subject. This disclosure also provides a method for screening plasma samples or fractions for use in preparing immunoglobulin (Ig) preparations for administration to a subject.

[0156] Those skilled in the art will see that plasma samples or fractions containing detectable amounts of anti-FcεRI Ig are unsuitable for administration to subjects and / or for use in preparing Ig preparations for administration to subjects.

[0157] As used herein, “detectable amount” should be understood to mean any amount or quantity of anti-FcεRI Ig that induces activation of the FcεRI-mediated signaling pathway. Those skilled in the art will see from the disclosure herein that while anti-FcεRI Ig may still be present in plasma samples or fractions thereof, the amount is insufficient to induce activation of the FcεRI-mediated signaling pathway. For example, activation of the FcεRI-mediated signaling pathway can occur when the sample contains more than 40 μg / mL of anti-FcεRI IgG and / or more than 0.1 kU / L of anti-FcεRI IgE.

[0158] For example, activation of the FcεRI-mediated signaling pathway can occur when, after exposure to the sample, the proportion of basophils expressing CD63 exceeds 5% of the total population of basophils exposed to the sample, and / or the proportion of basophils expressing CD203c exceeds 5% of the total population of basophils exposed to the sample, and / or the proportion of mast cells expressing CD107a exceeds 5% of the total population of mast cells exposed to the sample. For example, the proportion of basophils expressing CD63 and / or CD203c is relative to the total proportion of basophils exposed to the plasma sample or fraction. For example, the proportion of mast cells expressing CD107a is relative to the total proportion of mast cells exposed to the plasma sample or fraction.

[0159] In an alternative example, a plasma sample or fraction that does not contain a detectable amount of anti-FcεRI Ig is suitable for administration to a subject. For example, the method further includes administering a plasma sample to a subject if anti-FcεRI Ig is not present or detectable in the plasma sample or fraction.

[0160] This disclosure further provides a method for identifying suitable subjects for plasma donation for the manufacture of Ig preparations. In addition, this disclosure provides a method for screening subjects for plasma donation for the manufacture of Ig preparations. This disclosure also provides a method for identifying suitable plasma donations for the manufacture of Ig preparations.

[0161] Those skilled in the art will see that a plasma sample or fraction from a subject exhibiting a detectable amount of anti-FcεRI Ig indicates that the subject is not a suitable candidate for plasma donation for the production of Ig preparations. Furthermore, a plasma sample or fraction exhibiting a detectable amount of anti-FcεRI Ig indicates that the plasma sample or fraction is not suitable for the production of Ig preparations.

[0162] In an alternative example, a plasma sample or fraction from a subject that does not contain a detectable amount of anti-FcεRI Ig indicates that the subject is a suitable candidate for plasma donation for the production of an Ig preparation. Furthermore, a plasma sample or fraction that does not exhibit a detectable amount of anti-FcεRI Ig indicates that the plasma sample or fraction is suitable for the production of an Ig preparation. For example, the method further includes donating a plasma sample or fraction for the production of an Ig preparation if no detectable amount of anti-FcεRI Ig is present.

[0163] Methods for such detection, screening, and / or identification will be apparent to those skilled in the art and / or are described herein. Exemplary methods for detecting, screening, and / or identifying anti-FcεRI Ig in plasma samples or fractions thereof or subjects include lateral flow assays and ELISAs using ligands that bind to anti-Fc epsilon receptor I (FcεRI) Ig (e.g., FcεRI or its fragments or epitopes). Methods for determining the level of anti-FcεRI Ig will be apparent to those skilled in the art and / or are described herein.

[0164] In one example, the method includes performing an enzyme-linked immunosorbent assay (ELISA). In another example, the method includes performing a fluorescence-linked immunosorbent assay (FLISA). In yet another example, the method includes performing a lateral flow immunoassay.

[0165] Enzyme-linked immunosorbent assay (ELISA) and fluorescence-linked immunosorbent assay (FLISA) This disclosure provides an ELISA or FLISA for detecting, identifying, and / or screening anti-FcεRI Ig in a plasma sample or fraction thereof, wherein the assay comprises a ligand (e.g., FcεRI or a fragment or epitope thereof).

[0166] Standard solid-phase ELISA or FLISA formats are particularly useful for determining the concentration of proteins (e.g., antibodies or Ig) from various samples. In one form, such assays involve immobilizing the biological sample on a solid matrix.

[0167] A ligand (e.g., FcεRI or a fragment or epitope) that specifically binds to a protein (i.e., anti-FcεRI Ig) in the plasma sample of that fraction is brought into direct contact with the immobilized ligand to form a direct bond with any of its target proteins present in the sample. This ligand is generally labeled with a detectable reporter molecule, for example, fluorescently labeled (e.g., FITC or Texas Red) or fluorescent semiconductor nanocrystals (as described in US6,306,610) in the case of FLISA, or with an enzyme (e.g., horseradish peroxidase (HRP), alkaline phosphatase (AP), or β-galactosidase) in the case of ELISA, or alternatively, a labeled protein or antibody that binds to the first antibody can be used. After washing to remove any unbound proteins, the label is detected either directly in the case of fluorescent labeling, or, in the case of enzymatic labeling, by adding a substrate such as hydrogen peroxide, TMB, or toluidine, or 5-bromo-4-chloro-3-indole-beta-D-galaotopyranoside (x-gal).

[0168] Such ELISA or FLISA-based systems are suitable for quantifying the amount of protein in a sample by calibrating the detection system against a known standard amount to which the ligand binds, such as anti-FcεRI Ig.

[0169] Those skilled in the art will understand that the methods of this disclosure are suitable for use in capture enzyme-linked immunosorbent assays (ELISA) or enzyme-mediated immunoassays (EIA). As used herein, the terms “ELISA,” “sandwich ELISA,” “capture ELISA,” or “EIA” refer to immobilizing a ligand (specific to anti-FcεRI Ig) on ​​a matrix such as a membrane, polystyrene or polycarbonate microwells, polystyrene or polycarbonate dipsticks, or a glass support, followed by the addition of a certain amount of plasma sample. The molecule is then “bound” or “captured.” The captured anti-FcεRI Ig can be detected by a detection protein or detection antibody that can covalently bind to the enzyme, or it can be detected itself by the addition of a secondary protein or antibody bound to the enzyme.

[0170] Those skilled in the art will see that the assay formats described herein can conform to, for example, automated screening processes or high-throughput formats such as microarray formats as described in Mendoza et al., 1999. Furthermore, variations of the above assays will be apparent to those skilled in the art, such as competitive ELISA.

[0171] Lateral flow assay This disclosure provides a lateral flow assay for detecting and / or screening anti-FcεRI Ig in plasma samples or fractions thereof, the assay comprising a ligand and a lateral flow assay device.

[0172] Lateral flow assays, also known as "immunochromatography strip tests," operate on the same principle as enzyme-linked immunosorbent assays (ELISA). Essentially, these tests involve running a liquid sample along the surface of a membrane or filter paper containing reactive molecules that produce a visually positive or negative result depending on the presence of a specific analyte (e.g., anti-FcεRI Ig).

[0173] A lateral flow assay device is a test system for detecting a target analyte or component in a sample, for example, detecting the presence of anti-FcεRI Ig in a plasma sample or a fraction thereof. A lateral flow assay device is a device configured to receive a sample in a sample region and to provide lateral movement of the sample from the sample region to the detection region by capillary action, for example, via wicking. In certain examples, the lateral flow assay device further includes one or more conjugation regions, and the lateral flow assay device is configured to provide a lateral flow of the sample from the sample region to one or more conjugation regions before reaching the detection region. In a relevant example of a lateral flow assay device, the sample region is in contact with a conjugation region, and the conjugation region is in contact with one end of the detection region, such that the lateral flow assay device is configured to allow the sample to flow from the sample region to the conjugation region and finally to the detection region. In certain examples of lateral flow assay devices, the device further includes an absorption region in contact with the detection region, such that the device is configured to allow the flow of sample from the sample region to the detection region and finally to the absorption region.

[0174] Lateral assay devices typically have a matrix to which optional sample areas, optional conjugation areas, detection areas, and optional absorption areas are mounted. The matrix ("backing card") provides support for the pad and membrane of the actual assay and, in other respects, does not participate in the reaction or flow of the sample and analyte. The backing card is made from, for example, polyvinyl chloride (PVC). The assembly of the pad and membrane on the backing card is typically housed in a plastic housing, although this is not required. The housing may have two or more openings ("sample ports") on the sample pad for sample application. The control zone and test zone are visible (e.g., through the openings or windows) for detecting or measuring the bound label. The housing prevents the user from applying the sample to a location other than the sample pad. The housing also helps protect the strip from unintended splashes onto the membrane. External markings on the housing may also be used to indicate the location of the test and control lines and to provide other information. The housing can be obtained as a ready-made cassette or custom-designed to fit around the strip. Internal pins and bars can be used to hold the strip in place relative to the sample port and visual window. They hold the material in fluid communication with each other while the test strip is running.

[0175] The "sample area," if present, receives the sample upon application and facilitates the uniform distribution of the sample into the detection area or conjugation area (if present). It also influences the rate at which the liquid enters the detection area and can prevent overflow of the device. In addition, the sample pad may also contain additional components such as proteins, detergents, viscosity enhancers, and buffer salts to process the sample (e.g., in the case of blood samples, this may be for separating sample components, removing interference, adjusting pH, increasing viscosity, solubilizing components, and / or preventing nonspecific binding between the conjugate and the analyte or other components, or with the reaction membrane).

[0176] The “conjugate region” or “conjugation region,” if present, includes a dry, movable composition containing a labeled antibody or labeled protein. When the sample flows into the conjugation region, the labeled antibody or labeled protein detaches from the conjugate region material and moves forward into the detection region along with the sample. Where applicable, the conjugation region also includes a dry, movable control conjugate.

[0177] In other examples, the lateral flow assay device does not include a separate conjugation region. In such examples, the sample is mixed in a separate container with a composition containing a labeled antibody or labeled protein disclosed herein before moving along the lateral flow assay device. Such a device may be called a lateral flow assay dipstick. For example, a plasma sample or fraction thereof from a subject may be brought into contact with a composition described herein in a separate container to produce a mixed solution, and then a lateral flow assay device including a detection region may be immersed in the solution so that it moves along the detection region to the test zone and the control zone.

[0178] The "detection region" is typically a membrane containing a test zone and a control zone, both containing irreversibly bound capture reagents, such as an antibody or protein, an antibody against a labeled protein or labeled antibody, or a capture reagent such as streptavidin. Typically, the reaction membrane is made from a polymer such as nitrocellulose, polyvinylidene fluoride, nylon, or polyethersulfone. Nitrocellulose is an exemplary choice for the reaction membrane. Nitrocellulose membranes electrostatically bind to proteins (such as antibodies or biotin-binding proteins) through the interaction of the strong dipoles of nitrate esters with the strong dipoles of peptide bonds within the protein.

[0179] Lateral flow assay devices may also include an "adsorption region" or "absorption region." The adsorption region is located at the distal end of the detection region and holds the remaining sample. It draws liquid through the membrane and collects the processed liquid. Furthermore, it increases the total volume of sample that can enter the detection region.

[0180] Suitable materials for the sample area, conjugation area, or detection area that may be included in the lateral flow assay device described herein include, but are not limited to, organic or inorganic polymers, as well as natural and synthetic polymers including glass fibers, cellulose, nylon, cross-linked dextran, various chromatographic papers, and nitrocellulose. It will be understood that suitable materials allow the sample to flow laterally along the device described herein via capillary action. In certain examples, the detection area is a nitrocellulose membrane. In certain examples, the sample area and conjugation area may be composed of the same material. In certain examples, the lateral flow assay device includes a detection area and a sample area in capillary contact. Suitable commercially available materials will be known to those skilled in the art. Commercially available materials may be used for the sample area, conjugation area, and / or detection area that may constitute the lateral assay device described herein.

[0181] The lateral flow assay device may further include a sample filter membrane applied to the sample area. The sample filter membrane may consist of any suitable material, but is not limited to, a hydrophobic material capable of filtering cells (e.g., blood cells) from a fluid. Suitable sample membranes should be obvious to those skilled in the art and may have filter sizes ranging from about 0.22 μm to about 10 μm, for example.

[0182] In one example, a sample is applied to the sample area of ​​a lateral flow assay device, and the device is then incubated. Incubation includes allowing the device to be maintained at a temperature, e.g., room temperature (e.g., approximately 20°C to approximately 25°C), so that the sample flows from the sample area to the detection area. In an example further including a conjugation area, incubation includes allowing the lateral flow device to be maintained at a temperature, e.g., room temperature (e.g., approximately 20°C to approximately 25°C), so that the sample flows from the sample area to the conjugation area and then to the detection area.

[0183] In one example, the lateral flow assay device is incubated after applying the sample to the sample area for approximately 2 to 20 minutes, 2 to 15 minutes, or 2 to 10 minutes. For example, the lateral flow assay device is incubated for approximately 10 to 15 minutes after applying the sample to the sample area.

[0184] In one example, the lateral flow assay device may further include a control component immobilized in a control zone of the detection area. In one example, the detection area of ​​the lateral flow assay device is configured such that the sample passes through a test zone preceding the control zone. In one example, the assay may further include inspection of the signal on the control line to confirm the effective operation of the lateral flow assay device. The inspection may include visual confirmation of the signal on the control line.

[0185] In one example, evaluation may include quantitative measurement of molecules trapped in the test zone and / or control zone. In another example, evaluation may include semi-quantitative or qualitative evaluation of the test zone, e.g., detection of signals exceeding a predetermined threshold. The preferred means of evaluating the test zone will depend on the signals generated by the test zone. For example, detection may be optical, thermal, magnetic, or electrochemical. In another example, evaluation may include quantitative or qualitative measurement of signals from, for example, fluorescent dyes or colloidal metals. Evaluation may be performed visually. Evaluation may be performed using a smartphone. In certain examples, evaluation may include the use of a portable fluorometer. Commercially available devices for measuring signals from lateral flow assay devices will be familiar to those skilled in the art.

[0186] Detectable signs As used herein, “detectable label” is a marker that produces or can induce the production of a product that can be detected by a molecular or atomic tag, or by an optical or other signal, or by visually or by using a suitable detector. Detectable labels are well known in the art and include, for example, radioactive labels, enzymes, fluorescent labels, luminescent labels, bioluminescent labels, magnetic labels, prosthetic groups, contrast agents, and sonicants.

[0187] Commonly used fluorescent labels include, but are not limited to, cyanines and indocyanines such as Alexa, Cy5, and Cy5.5, as well as fluorescein isothiocyanate (FITC). Similarly, but are not limited to, fluorescent labels useful for implementing this disclosure, including, 1,5-IAEDANS; 1,8-ANS; 4-methylumbelliferone; 5-carboxy-2,7-dichlorofluorescein; 5-carboxyfluorescein (5-FAM); 5-carboxynaptofluorescein (pH10); 5-carboxytetramethylrhodamine (5-TAMRA); 5-FAM (5-carboxyfluorescein); 5-HAT (hydroxytryptamine); 5-hydroxytryptamine (HAT); 5-ROX (carboxy-X-rhodamine); 5-TAMRA (5-carboxytetramethylrhodamine); 6-carboxyrhodamine 6C; 6-CR 6G; 6-JOE; 7-amino-4-methylcoumarin; 7-aminoactinomycin D (7-AAD); 7-hydroxy-4-methylcoumarin; 9-amino-6-chloro-2-methoxyacrycidine; ABQ; acid fuchsin; ACMA (9-amino-6-chloro-2-methoxyacrycidine); acridine orange + DNA; acridine orange + RNA; acridine orange, both DNA and RNA; acridine red; acridine yellow; acrylflavin; acrylflavin foilgen SITSA; aequorin (photoprotein); Alexa Fluor350; Alexa Fluor430; Alexa Fluor488; Alexa Fluor532; Alexa Fluor546; Alexa Fluor568; Alexa Fluor594; Alexa Fluor633; Alexa Fluor647; Alexa Fluor660; Alexa Fluor680; Alizarin complexone; Alizarin red; Allophycocyanin (APC); AMC, AMCA-S; AMCA (aminomethylcoumarin); AMCA-X; Aminoactinomycin D; Aminocoumarin; Aminomethylcoumarin (AMCA); Aniline blue; Anthrosyl stearate; APC (allophycocyanin); APC-Cy7; APTRA-BTC = specific pigment, Zn 2+;APTS; Astrazon Brilliant Red 4G; Astrazon Orange R; Astrazon Red 6B; Astrazon Yellow 7GLL; Atabrine; ATTO-TAG CBQCA; ATTO-TAG FQ; Auramine; Aurphosphine G; Aurphosphine; BAO9 (Bisaminophenyl Oxadiazole); BCECF (High pH); BCECF (Low pH); Berberine Sulfate; Beta-Lactamase; BFP Blue Shift GFP (Y66H); Blue Fluorescent Protein; BFP / GFP FRET Bimane; Bisbenzimide; Bisbenzimide (Hoechst); bis-BTC = Ratio Dye, Zn 2+ ; Blancophor FFG; Blancophor SV; BOBO-1; BOBO-3; Bodipy 492 / 515; Bodipy 493 / 503; Bodipy 500 / 510; Bodipy 505 / 515; Bodipy 530 / 550; Bodipy 542 / 563; Bodipy 558 / 568; Bodipy 564 / 570; Bodipy 576 / 589; Bodipy 581 / 591; Bodipy 630 / 650-X; Bodipy 650 / 665-X; Bodipy 665 / 676; Bodipy Fl; Bodipy FL ATP; Bodipy Fl-Ceramide; Bodipy R6G SE; Bodipy TMR; Bodipy TMR-X Conjugate; Bodipy TMR-X,SE; Bodipy TR; Bodipy TR ATP; Bodipy TR-X SE; BO-PRO-1; BO-PRO-3; Brilliant Sulfoflavin FF; BTC - Ratio Dye Ca 2+ ; BTC-5N-atio Dye, Zn 2+ ; Calcein; Calcein Blue; Calcium Crimson; Calcium Green; Calcium Green-1 Ca 2+ Dye; Calcium Green-2 Ca 2+ ; Calcium Green-5N Ca 2+ ; Calcium Green-C18 Ca 2+Calcium Orange; Chalcoflor White; Carboxy-X-Rhodamine (5-ROX); Cascade Blue; Cascade Yellow 399; Catecholamine; CCF2 (GeneBlazer); CFDA; CFP-Cyanide Fluorescent Protein; CFP / YFP; FRET; Chlorophyll; Chromomycin A; Chromomycin A; CL-NERF (Colorant; pH); CMFDA; Coelenterazine; Coelenterazine cp(Ca 2+ Pigments); coelenterazine f; coelenterazine fcp; coelenterazine h; coelenterazine hcp; coelenterazine ip; coelenterazine n; coelenterazine O; coumarin phalloidin; C-phycocyanin; CPM methylcoumarin; CTC; CTC formazan; Cy2; Cy3.1 8; Cy3.5; Cy3; Cy5.1 8;Cy5.5;Cy5;Cy7;Cyanide GFP;Cyclic AMP Fluorosensor (FiCRhR);CyQuant Cell Proliferation Assay;Dabsil;Dansil;Dansilamine;Dansilcadaverine;Dansilchloride;DansilDHPE;Dansilfluoride;DAPI;Dapoxyl;Dapoxyl2;Dapoxyl3;DCFDA;DCFH (Dichlorodihydrofluorescein diacetate);DDAO;DHR (Dihydrorhodamine 123);Di-4-ANEPPS;Di-8-ANEPPS (Non-comparable);DiA(4-di-16-ASP);Dichlorodihydrofluorescein diacetate (DCFH) ;DiD-lipophilic tracer;DiD(DiIC18(5));DIDS;Dihydrorhodamine 123(DHR);DiI(DiIC18(3));Dinitrophenol;DiO(DiOC18(3));DiR;DiR(DiIC18(7));DM-NERF(high pH);DNP;Dopamine;DsRed;Red fluorescent protein;DTAF;DY-630-NHS;DY-635-NHS;EBFP;ECFP;EGFP;ELF97;Eosin;Erythrosine;Erythrosine ITC;Ethidium bromide;Ethidium homodimer-1(EthD-1);Eucrysin;Eucolite; Europium(III) chloride; EYFP; Fast Blue; FDA; Foilgen (pararoseaniline); FIF (formaldehyde-induced fluorescence); FITC; FITC antibody; Furazo Orange; Fluo-3; Fluo-4; Fluorescein (FITC); Fluorescein diacetic acid; Fluoroemerald; Fluorogold (hydroxystilvamidine); Fluororuby; FluorX; FM1-43; FM4-46; Fura Red (high pH); Fura Red / Fluo-3; Fura-2, high calcium; Fura-2, low calcium; Fura-2 / BCECF; Genacryl Brilliant Red B; Genacryl Brilliant Yellow 10GF; Genacryl Pink 3G; Genacryl Yellow 5GF; Gene Brother (CCF2); GFP (S65T); GFP Redshift (rsGFP), GFP wild-type, non-UV excited (wtGFP); GFP wild-type, UV excited (wtGFP); GFPuv; Gloxalic acid; Granular blue; Hematoporphyrin; Hoechst 33258; Hoechst 33342; Hoechst 34580; HPTS; Hydroxycoumarin; Hydroxystilbamidine (Fluorogold); Hydroxytryptamine; Indo-1, high calcium; Indo-1, low calcium; Indodicarbocyanin (DiD); Indotricarbocyanin (DiR); Intrawhite Cf; JC-1; JO-JO-1; JO-PRO- 1; LaserPro; Laurodane; LDS751 (DNA); LDS751 (RNA); Lowcophore PAF; Lowcophore SF; Lowcophore WS; Lysamin Rhodamine; Lysamin Rhodamine B; LIVE / DEAD Kit Animal Cells, Calcein / Ethidium Homodimer; LOLO-1; LO-PRO-1; Lucifer Yellow; Lithotracker Blue; Lithotracker Blue White; Lithotracker Green; Lithotracker Red; Lithotracker Yellow; Lysosensor Blue, Lithosensor Green; Lithosensor Yellow / Blue; Mag Green; Magdalena Red (Phloxine B); Mag-Fura Red; Mag-Fura-2; Mag-Fura-5; Mag-Indo-1; Magnesium Green; Magnesium Orange; Malachite Green; Marina Blue; Maxilon Brilliant Flavin 10GFF; Maxilon Brilliant Flavin 8GFF;Merocyanine; Methoxycoumarin; Mitotracker Green FM; Mitotracker Orange; Mitotracker Red; Mitramycin; Monobromoviman; Monobromoviman (mBBr-GSH); Monobromoviman; MPS (Methylgreen pyronin stilbene); NBD; NBDamine; Nile Red; Nitrobenzoxazidol; Noradrenaline; Nuclear Fast Red; Nuclear Yellow; Nylosanc Brilliant Iavin E8G; Oregon Green; Oregon Green 488-X; Oregon Green; Oregon Green 488; Oregon Green 500; Oregon Green 514; Pacific Blue; Pararoseaniline (Feulgen); PBFI; PE-Cy5; PE-Cy7; PerCP; PerCP-Cy5.5; PE-Texas Red [Red 613]; Phloxine B (Magdalene Red); Holwhite AR; Holwhite BKL; Holwhite Rev ;Holwait RPA;Phosphine 3R;Photoresist;Phycoerythrin B[PE];Phycoerythrin R[PE];PKH26(Sigma);PKH67;PMIA;Pontochrome Blue Black;POPO-1;POPO-3;PO-PRO-1;PO-PRO-3;Primulin;Procyon Yellow;Propidium Iodide (PI);PyMPO;Pyrene;Pyronine;Pyronine B;Pyrozal Brilliant Flavin 7GF;QSY7;Ki Naklin Mustard; Red 613 [PE-Texas Red]; Resolfin; RH414; Rhod-2; Rhodamine; Rhodamine 110; Rhodamine 123; Rhodamine 5GLD; Rhodamine 6G; Rhodamine B; Rhodamine B200; Rhodamine B Extra; Rhodamine BB; Rhodamine BG; Rhodamine Green; Rhodamine Faricidine; Rhodamine Phalloidin; Rhodamine Red; Rhodamine WT; Rose Bengal; R-F Phycocyanin; R-phycoerythrin (PE); rsGFP; S65A; S65C; S65L; S65T; SapphireGFP; SBFI; Serotonin; Sebron Brilliant Red 2B; Sebron Brilliant Red 4G; Sebron Brilliant Red B; Sebron Orange; Sebron Yellow L; sgBFP; sgBFP (Super Glow BFP); sgGFP; sgGFP (Super Glow GFP); SITS; SITS (Primulin);SITS (Stilbene Isothiosulfonic Acid); SNAFL Calcein; SNAFL-1; SNAFL-2; SNARF Calcein; SNARF1; Sodium Green; Spectrum Aqua; Spectrum Green; Spectrum Orange; Spectrum Red; SPQ (6-Methoxy-N-(3-Sulfopropyl)Quinolinium); Stilbene; Sulforodamine B Can C; Sulforodamine G Extra; SYTO11; SYTO12; SYTO13; SYTO14; SYTO15; SYT; SYTO17; SYTO18; SYTO20; SYTO21; SYTO22; SYTO23; SYTO24; SYTO25; SYTO40; SYTO41; SYTO42; SYTO43; SYTO44; SYTO45; SYTO59; SYTO60; SYTO61 ;SYTO62;SYTO63;SYTO64;SYTO80;SYTO81;SYTO82;SYTO83;SYTO84;SYTO85;SYTOX Blue;SYTOX Green;SYTOX Orange;Tetracycline;Tetramethylrhodamine (TRITC);Texas Red;Texas Red-X Conjugate;Thiadycarbocyanine (DiSC3);Thiazine Red R;Thiazole Orange;Thioflavin 5;Thioflavin S;Thioflavin TCN;Thiolite;Thiazole Orange;Chinopol CBS (Calcoflor White);TMR;TO-PRO-1;TO-PRO-3;TO-PRO-5;TOTO-1;TOTO-3;TriColor (PE-Cy5);TRITC (Tetramethylrhodamine-isothiocyanate);True Examples include Blue; TruRed; Ultralight; Uranine B; Uvitex SFC; wt GFP; WW781; X-Rhodamine; XRITC; Xylene Orange; Y66F; Y66H; Y66W; Yellow GFP; YFP; YO-PRO-1; YO-PRO-3; YOYO-1; and YOYO-3.

[0188] In one example, the detectable label is an enzyme. Examples of enzymes useful in this disclosure include, but are not limited to, alkaline phosphatase and horseradish peroxidase. Alternatively, or in addition, the enzyme may be, for example, luciferase. The enzyme can be ligated to an antibody by conventional chemical methods, or it can be expressed together with the antibody as a fusion protein. In one example, the enzyme is horseradish peroxidase.

[0189] The radioactive isotopes useful as detectable labels in this disclosure are well known in the art, 3 H, 11 C, 18 F, 35 S, 64 Cu, 67 Ga, 68 Ga, 99 mTC, 111 In, 123 I, 124 I, 125 I, and 131 I could include it.

[0190] Analysis of effector cells Methods for determining the proportion of basophil and / or mast cell activation and / or basophil and / or mast cell degranulation in this disclosure will be apparent to those skilled in the art and / or are described herein. For example, the proportions can be determined using ELISA, FLISA, or lateral flow assays. Briefly, basophils and / or mast cells are incubated with a plasma sample or fraction thereof for a suitable period of time. The 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 basophils and / or mast cells exposed to the sample is analyzed using flow cytometry.

[0191] As used herein, the term “percentage” relating to the activation of basophils and / or mast cells should be understood to refer to a measure of activation markers.

[0192] As used herein, the term “activation” should be understood to mean the stimulation of cells (e.g., basophils and / or mast cells) by exposure to an activation marker, such as a cell or soluble ligand, which results in a change in the cellular morphology or behavior.

[0193] For example, when a cell is activated, it upregulates activation markers. In some cases, the activation markers are selected from a group consisting of CD63, CD203c, CD107a, and combinations thereof.

[0194] As used herein, the term “degranulation” should be understood to mean the process by which cytoplasmic granules are released from a cell (e.g., mast cells and / or basophils).

[0195] In some cases, cell activation leads to degranulation. In other cases, cell activation does not lead to degranulation. For example, cell activation leads to increased cytokine production without degranulation.

[0196] In one example, basophil activation after exposure to a 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 measures the rate of basophil activation and / or degranulation using an activation marker. For example, the activation marker is CD63. In another example, the activation marker is CD203c. CD63 and CD203c are useful markers for flow cytometry quantification of in vitro activated basophils. In some examples, the basophils are activated human basophils.

[0197] In one example, the basophil activation rate is determined using flow cytometry. In another example, the basophil activation rate is determined by incubating basophils with a plasma sample or fraction thereof and quantifying the proportion of CD63-expressing basophils in the total basophil population using flow cytometry. In yet another example, the basophil activation rate is determined by incubating basophils with a plasma sample or fraction thereof and quantifying the proportion of CD203c-expressing basophils in the total basophil population using flow cytometry.

[0198] In one example, basophil activation is measured by the presence of interleukin-3 (IL-3). In another example, basophil activation is measured by the absence of IL-3.

[0199] In another example, mast cell activation after exposure to a plasma sample or fraction thereof can be determined using mast cell activation assays known in the art. For example, the assay is the Hoxb8 mast cell activation test. The assay measures the rate of mast cell activation and / or degranulation using an activation marker. For example, the activation marker is CD107a. CD107a is a useful marker for in vitro activated mast cells, quantified by flow cytometry. In some examples, the mast cells are Hoxb8 mast cells.

[0200] In one example, the rate 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 entire mast cell population using flow cytometry.

[0201] In short, basophils or mast cells are incubated with a plasma sample or fraction thereof. Cell activation is detected by detecting the percentage of cells expressing activation markers (e.g., CD63, CD203c, and / or CD107a). The activation rate (i.e., the percentage of cells expressing the activation marker in the entire cell population) is then quantified using flow cytometry.

[0202] From the disclosure herein, it will be clear to those skilled in the art that the method for determining the activation and / or degranulation of basophils and / or mast cells is performed in vitro.

[0203] In one example, the method involves determining a detectable amount of anti-FcεRI Ig in a plasma sample or fraction by determining the percentage of basophils that express CD63 after exposure to the plasma sample or fraction. In one example, if at least 5% of the basophil population expresses CD63 after exposure to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction. For example, the percentage of basophils that express CD63 after exposure to the plasma sample or fraction is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of the total population of basophils exposed to the plasma sample or fraction.

[0204] In one example, the method involves determining a detectable amount of anti-FcεRI Ig in a plasma sample or fraction by determining the percentage of basophils that express CD203c after exposure to the plasma sample or fraction. In one example, if at least 5% of the basophil population expresses CD203c after exposure to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction. For example, the percentage of basophils that express CD203c after exposure to the plasma sample or fraction is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of the total population of basophils exposed to the plasma sample or fraction.

[0205] In one example, the method involves determining a detectable amount of anti-FcεRI Ig in a plasma sample or fraction by determining the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction. In one example, if at least 5% of the population of mast cells express CD107a after exposure to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction. For example, the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of the total population of mast cells exposed to the plasma sample or fraction.

[0206] Method for preparing an anti-FcεRI Ig reduction preparation This disclosure provides a method for preparing an anti-FcεRI Ig depletion preparation from plasma or a fraction thereof using a chromatographic resin. For example, this disclosure provides a method for preparing an anti-FcεRI Ig depletion preparation from plasma or a fraction thereof using an affinity chromatographic resin. The affinity resin of this disclosure comprises a ligand that can specifically bind to anti-FcεRI Ig.

[0207] As used herein, the terms “reduced” and “depleted” should be understood to mean an Ig preparation in which the amount of anti-FcεRI Ig is reduced compared to an Ig preparation in which anti-FcεRI Ig is not reduced or depleted, and is sufficient to prevent activation of the FcεRI-mediated signaling pathway. As will be understood by those skilled in the art through the disclosure herein, anti-FcεRI Ig does not need to be reduced or depleted by 100%, but only by an amount sufficient to prevent activation of the FcεRI-mediated signaling pathway.

[0208] As used herein, the terms “reduced basophil activation” and “reduced mast cell activation” should be understood to mean an Ig preparation in which the amount of anti-FcεRI Ig is reduced to decrease basophil or mast cell activation. As will be understood by those skilled in the art, reduced basophil activation or reduced mast cell activation does not necessarily refer to a 100% reduction in activation, but simply to a reduction compared to an Ig preparation in which anti-FcεRI Ig is not reduced or depleted.

[0209] Suitable affinity chromatography resins will be obvious to those skilled in the art and / or will be described herein. In one example, the resin comprises a ligand containing an FcεRI or a fragment or epitope thereof. In another example, the resin comprises a ligand containing an FcεRI antibody or a fragment thereof. Those skilled in the art will recognize that FcεRI-based ligands can specifically bind to all types of anti-FcεRI Ig (i.e., IgG and IgE).

[0210] Serial affinity chromatography In some cases, affinity chromatography is continuous affinity chromatography.

[0211] The term "continuous affinity chromatography" should be understood as a chromatography method involving one or more columns packed with the same affinity resin, each column containing one or more zones. A zone is a column or region of a column containing resin on which one or more chromatographic steps can be performed. For example, a zone may be selected from the group consisting of equilibrium zones, binding zones, washing zones, elution zones, exfoliation zones, or combinations thereof.

[0212] Serial affinity chromatography involving two or more columns involves columns connected in a configuration that allows the columns to be operated in series and / or in parallel. In principle, IgG may be loaded into the first and / or subsequent columns, while the other columns (or other zones of the columns) undergo equilibrium, washing, elution, and / or regeneration simultaneously. Examples of serial affinity chromatography will be obvious to those skilled in the art and / or will be described herein.

[0213] Examples of columns that may be used to carry out serial chromatography methods will be obvious to those skilled in the art and / or will be described herein. For example, a serial chromatography method may be carried out using Tricorn 5 / 100 (Cytiva). In another example, a serial chromatography method may be carried out using BioSMB PD System (Sartorius).

[0214] Simulated Moving Bed (SMB) Chromatography For example, continuous affinity chromatography is pseudo-mobile bed (SMB) chromatography. The terms “pseudo-mobile bed chromatography” or “SMB chromatography” refer to the chromatographic method first described in U.S. Patent No. 2,985,589. Examples of SMB chromatography setups and / or apparatus will be obvious to those skilled in the art and / or will be described herein. The concept of pseudo-mobile beds involves the use of multiple smaller columns (rather than one large column) containing a solid adsorbent (e.g., affinity resin) and performing one or more continuous chromatographic steps (i.e., equilibrium, binding, washing, elution, or exfoliation) simultaneously on different columns in a continuous loop.

[0215] An example of an SMB chromatography setup has columns arranged in four sections, each section having one or more columns. Two inlet flows (feed and eluent) and two outlet flows (extract and recovery) are directed to and from the column rings in an alternating sequence. The inlet and outlet positions are switched at regular time intervals in the direction of the liquid flow, thus simulating backflow movement of the column. The feed (containing adsorbent components (extract)) is loaded into one or more columns of the SMB chromatography setup, and the extract binds to the resin in the column. Meanwhile, lower adsorbent components in the feed (recovery) pass through the column. The recovery can be loaded into one or more subsequent columns or removed from the SMB chromatography system as waste. The eluent is loaded into the column to collect the extract. For example, the eluent may be collected from the first column while more feed is loaded into one or more subsequent columns.

[0216] Suitable washing and elution buffers having the features of this disclosure will be obvious to those skilled in the art and / or will be described herein. In one example, the washing buffer comprises 20 mM sodium dihydrogen phosphate, 145 mM sodium chloride, and has a pH of 7.4.

[0217] In SMB chromatography, the resin may undergo multiple cycles (e.g., 50 cycles) of resin equilibrium, IgG loading, binding, elution, detachment, sterilization, and / or regeneration for each batch of plasma sample or fraction used. Multiple batch runs (e.g., 4–10 batches) can be performed using SMB chromatography. The total lifespan of the resin in SMB chromatography can range from 200–500 cycles (if not more) before the resin becomes unusable. Resin regeneration is typically performed to allow for multiple uses of the resin.

[0218] Periodic countercurrent chromatography (PCC) One example of continuous affinity chromatography is periodic countercurrent chromatography (PCC). Examples of PCC setups and / or apparatus will be obvious to those skilled in the art and / or will be described herein. The concept of PCC involves using multiple columns containing solid adsorbents (e.g., affinity resins) and performing chromatographic steps in parallel by semi-continuous means. The buffers used in the binding, washing, and / or elution steps flow countercurrently with respect to the affinity resin.

[0219] An example of a PCC setup involves the use of two columns. In the first step, a sample is loaded onto the first column on a resin DBC so that unbound products (e.g., IgG) pass through the first column and are captured by the second column. In the second step, the first column is washed, eluted, purified, and / or re-equilibrated independently of the second column, which is loaded with further sample. In the third step, additional sample is loaded onto the second column on a resin DBC so that unbound products pass through the second column and are captured by the first column. In the fourth step, the second column is washed, eluted, purified, and / or re-equilibrated independently of the first column, which is loaded with further sample. The process steps are cycled sequentially between the two columns.

[0220] Another example of a PCC configuration involves the use of multiple columns. For example, variations of the above PCC configuration could involve the use of multiple columns to capture unbound products, simulating the use of a large column.

[0221] Continuous countercurrent tangential chromatography (CCTC) One example of continuous affinity chromatography is continuous countercurrent tangential chromatography (CCTC). Examples of CCTC setups and / or apparatus will be obvious to those skilled in the art and / or will be described herein. The concept of CCTC involves using affinity resins in slurry form, where the slurry is continuously guided through a number of static mixers and a hollow fiber membrane that separates the fluid phase from the resin. CCTC is typically performed at low pressure (e.g., less than 70 kPa).

[0222] An example of a CCTC process includes binding, first washing, second washing, elution, detachment, and / or equilibration steps. The sample (e.g., plasma sample or fraction thereof) and affinity resin pass through a static mixer and a holofiber membrane in the binding step. Impurities are removed by flow-through of the holofiber membrane in the washing step, while the resin-bound product (i.e., IgG) is held by the membrane. The holofiber holds the resin and allows the product to flow through in the elution step. The resin is detached and / or equilibrated, and the process is repeated.

[0223] Continuous countercurrent spiral chromatography (CCSC) One example of continuous affinity chromatography is continuous countercurrent spiral chromatography (CCSC). Examples of CCSC setups and / or apparatus will be obvious to those skilled in the art and / or will be described herein. The concept of CCSC involves the use of a compact rotary coil separation column mounted on the rotary frame of a centrifuge. Two separation column designs are currently available: spiral disk assemblies and spiral tube support assemblies.

[0224] An exemplary CCSC process involves a coiled separation column that rotates around the central axis of a centrifuge, which rotates synchronously around its own axis (e.g., at 1,000–1,200 rpm). The mobile phase can pass through the centrifuge rotor without a rotary seal, and a large amount of stationary phase is held, while the two phases are mixed along the length of the column, resulting in highly efficient solute separation.

[0225] buffer solution This disclosure provides an affinity chromatography method using a buffer that enables efficient binding of Ig to a resin and collection from the resin. Generally, the plasma sample or fraction thereof is at a neutral pH (pH about 7.4). The resin is equilibrated with an equilibrium buffer and / or washed with a wash buffer having a buffer range covering a neutral pH. A suitable wash buffer contains a buffer having a dissociation constant (pKa) between 6.8 and 8.5 at 25°C.

[0226] An exemplary buffer for equilibrium buffers and / or wash buffers is sodium dihydrogen phosphate, the phosphate component of which has three dissociation constants (pKa: 2.16, 7.21, and 12.32). Phosphate has a dissociation constant near the pH of the elution buffer and / or exfoliation buffer used in continuous affinity chromatography. However, phosphate does not have a dissociation constant between the pH of the equilibrium buffer and / or wash buffer (higher pH) and the pH of the elution buffer and / or exfoliation buffer (lower pH) used in affinity chromatography. This allows for rapid switching between the wash and elution steps, as well as the exfoliation and equilibration steps, resulting in more defined peaks and shorter chromatographic phases. An advantage of using such equilibrium buffers and / or wash buffers is that smaller buffer volumes can be used, thereby increasing the efficiency of affinity chromatography.

[0227] Other suitable buffers for equilibrium buffers and / or wash buffers 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), and TEA (pKa: 7.76).

[0228] Chromatographic resin This disclosure provides a method for preparing anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparations from a plasma sample or fraction thereof using a continuous chromatography resin. In one example, the resin of this disclosure contains a ligand that can specifically bind to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig).

[0229] In one example, the method further includes the use of affinity chromatography resin to remove anti-A and anti-B antibodies. In another example, the method further includes the use of affinity chromatography resin to remove anti-A antibody. In yet another example, the method further includes the use of affinity chromatography resin to remove anti-B antibody.

[0230] Suitable chromatography resins will be obvious to those skilled in the art and / or will be described in US2009 / 074749, which is incorporated herein by reference. In one example, the resin comprises a matrix of a support grafted with oligosaccharide groups having antigenic similarity to blood types A and B. An exemplary resin is Glycosorb Abo® (Glycorex Transplantation AS).

[0231] Additional purification steps As will be understood by those skilled in the art, additional steps can be performed before or after the continuous chromatography step. In one example, the additional step can be performed before the continuous chromatography step. In one example, the additional step can be performed after the continuous chromatography step.

[0232] In one example, the method further comprises one or more steps selected from the group consisting of ethanol precipitation, octanoic acid fractionation, ion exchange chromatography, virus inactivation, virus filtration, and ultrafiltration / diafiltration. Additional purification steps will be apparent to those skilled in the art and / or will be described herein.

[0233] In one example, the method further comprises ethanol precipitation. For example, using chilled ethanol to remove albumin and α- and β-globulins from a plasma sample or a fraction thereof, an anti-FcεRI Ig-reduced preparation can be isolated and concentrated. For example, as described in WO2011 / 149472.

[0234] In one example, the method further comprises octanoic acid fractionation. Octanoic acid can be used to remove plasma lipids and plasma proteins (other than IgG). For example, as described in WO2011 / 131787.

[0235] In one example, the method further comprises ion exchange chromatography. For example, anion exchange chromatography can be used to remove IgA, residual IgM, and other plasma components (other than IgG).

[0236] The anion exchanger can be a resin-based anion exchanger, an anion exchange membrane adsorber, or any other format of anion exchanger having 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 an integrated anion exchanger.

[0237] In one example, the method further includes 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 includes a matrix composed of a poly(styrene-divinylbenzene) matrix. In one example, the strong anion exchanger includes a quaternized polyethyleneimine functional group. Suitable resin-based anion exchangers will be apparent to those skilled in the art and include, for example, POROS™ HQ50.

[0238] 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 binding and elution mode.

[0239] In one example, the anion exchange chromatography step includes a buffer selected from the group consisting of sodium citrate, 2-(N-morpholino)ethanesulfonic acid (MES) buffer, sodium dihydrogen phosphate, bis-tris, phosphoric acid, L-histidine, and combinations thereof. In one example, the anion exchange chromatography step includes a buffer that includes an MES buffer. In another example, the anion exchange chromatography step includes a phosphate buffer.

[0240] In one example, the method further includes virus inactivation. For example, virus inactivation can be achieved by adjusting the solution to a low pH. The low pH can be a pH of 2-4. In one example, the low pH virus inactivation is performed in the presence of caprylate. In another example, virus inactivation can be achieved by contacting a plasma sample or a fraction thereof, or a pharmaceutical composition containing an anti-FcεRI Ig-reduced preparation or an anti-FcεRI Ig-reduced preparation with n-octyl-β-D-glucopyranoside (OG), thereby forming an OG-IgG mixture. In a further example, the low pH virus inactivation is performed in the presence of N,N-dimethylmyristylamine N-oxide (TDAO).

[0241] In further examples, viral inactivation may be achieved by exposing a protein mixture, plasma or plasma fraction, protein-reduced preparation or composition (e.g., plasma protein-reduced preparation, e.g., IgG-reduced preparation) to a solvent-detergent inactivation step. Suitable solvent-detergent treatments will be apparent to those skilled in the art and include, for example, environmentally friendly detergents. Exemplary environmentally friendly detergents suitable for use in this disclosure and particularly for use in the inactivation of lipid-enveloped viruses include N,N-dimethylmyristylamine N-oxide (TDAO), polysorbate 80 (PS80), polyoxyethylene (10) isooctylcyclohexyl ether (TRITON® X-100-reduction), and nonionic surfactants prepared from glucose and alcohol (e.g., Simulsol® formulations). In one example, the detergent is N,N-dimethylmyristylamine N-oxide (TDAO). In another example, the detergent is polysorbate 80. In another example, the detergent is polyoxyethylene(10) isooctylcyclohexyl ether (TRITON® X-100-reduction). In yet another example, the detergent is a nonionic surfactant prepared from glucose and alcohol.

[0242] In one example, the method further includes viral filtration. For instance, a viral filtration membrane with a pore size of 1–20 nm may be used to remove microorganisms and viruses from a solution, eluent, or pharmaceutical composition. An exemplary nanofilter is Planova 20 (Planova). Examples of nanofilters include Planova S20N (Asahi), Virosart HC (Sartorius), and Planova 20N (Asahi).

[0243] In one example, the method further includes ultrafiltration / diafiltration. Illustrative ultrafiltration / diafiltration membranes include Pellicon 2 cassettes (Millipore) or polyethersulfone or Hydrosart cassettes (Sartorius).

[0244] Analysis of anti-FcεRI Ig reduction preparations In one example, the activity of an anti-FcεRI Ig reduction preparation is evaluated. Methods for determining the yield, purity, and IgG subclass distribution will be apparent to those skilled in the art and / or will be described herein.

[0245] In one example, purity is determined by SDS-PAGE and MALDI-TOF-MS peptide fingerprint analysis. Briefly, the purified plasma protein product or pharmaceutical composition described herein is loaded onto a suitable SDS-PAGE gel (e.g., 8-16% TRIS-glycine) under reducing and non-reducing conditions, along with a protein size marker and a positive control of the protein of interest (e.g., IgG, e.g., IVIG). The proteins are separated based on size, and the protein band of interest is isolated, processed, and analyzed by MALDI-TOF-MS.

[0246] In another example, impurities in the anti-FcεRI Ig-reducing preparations or pharmaceutical compositions described herein are measured by enzyme-linked immunosorbent assay (ELISA) using an impurity (e.g., IgA) specific antibody. For example, ELISA is performed using commercially available methods.

[0247] In one example, the purity, yield, and / or subclass distribution of IgG are determined by diffuse light measurements.

[0248] In one example, the anti-FcεRI Ig reduction preparation contains Ig with a purity of over 95%. For example, the anti-FcεRI Ig reduction preparation contains Ig with a purity of over 96%. In another example, the anti-FcεRI Ig reduction preparation contains Ig with a purity of over 97%. In yet another example, the anti-FcεRI Ig reduction preparation contains Ig with a purity of over 98%. In yet another example, the anti-FcεRI Ig reduction preparation contains Ig with a purity of over 99%.

[0249] Stability of plasma and plasma fractions The stability of plasma samples or fractions for loading into the chromatographic resins described herein can be determined by evaluating the procoagulant activity, proteolytic activity, and particle size of the plasma samples or fractions. Methods for evaluating procoagulant activity, proteolytic activity, and particle size will be apparent to those skilled in the art and / or will be described herein. Briefly, plasma or plasma fractions are frozen / thawed for one or more cycles, stored at 2°C to 32°C (e.g., 2°C, 10°C, 18°C, 21°C, 28°C, or 32°C) for 24 hours or up to 48 hours, and analyzed using one or more of the methods described below. In one example, a plasma sample or fraction is thawed for one or more cycles at 32°C, stored for 24 hours or up to 48 hours, and analyzed using one or more of the methods described below. In another example, a plasma sample or fraction is thawed for one or more cycles at 32°C, stored for 24 hours or up to 48 hours, analyzed using one or more of the methods described below, then cooled and stored at 21°C. In one example, the plasma sample or its fraction is thawed at 32°C and 21°C before serial chromatography.

[0250] For example, the procoagulation activity in a plasma sample or fraction can be determined using an in vitro coagulation assay, such as the activated partial thromboplastin time (NaPTT) assay. The NaPTT assay measures the rate at which one or more coagulation factors (e.g., fibrinogen, prothrombin, proaccelerin, antihemophilic factor, Stuart-Prower factor, plasma thromboplastin precursor, and Hegeman factor) are activated or formed in a plasma sample or fraction when a coagulation activator (e.g., silica, kaolin, ellagic acid) is added to the assay.

[0251] For example, the proteolytic activity in a plasma sample or fraction thereof can be evaluated by measuring the activity of thrombin, common serine proteases, kallikrein, plasmin, and FXa using commercially available kits such as the thrombin activity assay kit (S-2238), common serine protease assay kit (S-2288), kallikrein activity assay kit (S-2302), plasmin activity assay kit (S-2251), and FXa activity kit (S-2765).

[0252] In one example, the size of any particles in a plasma sample or fraction thereof is assessed by microflow imaging (MFI), and a polydispersity index is calculated. The calculation of the polydispersity index will be obvious to those skilled in the art.

[0253] Pharmaceutical composition The anti-FcεRI Ig-decreasing plasma preparations and anti-FcεRI Ig-decreasing IgG preparations of this disclosure are useful for formulation into pharmaceutical compositions for parenteral use, such as intravenous or subcutaneous administration, for therapeutic and prophylactic treatment.

[0254] A preferred embodiment of the present invention comprises an anti-FcεRI Ig-decreasing preparation or an anti-FcεRI Ig-decreasing IgG plasma preparation.

[0255] The composition for administration generally comprises a solution of the anti-FcεRI Ig-decreasing preparation or anti-FcεRI Ig-decreasing IgG preparation of this disclosure, dissolved in a pharmaceutically acceptable carrier such as an aqueous carrier. Various aqueous carriers, such as buffered saline, can be used. The composition may contain pharmaceutically acceptable carriers required to approximate physiological conditions, such as agents for adjusting pH and buffering, as well as agents for adjusting toxicity, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, and sodium lactate.

[0256] The concentration of Ig in the pharmaceutical compositions of the present disclosure in these formulations can vary widely and will be selected primarily based on fluid volume, viscosity, body weight, etc., according to the particular mode of administration selected and the needs of the patient. The vehicle may contain small amounts of additives to enhance isotonicity and chemical stability, such as buffers and preservatives. For example, the pharmaceutical composition contains proline as a stabilizer.

[0257] Suitable pharmaceutical compositions according to the present disclosure generally include an amount of the anti-FcεRI Ig-reducing preparation of the present disclosure mixed with an acceptable pharmaceutical carrier, such as a sterile aqueous solution, to obtain 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.

[0258] For example, the total Ig concentration of the pharmaceutical composition is 1 - 5% w / v, 5 - 15% w / v, or 8 - 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., 10 g Ig / L) can be used. For intravenous use, 10% w / v (i.e., 100 g Ig / L) can be used.

[0259] For subcutaneous administration, higher concentrations can be used. For example, 15 - 35% w / v, or 20 - 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.

[0260] Anti-FcεRI Ig-reducing Ig preparation This disclosure provides anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) reduction preparations containing polyclonal IgG.

[0261] This disclosure also provides anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion IgG preparations, which include polyclonal IgG.

[0262] In one example, polyclonal IgG is present in an amount of 5-25% (w / v). In another example, polyclonal IgG is present in an amount of 6-15% (w / v). In yet another example, polyclonal IgG is present in an amount of approximately 8-12% (w / v). In yet another example, polyclonal IgG is present in an amount of approximately 10% (w / v).

[0263] The final protein concentration will depend on various factors such as the route of administration and the type of condition being treated. Those skilled in the art will be able to determine the optimal protein concentration for the intended application.

[0264] For example, in intravenous infusion, polyclonal IgG is present in an amount of approximately 15-20% (w / v). In one example, polyclonal IgG is present in an amount of approximately 8-12% (w / v). In another example, polyclonal IgG is present in an amount of approximately 20% (w / v).

[0265] For subcutaneous administration, higher doses, such as approximately 15-20% (w / v), may be selected. In one example, polyclonal IgG is present in an amount of approximately 20% (w / v).

[0266] Immunoglobulins can be isolated from human or animal blood, or produced by other means, such as recombinant DNA technology or hybridoma technology. For example, immunoglobulins are obtained from blood plasma, typically from a pool of blood plasma from multiple donors. To obtain immunoglobulins from plasma, the plasma is subjected to alcohol fractionation and may be combined with other purification techniques such as chromatography, adsorption, or precipitation as described herein. However, other processes may also be used.

[0267] The pharmaceutical compositions of this disclosure are formulated by methods known in the art. In the case of IgG solutions, the pH of the final preparation is adjusted to a relatively high but acidic pH, i.e., in the range of approximately pH 4.2 to 5.4. This pH range has been found to be particularly useful for improving the preservation of the properties of the polyclonal IgG preparation. In one example, the pH is approximately 4.6 to 5.0. In another example, the pH is 4.8.

[0268] In other examples, the pharmaceutical composition further includes a stabilizer. In one example, the stabilizer is two or more amino acids. For example, the amino acids are selected from the group consisting of nonpolar and basic amino acids. In some examples, the amino acids are selected from the group consisting of histidine, arginine, lysine, ornithine, isoleucine, valine, methionine, glycine, and proline.

[0269] In one example, the stabilizer is proline. For example, proline is L-proline. The amount of proline in the pharmaceutical composition ranges from approximately 10 to approximately 2000 mmol / l. In some examples, the amount of proline in the pharmaceutical composition ranges from approximately 50 to approximately 1000 mmol / l. In other examples, the amount of proline in the pharmaceutical composition ranges from approximately 100 to approximately 500 mmol / l.

[0270] For example, the amount of L-proline in a pharmaceutical composition is in the range of approximately 200 mmol / L to 300 mmol / L. For instance, the amount of L-proline in a preparation is in the range of approximately 225 mmol / L to 275 mmol / L. For example, the amount of L-proline in a pharmaceutical composition is in the range of approximately 240 mmol / L to 260 mmol / L. For instance, the amount of L-proline in a pharmaceutical composition is in the range of approximately 250 mmol / L.

[0271] In a further example, the amount of proline in a pharmaceutical composition is approximately 250 mmol / l.

[0272] In one example, the final proline concentration is 200 mM to 400 mM. In another example, the final proline concentration is 250 mM.

[0273] In one example, proline is L-proline. In another example, proline is a proline equivalent (e.g., a proline analog).

[0274] In other examples, the stabilizer is present in the pharmaceutical composition at a concentration greater than 200 mM. In some examples, the stabilizer is present in the pharmaceutical composition at a concentration of 200 mM to 400 mM. In yet another example, the stabilizer is present in the pharmaceutical composition at a concentration of 200 mM to 300 mM. In yet another example, the stabilizer is present in the pharmaceutical composition at a concentration of 250 mM.

[0275] In one example, the pharmaceutical composition contains a pH of 4 to 5.5. For example, the pharmaceutical composition contains a pH of 4.5 to 5.0. In another example, the pharmaceutical composition contains a pH of 4.6 to 5.0. For example, the pharmaceutical composition contains a pH of 4.6. In one example, the pharmaceutical composition or Ig preparation or anti-FcεRI Ig reduction preparation contains a pH of 4.7. In yet another example, the pharmaceutical composition contains a pH of 4.8. In yet another example, the pharmaceutical composition contains a pH of 4.9. In one example, the pharmaceutical composition contains a pH of 5.0.

[0276] In one example, the pharmaceutical composition contains 100 mg / mL of total human plasma protein. In another example, the pharmaceutical composition contains 20 g / 100 mL of total human plasma protein.

[0277] In one example, the pharmaceutical composition contains immunoglobulin G (IgG) with a purity of over 95%. For example, the pharmaceutical composition contains immunoglobulin G (IgG) with a purity of over 96%. In another example, the pharmaceutical composition contains immunoglobulin G (IgG) with a purity of over 97%. In yet another example, the pharmaceutical composition contains immunoglobulin G (IgG) with a purity of over 98%. In yet another example, the pharmaceutical composition contains immunoglobulin G (IgG) with a purity of over 99%.

[0278] For example, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain a distribution of more than 60% IgG1 subclasses.

[0279] For example, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain less than 30% of the IgG2 subclass distribution. For instance, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain less than 28% of the IgG2 subclass distribution.

[0280] For example, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain less than 5% of the IgG3 subclass distribution.

[0281] For example, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain less than 5% of the IgG4 subclass distribution. For instance, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain less than 3% of the IgG4 subclass distribution.

[0282] For example, a pharmaceutical composition or anti-FcεRI Ig-reducing preparation may contain an IgG subclass distribution similar to that of normal human plasma, e.g., 69% IgG1, 26% IgG2, 3% IgG3, and 2% IgG4.

[0283] For example, the pharmaceutical composition contains a nominal osmotic pressure of approximately 300 mOsm / kg to 400 mOsm / kg. For example, the pharmaceutical composition contains a nominal osmotic pressure of 380 mOsm / kg. For example, the pharmaceutical composition contains a nominal osmotic pressure of approximately 300 mOsm / kg to 350 mOsm / kg. For example, the pharmaceutical composition contains a nominal osmotic pressure of 320 mOsm / kg.

[0284] For example, the pharmaceutical composition contains a sodium content of 1 mmol / L or less.

[0285] For example, the pharmaceutical composition contains an IgA content of 0.05 mg / mL or less. For example, the pharmaceutical composition contains an IgA content of 0.04 mg / mL or less, or 0.03 mg / mL or less. For example, the pharmaceutical composition contains an IgA content of 0.025 mg / mL or less. For example, the pharmaceutical composition contains an IgA content of 0.01 mg / mL or less. For example, the pharmaceutical composition contains an IgA content of 0.009 mg / mL or less.

[0286] For example, the pharmaceutical composition contains an IgA content of 0.1 mg / g IgG or less.

[0287] For example, the pharmaceutical composition contains an IgA content of 0.09 mg / g IgG or less.

[0288] For example, a pharmaceutical composition contains an IgM content of 10 mg / L or less. For example, IgM content of 10 mg / L or less, 9 mg / L or less, 8 mg / L or less, 7 mg / L or less, 6 mg / L or less, 5 mg / L or less, 4 mg / L or less, 3 mg / L or less, or 2 mg / L or less. For example, a pharmaceutical composition contains an IgM content of 2 mg / L or less. For example, a pharmaceutical composition contains an IgM content of 1 mg / L or less. For example, a pharmaceutical composition contains an IgM content of 0.5 mg / L or less. For example, a pharmaceutical composition contains an IgM content of less than 0.17 mg / L.

[0289] For example, the pharmaceutical composition contains an IgM content of 2 μg / g IgG or less.

[0290] For example, the pharmaceutical composition contains an IgM content of 1.9 μg / g IgG or less.

[0291] For example, the pharmaceutical composition contains an albumin content of 0.50 mg / mL or less. For example, the pharmaceutical composition contains an albumin content of 0.40 mg / mL or less. For example, the pharmaceutical composition or anti-FcεRI Ig-reducing Ig preparation contains an albumin content of 0.30 mg / mL or less. For example, the pharmaceutical composition contains an albumin content of 0.20 mg / mL or less. For example, the pharmaceutical composition contains an albumin content of 0.10 mg / mL or less. For example, the pharmaceutical composition contains an albumin content of 0.09 mg / mL or less. For example, the pharmaceutical composition contains an albumin content of 0.08 mg / mL or less. For example, the pharmaceutical composition contains an albumin content of 0.07 mg / mL or less.

[0292] In one example, the pharmaceutical composition contains an albumin content of 1 mg / g IgG or less. In another example, the pharmaceutical composition contains an albumin content of 0.80 mg / g IgG or less.

[0293] For example, the pharmaceutical composition contains a prekallikrein activator (PKA) level of 35 IU / mL or less. For example, the pharmaceutical composition contains a prekallikrein activator (PKA) level of 30 IU / mL or less. For example, the pharmaceutical composition contains a prekallikrein activator (PKA) level of 50 IU / mL or less. For example, the pharmaceutical composition contains a prekallikrein activator (PKA) level of 20 IU / mL or less. For example, the pharmaceutical composition contains a prekallikrein activator (PKA) level of 15 IU / mL or less. For example, the pharmaceutical composition contains a prekallikrein activator (PKA) level of 10 IU / mL or less.

[0294] For example, an anti-FcεRI Ig reduction preparation contains 10% (w / v) polyclonal IgG, proline at a concentration of 250 mM, and a pH of 4.8.

[0295] For example, an anti-FcεRI Ig reduction preparation contains 20% (w / v) polyclonal IgG, 250 mM proline, 20 μg / mL PS80, and a pH of 4.8.

[0296] An exemplary method for purifying the Ig process is described in WO2015 / 000886, and an exemplary IgG product is described in WO2016 / 087569, both of which are incorporated herein by reference.

[0297] This disclosure also provides a pooled IgG preparation containing a detectable amount of anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig). Those skilled in the art will see from this disclosure that a pooled IgG preparation containing a detectable amount of anti-FcεRI Ig is not provided by any of the methods of this disclosure. Therefore, this disclosure also provides a method for detecting anti-FcεRI Ig in a pooled IgG preparation.

[0298] In one example, the pooled IgG preparation contains IgA and other isotypes such as IgM. In another example, the pooled IgG preparation contains IgA. In yet another example, the pooled IgG preparation contains other isotypes such as IgM.

[0299] How to use As discussed herein, the disclosure provides methods for treating, preventing, and / or delaying the progression of conditions associated with primary or secondary immunodeficiency, inflammatory diseases, autoimmune diseases, or acute infections in subjects, the methods comprising administering to subjects an anti-FcεRI Ig-decreasing preparation, an anti-FcεRI Ig-decreasing IgG preparation, or a pharmaceutical formulation.

[0300] This disclosure also provides anti-FcεRI Ig-decreasing preparations or anti-FcεRI Ig-decreasing IgG preparations or the pharmaceutical compositions described herein for use in treating, preventing, and / or delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects.

[0301] This disclosure further provides anti-FcεRI Ig-decreasing preparations or anti-FcεRI Ig-decreasing IgG preparations or pharmaceutical compositions described herein for the manufacture of pharmaceuticals for treating, preventing, and / or delaying the progression of conditions associated with immunodeficiency, inflammatory diseases, autoimmune diseases, and / or acute infections in subjects.

[0302] In one example, this condition is selected from a group consisting of primary immunodeficiency (PI), chronic inflammatory demyelinating polyneuropathy (CIDP), chronic immune thrombocytopenic purpura (ITP), and combinations thereof.

[0303] In one example, the condition is primary immunodeficiency (PI).

[0304] In one example, the condition is chronic inflammatory demyelinating polyneuropathy (CIDP).

[0305] In one example, the condition is chronic immune thrombocytopenic purpura (ITP).

[0306] Other examples of conditions include congenital agammaglobulinemia and hypogammaglobulinemia, unclassifiable immunodeficiency, severe combined immunodeficiency, allogeneic bone marrow transplantation, chronic lymphocytic leukemia, childhood HIV, kidney transplantation with high antibody recipients or ABO-incompatible donors, chronic fatigue syndrome, and Clostridium. The following conditions are selected from the group consisting of 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, kidney transplant rejection, spontaneous abortion, generalized rigidity 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 deficiency due to recurrent infection, and combinations thereof.

[0307] In further examples, treatment may include replacement therapy in myeloma or chronic lymphocytic leukemia with severe secondary hypogammaglobulinemia and recurrent infections.

[0308] In other cases, the condition is selected from a group consisting of autoimmune diseases and certain neurological disorders, such as rheumatoid arthritis, systemic lupus erythematosus (SLE), antiphospholipid syndrome, immune thrombocytopenia (ITP), Kawasaki disease, Guillain-Barré syndrome (GBS), multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CIDP), multifocal motor neuropathy (MMN), myasthenia gravis (MG), cutaneous bullous diseases, scleroderma, dermatomyositis, polymyositis, Alzheimer's disease, Parkinson's disease, Alzheimer's disease associated with Down syndrome, cerebral amyloid angiopathy, Lewy body dementia, frontotemporal dementia, vascular dementia, cell and organ transplantation, and combinations thereof.

[0309] In some cases, the pharmaceutical composition is present in a vial, pre-filled syringe, or auto-injector device.

[0310] This disclosure also provides pre-filled syringes containing the pharmaceutical compositions described herein.

[0311] This disclosure also provides an auto-injector device comprising a pharmaceutical composition described herein.

[0312] In one example, the compositions of the present disclosure are administered subcutaneously to subjects requiring them. In another example, the compositions of the present disclosure are administered intravenously to subjects requiring them.

[0313] In one example, the compositions of this disclosure are self-administered.

[0314] In one example, the compositions of this disclosure are administered subcutaneously by the patient.

[0315] In one example, the composition of the present disclosure is provided in a pre-filled syringe.

[0316] In one example, the composition of this disclosure is administered subcutaneously by the patient using a pre-filled syringe.

[0317] In one example of any method described herein, the subject is a mammal, such as a primate, including humans.

[0318] kit Another example of the present disclosure is the provision of a kit comprising an FcεRI or a fragment or epitope thereof for use in any of the methods described herein (e.g., detecting anti-FcεRI Ig in a plasma sample or fraction thereof).

[0319] For example, the panels or kits described herein are for ex vivo analysis. For example, the kits are suitable for use with whole blood samples, plasma samples, or fractions thereof.

[0320] Reagents for specific types of assays can also be provided in kits. For example, a kit may include a device such as a lateral flow assay device, an analytical rotor, or an electrochemical, optical, or photoelectronic sensor, or a plate (e.g., a plate suitable for an ELISA assay). In examples where the kit includes a lateral flow assay device, the ligand (e.g., FcεRI or a fragment or epitope thereof) may be contained in a separate kit from the device, or it may be contained within the device itself, for example, it may be dried on a conjugate area within the device.

[0321] In addition, the kit includes various diluents and buffers, labeled conjugates or other agents for carrying out the above method, and other signal-generating reagents such as enzyme substrates, cofactors, and chromogens. Other components of the kit can be readily determined by those skilled in the art. Such components may include coating reagents, indicator charts for colorimetric comparison, disposable gloves, decontamination instructions, applicator sticks or containers, sample preparation cups, etc. In one example, the kit includes buffers or other reagents suitable for constituting a reaction medium in which the compositions disclosed herein come into contact with the sample.

[0322] In one example, the kit further includes instructions or a package insert having instructions for detecting and / or screening anti-FcεRI Ig in a plasma sample or fraction thereof. For example, in a particular example, the kit includes instructions showing how to use the kit to detect anti-FcεRI Ig, or how to screen and / or identify plasma samples suitable for administration to a subject, or how to identify subjects suitable and / or unsuitable for plasma donation for the manufacture of Ig preparations. In one example, the kit includes instructions showing how to prepare a sample. In one example, the kit provides instructions for contacting a sample with ligands disclosed herein (i.e., FcεRI or fragments or epitopes) in any order before analyzing the sample for the presence of anti-FcεRI Ig. The kit may also provide instructions for optimizing buffers, optimizing ratios of various components, optimizing sample dilutions, and optimizing the order of mixtures and application steps (e.g., mixing all components before application, mixing only specific components and applying others separately).

[0323] The present invention is further disclosed in the following numbered paragraphs: 1. A method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, wherein the method is (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0324] 2. A method for screening plasma samples or fractions for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), wherein the method is (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0325] 3. A method for identifying a plasma sample or fraction suitable for administration to a target, wherein the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0326] 4. A method for screening plasma samples or fractions thereof to determine their suitability for administration to a subject, wherein the method is: (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0327] 5. A method for identifying a plasma sample or fraction thereof for use in preparing an immunoglobulin (Ig) preparation for administration to a subject, wherein the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI)Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0328] 6. A method for screening plasma samples or fractions thereof for use in the preparation of immunoglobulin (Ig) preparations for administration to a subject, wherein the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI)Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0329] 7. The method according to any one of paragraphs 1 to 6, further comprising administering a plasma sample or fraction thereof to a subject if anti-FcεRI Ig is not present in a detectable amount.

[0330] 8. The method according to any one of paragraphs 1 to 8, further comprising including a plasma sample or fraction thereof in a pooled blood plasma sample if anti-FcεRI Ig is not present in a detectable amount.

[0331] 9. The method according to any one of paragraphs 1 to 8, further comprising excluding a plasma sample or fraction thereof from a pooled blood plasma sample if anti-FcεRI Ig is present in a detectable amount.

[0332] 10. The method according to any one of paragraphs 1 to 9, further comprising contacting the complex with a detection protein.

[0333] 11. The method according to paragraph 10, wherein the detected protein contains a detectable label, and the presence of the detectable label indicates the presence of a detectable amount of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0334] 12. The method according to paragraph 10 or 11, wherein the detected protein includes an antibody variable region that binds to anti-FcεRI Ig.

[0335] 13. The method according to paragraph 10, further comprising contacting a detection protein with an antibody containing a label that binds to it and is detectable.

[0336] 14. A method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, wherein the method is (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0337] 15. A method for screening plasma samples or fractions for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), wherein the method is (i) Contacting the sample with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0338] 16. A method for identifying a plasma sample or fraction suitable for administration to a target, wherein the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is not suitable for administration to a subject.

[0339] 17. A method for screening a plasma sample or fraction thereof to determine its suitability for administration to a subject, wherein the method is: (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is not suitable for administration to a subject.

[0340] 18. A method for identifying a plasma sample or fraction thereof for use in preparing an immunoglobulin (Ig) preparation for administration to a subject, wherein the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a target.

[0341] 19. A method for screening plasma samples or fractions thereof for use in the preparation of immunoglobulin (Ig) preparations for administration to a subject, wherein the method is (i) Contacting the sample with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a target.

[0342] 20. The method according to any one of paragraphs 14-19, further comprising contacting the complex with a detection protein.

[0343] 21. The method according to any one of paragraphs 14-20, wherein the detected protein includes an antibody variable region that binds to anti-FcεRI Ig.

[0344] 22. The method according to any one of paragraphs 1 to 21, wherein the ligand is immobilized on a solid surface.

[0345] 23. The method according to any one of paragraphs 1 to 22, further comprising immobilizing a ligand on a solid surface.

[0346] 24. A method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0347] 25. A method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0348] 26. A method for screening plasma samples or fractions for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0349] 27. A method for screening plasma samples or fractions thereof for anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig), wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0350] 28. A method for identifying a plasma sample or fraction thereof suitable for administration to a subject, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0351] 29. A method for identifying a plasma sample or fraction thereof suitable for administration to a subject, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0352] 30. A method for screening a plasma sample or fraction thereof to determine its suitability for administration to a subject, wherein the method is: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0353] 31. A method for screening a plasma sample or fraction thereof to determine its suitability for administration to a subject, wherein the method is: (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for administration to a subject.

[0354] 32. A method for identifying a plasma sample or fraction thereof for use in preparing an immunoglobulin (Ig) preparation for administration to a subject, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0355] 33. A method for identifying a plasma sample or fraction thereof for use in preparing an immunoglobulin (Ig) preparation for administration to a subject, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in the plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0356] 34. A method for screening plasma samples or fractions thereof for use in the preparation of immunoglobulin (Ig) preparations for administration to a subject, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein is bound to anti-FcεRI Ig and the detection protein contains a detectable label. (iv) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0357] 35. A method for screening plasma samples or fractions thereof for use in the preparation of immunoglobulin (Ig) preparations for administration to a subject, wherein the method is (i) Immobilizing a ligand containing FcεRI or a fragment or epitope on a solid surface, (ii) Contacting a sample with a ligand immobilized on a solid surface, wherein the immobilized ligand binds to anti-FcεRI Ig in the sample, thereby forming a complex. (iii) Contacting the complex with a detection protein containing an antibody variable region, wherein the detection protein binds to anti-FcεRI Ig, (iv) Contacting the detection protein with an antibody containing a label that binds to it and is detectable, (v) A method comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the plasma sample or fraction thereof is unsuitable for use in preparing an Ig preparation for administration to a subject.

[0358] 36. The method according to any one of paragraphs 13-35, wherein if the label is not detected, anti-FcεRI Ig is not present in a detectable amount.

[0359] 37. The method according to paragraph 36, further comprising administering a plasma sample or fraction thereof to a subject if anti-FcεRI Ig is not present in a detectable amount.

[0360] 38. The method according to paragraph 36, further comprising including a plasma sample or fraction thereof in a pooled blood plasma sample if anti-FcεRI Ig is not present in a detectable amount.

[0361] 39. The method according to paragraph 36, further comprising excluding a plasma sample or fraction thereof from a pooled blood plasma sample if anti-FcεRI Ig is present in a detectable amount.

[0362] 40. A method for identifying suitable subjects for plasma donation for the manufacture of immunoglobulin (Ig) preparations, wherein the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-FcεRI Ig in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0363] 41. A method for screening subjects suitable for plasma donation for the manufacture of immunoglobulin (Ig) preparations, wherein the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0364] 42. The method according to paragraph 40 or 41, wherein the subject is suitable for plasma donation when anti-FcεRI Ig is not present.

[0365] 43. The method according to either paragraph 40 or 41, wherein the subject is not suitable for plasma donation if anti-FcεRI Ig is present.

[0366] 44. The method according to any one of paragraphs 40-43, further comprising contacting the complex with a detection protein.

[0367] 45. The method according to paragraph 44, wherein the detected protein includes an antibody variable region, and the detected protein binds to anti-FcεRI Ig.

[0368] 46. ​​The method according to paragraph 44 or 45, wherein the detected protein comprises a detectable label, and the presence of the detectable label indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0369] 47. The method according to paragraph 46, further comprising detecting a label, wherein the presence of a detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0370] 48. The method according to paragraph 44 or 45, further comprising contacting a detection protein with an antibody containing a label that binds to it and is detectable.

[0371] 49. A method for identifying suitable subjects for plasma donation for the manufacture of immunoglobulin (Ig) preparations, wherein the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the subject is suitable for plasma donation for the production of an Ig preparation.

[0372] 50. A method for screening subjects suitable for plasma donation for the manufacture of immunoglobulin (Ig) preparations, wherein the method is (i) Contacting a plasma sample or fraction from the subject with a ligand that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the sample, thereby forming a complex, (ii) A method comprising detecting a complex using a detection protein containing a detectable label, wherein the presence of the detectable label indicates the presence of anti-FcεRI Ig in a plasma sample or fraction thereof, and the presence of anti-FcεRI Ig in a plasma sample or fraction thereof indicates that the subject is suitable for plasma donation for the production of an Ig preparation.

[0373] 51. The method according to any one of paragraphs 40-50, wherein the ligand is immobilized on a solid surface.

[0374] 52. The method according to any one of paragraphs 40-51, further comprising immobilizing a ligand on a solid surface.

[0375] 53. The method according to any one of paragraphs 49 to 52, wherein the subject is suitable for plasma donation when anti-FcεRI Ig is not present in a detectable amount.

[0376] 54. The method according to any one of paragraphs 49-52, wherein the subject is not suitable for plasma donation if anti-FcεRI Ig is present in a detectable amount.

[0377] 55. The method according to any one of paragraphs 1 to 54, wherein the method is an enzyme-linked immunosorbent assay (ELISA).

[0378] 56. The method according to any one of paragraphs 1 to 54, wherein the method is a fluorescence-coupled immunosorbent assay (FLISA).

[0379] 57. The method described in any one of paragraphs 1 to 54, wherein the method is a lateral flow assay.

[0380] 58. The method according to any one of paragraphs 1 to 57, wherein anti-FcεRI Ig binds to an epitope of FcεRI and activates an FcεRI-mediated signaling pathway.

[0381] 59. The method according to any one of paragraphs 1-58, wherein anti-FcεRI Ig binds to the α-chain of FcεRI and activates the FcεRI-mediated signaling pathway.

[0382] 60. The method according to any one of paragraphs 1 to 59, wherein a detectable amount of anti-FcεRI Ig in a plasma sample or fraction thereof activates an FcεRI-mediated signaling pathway.

[0383] 61. The method according to paragraph 59 or 60, wherein activation of the FcεRI-mediated signaling pathway induces basophil activation.

[0384] 62. The method according to any one of paragraphs 1-61, wherein activation of the FcεRI-mediated signaling pathway induces mast cell activation.

[0385] 63. The method according to any one of paragraphs 1 to 62, wherein activation of the FcεRI-mediated signaling pathway induces basophil degranulation.

[0386] 64. The method according to any one of paragraphs 1-63, wherein activation of the FcεRI-mediated signaling pathway induces mast cell degranulation.

[0387] 65. A method comprising contacting a plasma sample or fraction thereof with a reagent that binds to anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in the plasma sample or fraction thereof, and detecting the binding of the reagent.

[0388] 66. The method according to paragraph 65, wherein the reagent directly binds to anti-FcεRI Ig in a plasma sample or fraction thereof.

[0389] 67. The method according to paragraph 65, wherein the reagent indirectly binds to anti-FcεRI Ig in a plasma sample or fraction thereof.

[0390] 68. The method according to paragraph 65 or 66, wherein the reagent comprises a ligand containing FcεRI or a fragment or epitope thereof.

[0391] 69. A method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, the method comprising determining the percentage of basophil and / or mast cell activation and / or degranulation induced by the plasma sample or fraction thereof, wherein the presence of basophil and / or mast cell activation and / or degranulation indicates the presence of anti-FcεRI Ig in the plasma sample or fraction thereof.

[0392] 70. The method according to any one of paragraphs 1-69, wherein the anti-FcεRI Ig is anti-FcεRI IgG and anti-FcεRI IgE.

[0393] 71. The method according to any one of paragraphs 1-70, wherein anti-FcεRI Ig is anti-FcεRI IgG.

[0394] 72. The method according to paragraph 70 or 71, wherein the anti-FcεRI IgG is IgG1.

[0395] 73. The method according to paragraph 70 or 71, wherein the anti-FcεRI IgG is IgG2.

[0396] 74. The method according to paragraph 70 or 71, wherein the anti-FcεRI IgG is IgG3.

[0397] 75. The method according to paragraph 70 or 71, wherein the anti-FcεRI IgG is IgG4.

[0398] 76. The method according to any one of paragraphs 1-69, wherein anti-FcεRI Ig is anti-FcεRI IgE.

[0399] 77. The method according to any one of paragraphs 1-69, wherein anti-FcεRI Ig is anti-FcεRI IgM.

[0400] 78. The method described in any one of paragraphs 1-69, wherein anti-FcεRI Ig is anti-FcεRI IgA.

[0401] 79. The method according to any one of paragraphs 1 to 78, wherein the ligand comprises FcεRI or a fragment or epitope thereof.

[0402] 80. The method according to any one of paragraphs 1 to 78, wherein the ligand is FcεRI or a fragment or epitope thereof.

[0403] 81. The method according to any one of paragraphs 1 to 78, wherein the ligand comprises an FcεRI antibody or a fragment thereof.

[0404] 82. The method according to any one of paragraphs 1 to 78, wherein the ligand comprises an FcεRI DNA aptamer.

[0405] 83. The method according to any one of paragraphs 1 to 82, wherein the detected protein is an anti-IgG antibody.

[0406] 84. The method according to paragraph 83, wherein the detected protein is an anti-IgG1 antibody.

[0407] 85. The method according to paragraph 83, wherein the detected protein is an anti-IgG2 antibody.

[0408] 86. The method according to paragraph 83, wherein the detected protein is an anti-IgG3 antibody.

[0409] 87. The method according to paragraph 84, wherein the detected protein is an anti-IgG4 antibody.

[0410] 88. The method according to any one of paragraphs 1 to 82, wherein the detected protein is an anti-IgE antibody.

[0411] 89. The method according to any one of paragraphs 1 to 82, wherein the detected protein is an anti-IgM antibody.

[0412] 90. The method according to any one of paragraphs 1 to 82, wherein the detected protein is an anti-IgA antibody.

[0413] 91. The method according to any one of paragraphs 1 to 90, wherein the detectable label is selected from the group consisting of radioactive labels, enzymes, fluorescent labels, luminescent labels, bioluminescent labels, magnetic labels, prosthetic groups, and contrast agents.

[0414] 92. The method described in any one of paragraphs 1 to 91, wherein the detectable label is a radioactive label.

[0415] 93. The method according to any one of paragraphs 1 to 91, wherein the detectable label is an enzyme.

[0416] 94. The method according to any one of paragraphs 1 to 91, wherein the detectable label is a fluorescent label.

[0417] 95. The method according to any one of paragraphs 1 to 91, wherein the detectable marker is a luminescent marker.

[0418] 96. The method according to any one of paragraphs 1 to 91, wherein the detectable label is a bioluminescent label.

[0419] 97. The method according to any one of paragraphs 1 to 91, wherein the detectable marker is a magnetic marker.

[0420] 98. The method according to any one of paragraphs 1 to 91, wherein the detectable label is a prosthetic group.

[0421] 99. The method according to any one of paragraphs 1 to 91, wherein the detectable label is a contrast agent.

[0422] 100. The method according to any one of paragraphs 1 to 99, further comprising determining the level of anti-FcεRI Ig in a plasma sample or fraction thereof.

[0423] 101. The method according to paragraph 98 or 99, further comprising administering a plasma sample or fraction thereof to a subject if the level of anti-FcεRI IgG is below a detectable level.

[0424] 102. The method according to paragraph 100 or 101, wherein anti-FcεRI Ig is anti-FcεRI IgG.

[0425] 103. The method according to paragraph 101 or 102, wherein the detectable amount of anti-FcεRI IgG in the plasma sample or fraction thereof is at least 40 μg / mL.

[0426] 104. The method according to paragraph 100, wherein anti-FcεRI Ig is anti-FcεRI IgE.

[0427] 105. The method according to paragraph 104, wherein the detectable amount of anti-FcεRI IgE in the plasma sample or fraction thereof is at least 0.1 kU / L.

[0428] 106. The method according to any one of paragraphs 1 to 105, further comprising determining the percentage of basophil activation induced by a plasma sample or fraction thereof.

[0429] 107. The method according to paragraph 106, wherein the rate of basophil activation after exposure to a plasma sample or fraction is determined by determining the percentage of basophils expressing one or more markers selected from the group consisting of CD63, CD203c, and combinations thereof, in the whole population of basophils exposed to the plasma sample or fraction.

[0430] 108. The method according to paragraph 106 or 107, wherein the rate of basophil activation after exposure to a plasma sample or fraction is determined by determining the percentage of CD63-expressing basophils in the entire population of basophils exposed to the plasma sample or fraction.

[0431] 109. The method according to paragraph 106 or 107, wherein the rate of basophil activation after exposure to a plasma sample or fraction is determined by determining the percentage of basophils expressing CD203c in the whole population of basophils exposed to the plasma sample or fraction.

[0432] 110. The method according to paragraph 106 or 107, wherein the percentage of basophil activation after exposure to a plasma sample or fraction is determined by determining the percentage of basophils expressing CD63 and CD203c in the whole population of basophils exposed to the plasma sample or fraction.

[0433] 111. The method according to any one of paragraphs 106-110, wherein the percentage of basophil activation induced by a plasma sample or fraction indicates the presence of a detectable amount of anti-FcεRI Ig in the plasma sample or fraction.

[0434] 112. The method according to paragraph 111, wherein a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction if the percentage of basophils expressing CD63 after exposure to the plasma sample or fraction is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of the total population of basophils exposed to the plasma sample or fraction.

[0435] 113. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 5% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0436] 114. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 6% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0437] 115. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 7% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0438] 116. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 8% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0439] 117. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 9% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0440] 118. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 10% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0441] 119. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 11% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0442] 120. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 12% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0443] 121. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 13% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0444] 122. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 14% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0445] 123. The method according to paragraph 111 or 112, wherein if the proportion of basophils expressing CD63 after exposure to a plasma sample or fraction exceeds 15% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0446] 124. The method according to paragraph 111 or 112, further comprising administering a plasma sample or fraction to a subject if anti-FcεRI Ig is not present in a detectable amount in the plasma sample or fraction.

[0447] 125. The method according to paragraph 124, wherein anti-FcεRI Ig is not present in detectable amounts if the percentage of basophils expressing CD63 after exposure to a plasma sample or fraction is less than 5% of the total population of basophils exposed to the plasma sample or fraction.

[0448] 126. The method according to paragraph 111, wherein a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction if the percentage of basophils expressing CD203c after exposure to the plasma sample or fraction is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of the total population of basophils exposed to the plasma sample or fraction.

[0449] 127. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 5% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0450] 128. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 6% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0451] 129. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 7% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0452] 130. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 8% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0453] 131. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 9% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0454] 132. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 10% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0455] 133. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 11% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0456] 134. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 12% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0457] 135. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 13% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0458] 136. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 14% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0459] 137. The method according to paragraph 111 or 126, wherein if the proportion of basophils expressing CD203c after exposure to a plasma sample or fraction exceeds 15% of the total population of basophils exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0460] 138. The method according to paragraph 111 or 124, wherein anti-FcεRI Ig is not present in detectable amounts if the percentage of basophils expressing CD203c after exposure to a plasma sample or fraction thereof is less than 5%.

[0461] 139. The method according to any one of paragraphs 106-138, wherein the percentage of basophil activation is determined by incubating basophils with a plasma sample or fraction thereof and quantifying the percentage of CD63-expressing basophils and / or CD203c-expressing basophils in the whole population of basophils using flow cytometry.

[0462] 141. The method according to any one of paragraphs 1 to 139, further comprising determining the percentage of mast cell activation induced by a plasma sample or fraction thereof.

[0463] 142. The method according to paragraph 141, wherein the rate of mast cell activation after exposure to a plasma sample or fraction is determined by determining the percentage of mast cells expressing CD107a in the entire population of mast cells exposed to the plasma sample or fraction.

[0464] 143. The method according to paragraph 141 or 142, wherein the percentage of mast cell activation induced by a plasma sample or fraction indicates the presence of a detectable amount of anti-FcεRI Ig in the plasma sample or fraction.

[0465] 144. The method according to paragraph 143, wherein a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction if the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 11%, greater than 12%, greater than 13%, greater than 14%, or greater than 15% of the total population of mast cells exposed to the plasma sample or fraction.

[0466] 145. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 5% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0467] 146. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 6% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0468] 147. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 7% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0469] 148. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 8% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0470] 149. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 9% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0471] 150. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 10% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0472] 151. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to a plasma sample or fraction exceeds 11% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0473] 152. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to a plasma sample or fraction exceeds 12% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0474] 153. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction exceeds 13% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0475] 154. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to a plasma sample or fraction exceeds 14% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0476] 155. The method according to paragraph 143 or 144, wherein if the proportion of mast cells expressing CD107a after exposure to a plasma sample or fraction exceeds 15% of the total population of mast cells exposed to the plasma sample or fraction, a detectable amount of anti-FcεRI Ig is present in the plasma sample or fraction.

[0477] 156. The method according to paragraph 143 or 144, wherein anti-FcεRI Ig is not present in detectable amounts in the plasma sample or fraction if the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction is less than 5%.

[0478] 157. The method according to any one of paragraphs 143-156, wherein the rate 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 whole population of mast cells using flow cytometry.

[0479] 158. The method according to any one of paragraphs 1 to 157, wherein the plasma sample or fraction thereof is selected from the group consisting of human blood plasma sample, IgG intermediate product, intravenous immunoglobulin G (IVIG), subcutaneous immunoglobulin G (SCIG), cryo-rich plasma, decryoplasma, supernatant I (SN I), Cohn fraction II (Fr II), Cohn fraction II+III (Fr II+III), Cohn fraction I+II+III (Fr I+II+III), Kistler / Nitzchemann precipitate A (KN A), Kistler / Nitzchemann precipitate B (KN B), Kistler / Nitzchemann precipitate of supernatant B (KN B+1), and combinations thereof.

[0480] 159. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is a human blood plasma sample.

[0481] 160. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is an IgG intermediate product.

[0482] 161. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is intravenous immunoglobulin G (IVIG).

[0483] 162. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is subcutaneous immunoglobulin G (SCIG).

[0484] 163. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is decryoplasma.

[0485] 164. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is supernatant I (SN I).

[0486] 165. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is Cohn fraction II (Fr II).

[0487] 166. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is Cohn fraction II+III (Fr II+III).

[0488] 167. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is Cohn fraction I+II+III (FrI+II+III).

[0489] 168. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is Kistler / Nitzschemann precipitate A (KN A).

[0490] 169. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is Kistler / Nitzschemann precipitate B (KN B).

[0491] 170. The method according to any one of paragraphs 1 to 158, wherein the plasma sample or fraction thereof is the Kistler / Nitschmann precipitate (KN B+1) of the supernatant B.

[0492] 171. The method according to any one of paragraphs 1 to 170, wherein the plasma sample or fraction thereof is a human blood plasma sample from one or more human subjects.

[0493] 172. An affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig immobilized in the matrix of the affinity chromatography resin.

[0494] 173. The affinity chromatography resin according to paragraph 172, further comprising blood type A antigens and blood type B antigens immobilized on a matrix of affinity chromatography resin.

[0495] 174. A method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation from a plasma sample or fraction thereof, the method comprising: conjugating anti-FcεRI Ig to an affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig in a plasma sample or fraction thereof; and collecting the anti-FcεRI Ig depletion preparation.

[0496] 175. The method according to paragraph 174, further comprising loading a plasma sample or fraction thereof onto an affinity chromatography resin.

[0497] 176. A method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation from a plasma sample or fraction thereof, wherein the method is: (i) Loading a plasma sample or fraction into an affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig in the plasma sample or fraction, which is immobilized on the matrix of the affinity chromatography resin, (ii) A method comprising collecting an anti-FcεRI Ig reduction preparation.

[0498] 177. The method according to any one of paragraphs 174-176, wherein the method further comprises a washing and / or elution step.

[0499] 178. The method according to any one of paragraphs 174-177, wherein the method further comprises an elution step.

[0500] 179. The method according to any one of paragraphs 174-178, wherein the method further includes a washing step.

[0501] 180. The method according to any one of paragraphs 174-179, wherein anti-FcεRI Ig binds to the α-chain of FcεRI and activates the FcεRI-mediated signaling pathway.

[0502] 181. The method according to any one of paragraphs 174-180, wherein an anti-FcεRI Ig-reduced preparation induces reduced activation of the FcεRI-mediated signaling pathway compared to a preparation in which anti-FcεRI Ig is not reduced.

[0503] 182. The method according to any one of paragraphs 174-181, wherein anti-FcεRI Ig is not present in a detectable amount in the anti-FcεRI Ig-reduced preparation.

[0504] 183. The method according to paragraph 182, wherein an anti-FcεRI Ig-reduced preparation induces reduced activation of the FcεRI-mediated signaling pathway compared to a preparation in which anti-FcεRI Ig is not reduced.

[0505] 184. The method according to any one of paragraphs 174-182, wherein anti-FcεRI Ig is not present in a detectable amount, and the anti-FcεRI Ig-reduced preparation induces reduced basophil activation compared to a preparation in which anti-FcεRI Ig is not reduced.

[0506] 185. The method according to any one of paragraphs 174-182, wherein anti-FcεRI Ig is not present in a detectable amount, and the anti-FcεRI Ig-reduced preparation induces reduced mast cell activation compared to a preparation in which anti-FcεRI Ig is not reduced.

[0507] 186. The method according to any one of paragraphs 174-184, wherein basophil activation after exposure to an anti-FcεRI Ig-reducing preparation is determined by determining the percentage of basophils expressing one or more markers selected from the group consisting of CD63, CD203c, and combinations thereof.

[0508] 187. The method according to paragraph 186, wherein basophil activation is determined by determining the percentage of basophils expressing CD63 after exposure to an anti-FcεRI Ig-reduced preparation.

[0509] 188. The method according to paragraph 186, wherein basophil activation is determined by determining the percentage of basophils CD203c after exposure to an anti-FcεRI Ig reduced preparation.

[0510] 189. The method according to paragraph 186, wherein basophil activation is determined by determining the percentage of basophils expressing CD63 and CD203c after exposure to an anti-FcεRI Ig-reduced preparation.

[0511] 190. The method according to any one of paragraphs 186, 187, or 189, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation 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 anti-FcεRI Ig-reducing preparation.

[0512] 191. The method according to any one of paragraphs 186, 187, 189, or 190, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0513] 192. The method according to any one of paragraphs 186, 187, 189-191, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 4% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0514] 193. The method according to any one of paragraphs 186, 187, 189-192, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 3% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0515] 194. The method according to any one of paragraphs 186, 187, 189-193, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 2% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0516] 195. The method according to any one of paragraphs 186, 187, 189-194, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 1% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0517] 196. The method according to any one of paragraphs 186, 187, or 189, further comprising administering an anti-FcεRI Ig-reducing preparation to a subject if the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0518] 197. The method or use according to any one of paragraphs 186, 187, or 189, further comprising including an anti-FcεRI Ig depletion preparation in a pooled blood plasma sample if the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig depletion preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig depletion preparation.

[0519] 198. The method according to any one of paragraphs 186, 187, or 189, further comprising excluding the anti-FcεRI Ig depletion preparation from a pooled blood plasma sample if the method or use further comprises excluding the anti-FcεRI Ig depletion preparation from a pooled blood plasma sample if the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig depletion preparation exceeds 5% of the total population of basophils exposed to the anti-FcεRI Ig depletion preparation.

[0520] 199. The method according to any one of paragraphs 186, 187, or 189, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation 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 anti-FcεRI Ig-reducing preparation.

[0521] 200. The method according to any one of paragraphs 186, 187, 189, or 199, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0522] 201. The method according to any one of paragraphs 186, 187, 189, 199, or 200, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 4% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0523] 202. The method according to any one of paragraphs 186, 187, 189, 199-201, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 3% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0524] 203. The method according to any one of paragraphs 186, 187, 189, and 199-202, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 2% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0525] 204. The method according to any one of paragraphs 186, 187, 189, and 199-203, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 1% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0526] 205. The method according to any one of paragraphs 186, 187, or 189, further comprising administering an anti-FcεRI Ig-reducing preparation to a subject if the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0527] 206. The method according to any one of paragraphs 186, 187, or 189, further comprising including an anti-FcεRI Ig-reducing preparation in a pooled blood plasma sample if the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation.

[0528] 207. The method according to any one of paragraphs 186, 187, or 189, further comprising excluding an anti-FcεRI Ig-depleting preparation from a pooled blood plasma sample if the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-depleting preparation exceeds 5% of the total population of basophils exposed to the anti-FcεRI Ig-depleting preparation.

[0529] 208. The method according to any one of paragraphs 185-207, wherein mast cell activation after exposure to an anti-FcεRI Ig-reducing preparation is determined by determining the percentage of mast cells expressing CD107a.

[0530] 209. The method according to paragraph 208, wherein the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation 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 anti-FcεRI Ig-reducing preparation.

[0531] 210. The method according to paragraph 208 or 210, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0532] 211. The method according to paragraphs 208-210, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 4% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0533] 212. The method according to paragraphs 208-211, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 3% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0534] 213. The method according to paragraphs 208-212, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 2% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0535] 214. The method according to paragraphs 208-213, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 1% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0536] 215. The method according to paragraphs 208-214, further comprising administering an anti-FcεRI Ig-reducing preparation to subjects if the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0537] 216. The method according to paragraphs 208-214, further comprising including an anti-FcεRI Ig-reducing preparation in a pooled blood plasma sample if the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0538] 217. The method according to paragraphs 208-214, further comprising excluding the anti-FcεRI Ig-reducing preparation from a pooled blood plasma sample if the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation exceeds 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

[0539] 218. The method according to any one of paragraphs 186-198, wherein the percentage of basophil activation is determined by incubating basophils with an anti-FcεRI Ig depleted preparation and quantifying the percentage of CD63-expressing basophils in the whole basophil population using flow cytometry.

[0540] 219. The method according to any one of paragraphs 199-207, wherein the percentage of basophil activation is determined by incubating basophils with an anti-FcεRI Ig depletion preparation and quantifying the percentage of CD203c-expressing basophils in the whole basophil population using flow cytometry.

[0541] 220. The method according to any one of paragraphs 208-217, wherein the rate of mast cell activation is determined by incubating mast cells with an anti-FcεRI Ig-reduced preparation and quantifying the percentage of CD107a-expressing mast cells in the whole population of mast cells using flow cytometry.

[0542] 221. The method according to any one of paragraphs 174-220, wherein the anti-FcεRI Ig reduction preparation contains less than 40 μg / mL of anti-FcεRI IgG.

[0543] 222. The method according to any one of paragraphs 174-221, wherein the anti-FcεRI Ig reduction preparation contains less than 35 μg / mL of anti-FcεRI IgG.

[0544] 223. The method according to any one of paragraphs 174-222, wherein the anti-FcεRI Ig reduction preparation contains less than 30 μg / mL of anti-FcεRI IgG.

[0545] 224. The method according to any one of paragraphs 174-223, wherein the anti-FcεRI Ig reduction preparation contains anti-FcεRI IgG less than 25 μg / mL.

[0546] 225. The method according to any one of paragraphs 174-224, wherein the anti-FcεRI Ig reduction preparation contains less than 20 μg / mL of anti-FcεRI IgG.

[0547] 226. The method according to any one of paragraphs 174-225, wherein the anti-FcεRI Ig reduction preparation contains less than 15 μg / mL of anti-FcεRI IgG.

[0548] 227. The method according to any one of paragraphs 174-226, wherein the anti-FcεRI Ig reduction preparation contains less than 10 μg / mL of anti-FcεRI IgG.

[0549] 228. The method according to any one of paragraphs 174-227, wherein the anti-FcεRI Ig reduction preparation contains less than 5 μg / mL of anti-FcεRI IgG.

[0550] 229. The method according to any one of paragraphs 174-228, wherein the anti-FcεRI Ig reduction preparation contains 0-39.9 μg / mL of anti-FcεRI IgG.

[0551] 230. The method according to any one of paragraphs 174-229, wherein the anti-FcεRI Ig reduction preparation contains 0-30 μg / mL of anti-FcεRI IgG.

[0552] 231. The method according to any one of paragraphs 174-230, wherein the anti-FcεRI Ig reduction preparation contains 0-20 μg / mL of anti-FcεRI IgG.

[0553] 232. The method according to any one of paragraphs 174-231, wherein the anti-FcεRI Ig reduction preparation contains 0-10 μg / mL of anti-FcεRI IgG.

[0554] 233. The method according to any one of paragraphs 174-232, wherein the anti-FcεRI Ig depleting preparation contains less than 0.1 kU / L of anti-FcεRI IgE.

[0555] 234. The method according to any one of paragraphs 174-233, wherein the anti-FcεRI Ig depleting preparation contains less than 0.05 kU / L of anti-FcεRI IgE.

[0556] 235. The method according to any one of paragraphs 174-234, wherein the anti-FcεRI Ig depleting preparation contains less than 0.01 kU / L of anti-FcεRI IgE.

[0557] 236. The method according to any one of paragraphs 174-235, wherein the anti-FcεRI Ig depleting preparation contains less than 0.001 kU / L of anti-FcεRI IgE.

[0558] 237. The method according to any one of paragraphs 174-236, wherein the anti-FcεRI Ig reduction preparation contains 0 kU / L anti-FcεRI IgE.

[0559] 238. The method according to any one of paragraphs 174-237, wherein the anti-FcεRI Ig in the preparation is reduced to 0-0.1 kU / L anti-FcεRI IgE.

[0560] 239. The method according to any one of paragraphs 174-238, wherein the anti-FcεRI Ig concentration in the preparation is reduced to 0.05-0.1 kU / L anti-FcεRI IgE.

[0561] 240. The method according to any one of paragraphs 174-239, wherein the affinity chromatography resin further comprises blood group A antigen immobilized on a matrix of affinity chromatography resin.

[0562] 241. The method according to any one of paragraphs 174 to 240, wherein the affinity chromatography resin is a continuous affinity chromatography resin.

[0563] 242. Affinity chromatography, (i) A first affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the first affinity chromatography resin, (ii) The method according to paragraph 240 or 241, comprising a second affinity chromatography resin containing a blood group A antigen immobilized on a matrix of the second affinity chromatography resin, and a continuous affinity chromatography method.

[0564] 243. Affinity chromatography, (i) A first affinity chromatography resin containing blood group A antigen immobilized on the matrix of a second affinity chromatography resin, (ii) The method according to paragraph 240 or 241, comprising a second affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, immobilized on a matrix of a first affinity chromatography resin, and being a continuous affinity chromatography.

[0565] 244. The method according to any one of paragraphs 174-243, wherein the affinity chromatography resin further comprises blood group B antigen immobilized on the matrix of the affinity chromatography resin.

[0566] 245. Affinity chromatography, (i) A first affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the first affinity chromatography resin, (ii) The method according to paragraph 244, comprising a second affinity chromatography resin containing a blood group B antigen immobilized on a matrix of the second affinity chromatography resin, and a continuous affinity chromatography method.

[0567] 246. Affinity chromatography, (i) A first affinity chromatography resin containing blood group B antigen immobilized on the matrix of a second affinity chromatography resin, (ii) The method according to paragraph 244, comprising a second affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, immobilized on a matrix of a first affinity chromatography resin, and being a continuous affinity chromatography.

[0568] 247. The method according to any one of paragraphs 174 to 239, wherein the affinity chromatography resin further comprises blood group A antigens and blood group B antigens immobilized on a matrix of the affinity chromatography resin.

[0569] 248. Affinity chromatography, (i) A first affinity chromatography resin comprising a ligand that binds to anti-FcεRI Ig, immobilized on the matrix of the first affinity chromatography resin, (ii) The method according to paragraph 247, comprising a second affinity chromatography resin containing blood group A antigen and blood group B antigen immobilized on a matrix of the second affinity chromatography resin, wherein the method is a continuous affinity chromatography.

[0570] 249. Affinity chromatography, (i) A first affinity chromatography resin containing blood type A antigen and blood type B antigen immobilized on a matrix of a second affinity chromatography resin, (ii) The method according to paragraph 247, comprising a second affinity chromatography resin containing a ligand that binds to anti-FcεRI Ig, immobilized on a matrix of a first affinity chromatography resin, wherein the method is a continuous affinity chromatography.

[0571] 250. The method according to any one of paragraphs 1 to 249, wherein the ligand that binds to anti-FcεRI Ig comprises FcεRI or a fragment or epitope thereof.

[0572] 251. The method according to any one of paragraphs 1 to 250, wherein the method further comprises one or more steps selected from the group consisting of ethanol precipitation, octanoid acid fractionation, ion exchange chromatography, virus inactivation, virus filtration, ultrafiltration / dialysis filtration, and combinations thereof.

[0573] 252. The method according to paragraph 251, further comprising the step of ethanol precipitation.

[0574] 253. The method according to paragraph 251 or 252, wherein the method further comprises the step of octanoid acid fractionation.

[0575] 254. The method according to any one of paragraphs 251-253, wherein the method further comprises a step of ion exchange chromatography.

[0576] 255. The method according to any one of paragraphs 251-254, wherein the method further includes a step of virus inactivation.

[0577] 256. The method according to any one of paragraphs 251-255, wherein the method further includes a step of virus filtration.

[0578] 257. The method according to any one of paragraphs 1 to 256, wherein the method further includes the step of ultrafiltration / diafiltration.

[0579] 258. Anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion IgG preparation containing polyclonal IgG.

[0580] 259. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 258, wherein 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), Xembify® (Grifols), Cutaquig® (Octapharma), and Cuvitru® (Takeda).

[0581] 260. The anti-FcεRI Ig-reducing IgG preparation according to paragraph 258 or 259, wherein the preparation further comprises a stabilizer containing proline and has a pH of about 4.2 to about 5.4.

[0582] 261. The anti-FcεRI Ig-decreasing IgG preparation described in paragraph 260, in which proline is present at a concentration of 250 mM and the pH is 4.8.

[0583] An anti-FcεRI Ig-decreasing IgG preparation as described in any one of paragraphs 258-261, further comprising 262.20 μg / mL of PS80, with proline present at a concentration of 250 mM and a pH of 4.8.

[0584] 263. An anti-FcεRI Ig-reduced IgG preparation according to any one of paragraphs 258-262, wherein the anti-FcεRI Ig-reduced IgG preparation does not contain a detectable amount of anti-FcεRI Ig and induces reduced activation of the FcεRI-mediated signaling pathway compared to an IgG preparation in which anti-FcεRI Ig is not reduced.

[0585] 264. An anti-FcεRI Ig-reduced IgG preparation according to any one of paragraphs 258-263, wherein the anti-FcεRI Ig-reduced IgG preparation does not contain a detectable amount of anti-FcεRI Ig and induces reduced basophil activation compared to a preparation in which anti-FcεRI Ig is not reduced.

[0586] 265. An anti-FcεRI Ig-reduced IgG preparation according to any one of paragraphs 258-264, wherein the anti-FcεRI Ig-reduced IgG preparation does not contain a detectable amount of anti-FcεRI Ig and induces reduced mast cell activation compared to a preparation in which anti-FcεRI Ig is not reduced.

[0587] 266. An anti-FcεRI Ig-reducing IgG preparation according to any one of paragraphs 258 to 265, wherein the anti-FcεRI Ig-reducing IgG preparation contains anti-FcεRI IgG in a concentration of less than 40 μg / mL, less than 35 μg / mL, less than 30 μg / mL, less than 25 μg / mL, less than 20 μg / mL, less than 15 μg / mL, less than 10 μg / mL, or less than 5 μg / mL.

[0588] 267. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 40 μg / mL of anti-FcεRI IgG.

[0589] 268. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 35 μg / mL of anti-FcεRI IgG.

[0590] 269. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 30 μg / mL of anti-FcεRI IgG.

[0591] 270. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 25 μg / mL of anti-FcεRI IgG.

[0592] 271. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 20 μg / mL of anti-FcεRI IgG.

[0593] 272. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 15 μg / mL of anti-FcεRI IgG.

[0594] 273. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 10 μg / mL of anti-FcεRI IgG.

[0595] 274. The anti-FcεRI Ig-reducing IgG preparation according to paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 5 μg / mL of anti-FcεRI IgG.

[0596] 275. The anti-FcεRI Ig-reducing IgG preparation according to paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 1 μg / mL of anti-FcεRI IgG.

[0597] 276. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 40 μg / mL of anti-FcεRI IgG.

[0598] 277. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 35 μg / mL of anti-FcεRI IgG.

[0599] 278. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 30 μg / mL of anti-FcεRI IgG.

[0600] 279. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 25 μg / mL of anti-FcεRI IgG.

[0601] 280. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 20 μg / mL of anti-FcεRI IgG.

[0602] 281. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 15 μg / mL of anti-FcεRI IgG.

[0603] 282. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 10 μg / mL of anti-FcεRI IgG.

[0604] 283. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 266, wherein the anti-FcεRI Ig-reducing IgG preparation contains 1 to 5 μg / mL of anti-FcεRI IgG.

[0605] 284. An anti-FcεRI Ig-reducing IgG preparation according to any one of paragraphs 258 to 283, wherein the anti-FcεRI Ig-reducing IgG preparation contains anti-FcεRI IgE in a concentration of less than 0.1 kU / L, less than 0.05 kU / L, less than 0.01 kU / L, or less than 0.001 kU / L.

[0606] 285. The anti-FcεRI Ig-decreasing IgG preparation according to paragraph 284, wherein the anti-FcεRI Ig-decreasing IgG preparation contains less than 0.1 kU / L of anti-FcεRI IgE.

[0607] 286. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 284, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 0.05 kU / L of anti-FcεRI IgE.

[0608] 287. The anti-FcεRI Ig-reducing IgG preparation according to paragraph 284, wherein the anti-FcεRI Ig-reducing IgG preparation contains less than 0.01 kU / L of anti-FcεRI IgE.

[0609] 288. The anti-FcεRI Ig-decreasing IgG preparation according to paragraph 284, wherein the anti-FcεRI Ig-decreasing IgG preparation contains less than 0.001 kU / L of anti-FcεRI IgE.

[0610] 289. The anti-FcεRI Ig-decreasing IgG preparation described in paragraph 284, wherein the anti-FcεRI Ig-decreasing IgG preparation contains 0.001 to 0.1 kU / L of anti-FcεRI IgE.

[0611] 290. The anti-FcεRI Ig-decreasing IgG preparation described in paragraph 284, wherein the anti-FcεRI Ig-decreasing IgG preparation contains 0.001 to 0.05 kU / L of anti-FcεRI IgE.

[0612] 291. The anti-FcεRI Ig-decreasing IgG preparation described in paragraph 284, wherein the anti-FcεRI Ig-decreasing IgG preparation contains 0.001 to 0.01 kU / L of anti-FcεRI IgE.

[0613] 292. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 258-291, wherein basophil activation after exposure to the anti-FcεRI Ig-decreasing IgG preparation is determined by determining the percentage of basophils expressing one or more markers selected from the group consisting of CD63, CD203c, and combinations thereof.

[0614] 293. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 258-292, wherein basophil activation after exposure to the anti-FcεRI Ig-decreasing IgG preparation is determined by determining the percentage of basophils expressing CD63.

[0615] 294. An anti-FcεRI Ig-decreased IgG preparation as described in any one of paragraphs 258-293, wherein basophil activation after exposure to the anti-FcεRI Ig-decreased IgG preparation is determined by determining the percentage of basophils expressing CD203c.

[0616] 295. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 258-294, wherein basophil activation after exposure to the anti-FcεRI Ig-decreasing IgG preparation is determined by determining the percentage of basophils expressing CD63 and CD203c.

[0617] 296. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 293, and 295, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-decreasing IgG preparation 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 anti-FcεRI Ig-decreasing IgG preparation.

[0618] 297. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 293, 295, or 296, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 1% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0619] 298. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 293, 295, or 296, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 2% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0620] 299. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 293, 295, or 296, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 3% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0621] 300. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 293, 295, or 296, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 4% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0622] 301. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 293, 295, or 296, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0623] 302. An anti-FcεRI Ig-decreased IgG preparation as described in any one of paragraphs 292, 293, or 295-301, wherein the proportion of basophil activation is determined by incubating basophils with an anti-FcεRI Ig-decreased IgG preparation and quantifying the proportion of CD63-expressing basophils in the whole population of basophils using flow cytometry.

[0624] 303. An anti-FcεRI Ig-reduced IgG preparation according to any one of paragraphs 292, 294, or 295, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reduced IgG preparation 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 anti-FcεRI Ig-reduced IgG preparation.

[0625] 304. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 294, 295, or 303, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 1% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0626] 305. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 294, 295, or 303, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 2% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0627] 306. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 294, 295, or 303, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 3% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0628] 307. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 294, 295, or 303, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 4% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0629] 308. An anti-FcεRI Ig-decreasing IgG preparation according to any one of paragraphs 292, 294, 295, or 303, wherein the proportion of basophils expressing CD203c after exposure to the anti-FcεRI Ig-decreasing IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-decreasing IgG preparation.

[0630] 309. An anti-FcεRI Ig-decreased IgG preparation according to any one of paragraphs 292, 294, 295, or 303, wherein the proportion of basophil activation is determined by incubating basophils with an anti-FcεRI Ig-decreased IgG preparation and quantifying the proportion of CD203c-expressing basophils in the whole population of basophils using flow cytometry.

[0631] 310. An anti-FcεRI Ig-reduced IgG preparation as described in any one of paragraphs 258-291, wherein mast cell activation after exposure to the anti-FcεRI Ig-reduced IgG preparation is determined by determining the percentage of mast cells expressing CD107a.

[0632] 311. The anti-FcεRI Ig-reducing IgG preparation described in paragraph 310, wherein the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing IgG preparation 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 anti-FcεRI Ig-reducing IgG preparation.

[0633] 312. An anti-FcεRI Ig-reducing IgG preparation according to paragraph 310 or 311, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing IgG preparation is less than 1% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing IgG preparation.

[0634] 313. An anti-FcεRI Ig-reducing IgG preparation according to paragraph 310 or 311, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing IgG preparation is less than 2% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing IgG preparation.

[0635] 314. An anti-FcεRI Ig-reducing IgG preparation according to paragraph 310 or 311, wherein the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing IgG preparation is less than 3% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing IgG preparation.

[0636] 315. An anti-FcεRI Ig-reducing IgG preparation according to paragraph 310 or 311, wherein the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing IgG preparation is less than 4% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing IgG preparation.

[0637] 316. An anti-FcεRI Ig-reducing IgG preparation according to paragraph 310 or 311, wherein the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing IgG preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing IgG preparation.

[0638] 317. An anti-FcεRI Ig-reducing IgG preparation as described in any one of paragraphs 310-316, wherein the rate of mast cell activation is determined by incubating mast cells with an anti-FcεRI Ig-reducing IgG preparation and quantifying the percentage of CD107a-expressing mast cells in the whole population of mast cells using flow cytometry.

[0639] 318. A pharmaceutical composition comprising an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depletion preparation prepared by any one of paragraphs 174 to 257.

[0640] 319. A pharmaceutical composition comprising an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig)-reducing IgG preparation described in any one of paragraphs 258 to 317.

[0641] 320. A pooled IgG preparation comprising a detectable amount of anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig).

[0642] 321. The pooled IgG preparation according to paragraph 320, comprising a detectable amount of anti-FcεRI Ig greater than 40 μg / mL, greater than 45 μg / mL, greater than 50 μg / mL, greater than 55 μg / mL, or greater than 60 μg / mL of anti-FcεRI IgG.

[0643] 322. A pooled IgG preparation according to either paragraph 320 or 321, comprising anti-FcεRI IgG in a detectable amount of anti-FcεRI Ig greater than 40 μg / mL.

[0644] 323. A pooled IgG preparation according to any one of paragraphs 320-322, comprising anti-FcεRI IgG in a detectable amount of anti-FcεRI Ig greater than 45 μg / mL.

[0645] 324. A pooled IgG preparation according to any one of paragraphs 320-323, comprising anti-FcεRI IgG in a detectable amount of anti-FcεRI Ig greater than 50 μg / mL.

[0646] 325. A pooled IgG preparation according to any one of paragraphs 320-324, comprising anti-FcεRI IgG in a detectable amount of anti-FcεRI Ig greater than 55 μg / mL.

[0647] 326. A pooled IgG preparation according to any one of paragraphs 320-325, comprising anti-FcεRI IgG in a detectable amount greater than 60 μg / mL of anti-FcεRI Ig.

[0648] 327. A pooled IgG preparation according to any one of paragraphs 320-326, comprising a detectable amount of anti-FcεRI Ig greater than 0.1 kU / L, greater than 0.5 kU / L, greater than 1 kU / L, or greater than 1.5 kU / L of anti-FcεRI IgE.

[0649] 328. A pooled IgG preparation according to any one of paragraphs 320-327, comprising anti-FcεRI IgE in a detectable amount greater than 0.1 kU / L of anti-FcεRI Ig.

[0650] 329. A pooled IgG preparation according to any one of paragraphs 320-328, comprising anti-FcεRI IgE in a detectable amount greater than 0.5 kU / L of anti-FcεRI Ig.

[0651] 330. A pooled IgG preparation according to any one of paragraphs 320-329, comprising anti-FcεRI IgE in a detectable amount greater than 1 kU / L of anti-FcεRI Ig.

[0652] 331. A pooled IgG preparation according to any one of paragraphs 320-330, comprising anti-FcεRI IgE in a detectable amount greater than 1.5 kU / L of anti-FcεRI Ig.

[0653] 332. A pooled IgG preparation according to any one of paragraphs 320-331, comprising a detectable amount of anti-FcεRI Ig containing 0.1-5 kU / L of anti-FcεRI IgE.

[0654] 333. A pooled IgG preparation according to any one of paragraphs 320-332, wherein basophil activation after exposure to the pooled IgG preparation is determined by determining the percentage of basophils expressing one or more markers selected from the group consisting of CD63, CD203c, and combinations thereof.

[0655] 334. A pooled IgG preparation according to any one of paragraphs 320-333, wherein basophil activation after exposure to the pooled IgG preparation is determined by determining the percentage of basophils expressing CD63.

[0656] 335. A pooled IgG preparation according to any one of paragraphs 320-333, wherein basophil activation after exposure to the pooled IgG preparation is determined by determining the percentage of basophils expressing CD203c.

[0657] 336. A pooled IgG preparation according to any one of paragraphs 320-333, wherein basophil activation after exposure to the pooled IgG preparation is determined by determining the percentage of basophils expressing CD63 and CD203c.

[0658] 337. A pooled IgG preparation according to any one of paragraphs 320-334 or 336, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, or greater than 10% of the total population of basophils exposed to the pooled IgG preparation.

[0659] 338. A pooled IgG preparation according to any one of paragraphs 320-334 or 336-337, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation is greater than 5% of the total population of basophils exposed to the pooled IgG preparation.

[0660] 339. A pooled IgG preparation according to any one of paragraphs 320-334 or 336-338, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation exceeds 6% of the total population of basophils exposed to the pooled IgG preparation.

[0661] 340. A pooled IgG preparation according to any one of paragraphs 320-334 or 336-339, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation exceeds 7% of the total population of basophils exposed to the pooled IgG preparation.

[0662] 341. A pooled IgG preparation according to either paragraph 320-334 or 336-340, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation exceeds 8% of the total population of basophils exposed to the pooled IgG preparation.

[0663] 342. A pooled IgG preparation according to any one of paragraphs 320-334 or 336-341, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation exceeds 9% of the total population of basophils exposed to the pooled IgG preparation.

[0664] 343. A pooled IgG preparation according to either paragraph 320-334 or 336-342, wherein the proportion of basophils expressing CD63 after exposure to the pooled IgG preparation is greater than 10% of the total population of basophils exposed to the pooled IgG preparation.

[0665] 344. The method according to either paragraph 320-334 or 336-342, wherein the percentage of basophil activation is determined by incubating basophils with a plasma sample or fraction thereof and quantifying the percentage of CD63-expressing basophils in the whole population of basophils using flow cytometry.

[0666] 345. A pooled IgG preparation according to any one of paragraphs 320-333 or 335, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, or greater than 10% of the total population of basophils exposed to the pooled IgG preparation.

[0667] 346. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation is greater than 5% of the total population of basophils exposed to the pooled IgG preparation.

[0668] 347. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345-346, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation is greater than 6% of the total population of basophils exposed to the pooled IgG preparation.

[0669] 348. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345-347, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation exceeds 7% of the total population of basophils exposed to the pooled IgG preparation.

[0670] 349. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345-348, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation exceeds 8% of the total population of basophils exposed to the pooled IgG preparation.

[0671] 350. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345-349, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation exceeds 9% of the total population of basophils exposed to the pooled IgG preparation.

[0672] 351. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345-350, wherein the proportion of basophils expressing CD203c after exposure to the pooled IgG preparation is greater than 10% of the total population of basophils exposed to the pooled IgG preparation.

[0673] 352. A pooled IgG preparation according to any one of paragraphs 320-333, 335, or 345-351, wherein the proportion of basophil activation is determined by incubating basophils with a pooled IgG preparation and quantifying the proportion of CD203c-expressing basophils in the total population of basophils using flow cytometry.

[0674] 353. A pooled IgG preparation according to any one of paragraphs 320-352, wherein mast cell activation after exposure to the pooled IgG preparation is determined by determining the proportion of mast cells expressing CD107a in the entire population of mast cells exposed to the pooled IgG preparation.

[0675] 354. A pooled IgG preparation according to paragraph 353, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, or greater than 10% of the total population of mast cells exposed to the pooled IgG preparation.

[0676] 355. A pooled IgG preparation according to paragraph 353 or 354, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 5% of the total population of mast cells exposed to the pooled IgG preparation.

[0677] 356. A pooled IgG preparation according to either paragraph 353 or 354, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 6% of the total population of mast cells exposed to the pooled IgG preparation.

[0678] 357. A pooled IgG preparation according to either paragraph 353 or 354, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 7% of the total population of mast cells exposed to the pooled IgG preparation.

[0679] 358. A pooled IgG preparation according to either paragraph 353 or 354, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 8% of the total population of mast cells exposed to the pooled IgG preparation.

[0680] 359. A pooled IgG preparation according to either paragraph 353 or 354, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 9% of the total population of mast cells exposed to the pooled IgG preparation.

[0681] 360. A pooled IgG preparation according to either paragraph 353 or 354, wherein the proportion of mast cells expressing CD107a after exposure to the pooled IgG preparation is greater than 10% of the total population of mast cells exposed to the pooled IgG preparation.

[0682] 361. A pooled IgG preparation according to any one of paragraphs 353-360, wherein the rate of mast cell activation is determined by incubating mast cells with the pooled IgG preparation and quantifying the proportion of CD107a-expressing mast cells in the entire population of mast cells using flow cytometry.

[0683] 362. A method for treating, preventing, and / or delaying the progression of a condition in a subject, comprising administering a pharmaceutical composition described in paragraph 318 or 319, or an anti-FcεRI Ig-reducing IgG preparation described in any one of paragraphs 320 to 361.

[0684] 363. A pharmaceutical composition as described in paragraph 318 or 319, or an anti-FcεRI Ig-reducing IgG preparation as described in any one of paragraphs 320 to 361, for use in treating, preventing, and / or delaying the progression of a condition in a subject.

[0685] 364. Use of a pharmaceutical composition described in paragraph 318 or 319, or an anti-FcεRI Ig-reducing IgG preparation described in any one of paragraphs 320 to 361, in the manufacture of a medicinal product for treating, preventing, and / or delaying the progression of a condition in a subject.

[0686] 365. Use of the method described in paragraph 362, or the pharmaceutical composition or anti-FcεRI Ig-reducing IgG preparation described in paragraph 363, or the use described in paragraph 364, for conditions related to immunodeficiency, inflammatory disease, autoimmune disease, or acute infection.

[0687] 366. Immunodeficiency, congenital agammaglobulinemia, congenital hypogammaglobulinemia, unclassified immunodeficiency, severe combined immunodeficiency, allogeneic bone marrow transplant, chronic lymphocytic leukemia, childhood HIV, kidney transplant with high antibody recipient or ABO-incompatible donor, chronic fatigue syndrome, Clostridium Difficile enteritis, 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, generalized rigidity syndrome, ocular clonus-myoclonus ataxia, severe sepsis and septic shock in critically ill adults, toxic epidermal necrolysis, chronic lymphocytic leukemia, multiple myeloma, X-linked agammaglobulinemia, hypogammaglobulinemia, RRMS, IgG subclass deficiency due to recurrent infection, and combinations thereof, selected from the group consisting of the following: the method or pharmaceutical composition or anti-FcεRI Ig-decreasing IgG preparation or use described in paragraph 365.

[0688] 367. Inflammatory or autoimmune diseases selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus (SLE), antiphospholipid syndrome, immune thrombocytopenia (ITP), Kawasaki disease, Guillain-Barré syndrome (GBS), multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CIDP), multifocal motor neuropathy (MMN), myasthenia gravis (MG), cutaneous bullous diseases, scleroderma, dermatomyositis, polymyositis, Alzheimer's disease, Parkinson's disease, Alzheimer's disease associated with Down syndrome, cerebral amyloid angiopathy, Lewy body dementia, frontotemporal dementia, vascular dementia, cell and organ transplantation, and combinations thereof, the method or pharmaceutical composition or anti-FcεRI Ig-reducing IgG preparation or use described in paragraph 365.

[0689] 368. A method for preparing an immunoglobulin (Ig) preparation from a plasma sample or fraction thereof, wherein the Ig preparation has a reduced adverse reaction in the subject, and the method comprises determining the rate of basophil and / or mast cell activation after exposure to the plasma sample or fraction thereof. (i) If the percentage of basophils expressing CD63 after exposure to the plasma sample or fraction 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, then include the plasma sample or fraction in the Ig preparation. (ii) If the percentage of basophils expressing CD63 after exposure to the plasma sample or fraction is greater than 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the total population of basophils exposed to the plasma sample or fraction, the plasma sample or fraction is excluded from the Ig preparation. (iii) If the percentage of basophils expressing CD203c after exposure to the plasma sample or fraction 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, then include the plasma sample or fraction in the Ig preparation. (iv) If the percentage of basophils expressing CD203c after exposure to the plasma sample or fraction is greater than 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the total population of basophils exposed to the plasma sample or fraction, the plasma sample or fraction is excluded from the Ig preparation. (v) If the percentage of mast cells expressing CD107a after exposure to the plasma sample or fraction 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, then include the plasma sample or fraction in the Ig preparation. (vi) If the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction is greater than 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% of the total population of mast cells exposed to the plasma sample or fraction, the plasma sample or fraction is excluded from the Ig preparation and / or (v) A method in which, if the plasma sample or fraction contains less than a detectable amount of Ig that causes an adverse reaction, the plasma sample or fraction is included in the Ig preparation, wherein the detectable amount of Ig that causes an adverse reaction is less than 40 μg / mL, less than 35 μg / mL, less than 30 μg / mL, less than 25 μg / mL, less than 20 μg / mL, less than 15 μg / mL, less than 10 μg / mL, or less than 5 μg / mL.

[0690] 369. The method described in paragraph 368, wherein the adverse reaction is a hypersensitivity reaction (HSR).

[0691] 370. The method according to paragraph 369, wherein HSR is selected from the group consisting of pruritus, erythema, urticaria, swelling, and combinations thereof.

[0692] 371. The method described in any one of paragraphs 368-370, wherein an adverse reaction occurs at the injection site during, immediately after, or a combination thereof.

[0693] This disclosure includes the following non-limiting embodiments. [Examples]

[0694] Example 1: Donor commonality analysis: Data-driven donor Z survey A series of hypersensitivity-related adverse event (HSR-AE) reports became known for specific SCIG product lots. Because all hypersensitivity signals received for these IgG lots exhibited similar signals, a data-driven investigation was initiated specifically to determine whether a plasma donor existed that introduced biochemical signals to the end product that could trigger hypersensitivity-related reactions (HSR-AEs) in patients. Hereafter, this donor will be referred to as "Donor Z," and the data-driven investigation as the "Donor Z Investigation."

[0695] methodology approach We assumed that Donor Z was the exclusive cause of the observed HSR-AEs, and that most or all of Donor Z's donations led to the reporting of HSR-AEs in each IgG lot. Under these strong assumptions, it was clear that Donor Z was involved in all retrieved lots. Therefore, only one donor ("common donor") involved in all affected lots was identified.

[0696] In the initial stages of the first four recovery lots, and even in the fifth, sixth, or more recovery lots, hundreds of donors were still common donors, and additional insights were needed to identify Donor Z. To resolve this issue, the following considerations and implications were assumed.

[0697] Based strictly on the assumption of a single donor Z, all other common donors were understood as coincidentally common donors in the sense that their donations coincidentally produced the same final product (FP) as donor Z. 1. The likelihood that any given donor has made donations in all recovered FPs increases strongly with the number of donations made by that donor, all other conditions being equal. This means that most common donors are regular and frequent plasma donors characterized by numerous donations. However, this low relative frequency is hardly consistent with regular and frequent donors, as only a small number of manufactured IgG lots were associated with HSR-AE reporting and voluntary recovery. Although there were only 2.1 donors Z, there were coincidentally several hundred common donors, so a decisive difference had to exist between the two groups, but all the "random" common donors appeared roughly similar. Looking at one or more preferred metrics, one-to-all clustering was expected for the common donors.

[0698] We constructed these metrics and carefully evaluated the evidence and confidence levels of each analysis.

[0699] Scoring of grants A donation was rated "positive" if it was involved in a product formulation (FP) that was voluntarily recalled due to an HSR-AE. A donation was scored "negative" if it was (a) involved in an FP that was not recalled due to an HSR-AE and was completely used up, or (b) was currently on the market, not recalled due to an HSR-AE, and had been on the market long enough that it could be expected that an HSR-AE had already been reported if this lot was the trigger. Specific details regarding the fulfillment of this last condition are provided in the analysis below. All donations that were not scored as "positive" or "negative" were scored as "undetermined." This category includes all donations for which HSR-AE-based scoring is not possible, either because these donations have not yet been processed to FP, manufactured to FP but not yet distributed to patients, or have not been long enough for a return of an HSR-AE report to be expected under normal circumstances.

[0700] metric Based on the scored donations, the following two metrics were calculated separately for each donor.

[0701] The number of negative score assignments per donor is a suitable metric for assessing the number of contraindications regarding a given donor being donor Z. Therefore, it allows for random selection to distinguish between the actual donor Z and all other donors inadvertently included in the retrieved lot. All other things being equal, a lower number of negative score assignments increases the likelihood that a given donor is donor Z.

[0702] The positive test rate (PDR) is,

number

[0703] data Three sets of data were needed to construct a donation trace for all donors associated with the recovered IVIG and SCIG lots. The donation dataset (or dataset 1) provides information about each donation from collection to their pool, as well as additional metadata such as donor and center IDs. The lot trace data (or dataset 2) provides information about the processing of the pool up to the FP stage, in terms of the unique IDs of all intermediates and FPs, as well as date stamps associated with each processing step. In addition, the so-called ex-warehouse date provides information about the date the FP was shipped from the warehouse to its respective market and related entity. The distribution dataset (or dataset 3) provides information about the distribution date, which specifies when the lot was first placed on the market.

[0704] analysis Scoring details The Donor Z study was based on four IgG lots voluntarily recalled due to HSR-AE reporting, requiring all donations to be scored into three categories: positive, negative, and undecided. Specifically, in the FP, the difference between negative and undecided scoring was whether, under normal circumstances, an HSR-AE report for this lot, if present, should have already been received. If yes, the lot was scored as negative; if no, as undecided. To determine this period, the maximum period between the distribution date and the recall date had to be calculated based on the first four recalled lots. This resulted in a result of 88 days, which was considered a conservative estimate of the period typically taken to receive an HSR-AE report for any given IgG lot, given that this period was the maximum across the four lots investigated and also included the period from the receipt of the initial HSR-AE report to the formal recall decision. While most, though not all, lots had a distribution date, the period between the warehouse handover date and recall was calculated as 365 days. This latter estimation was used for scoring the grant only when the distribution date for each lot was unavailable.

[0705] Common donor analysis A common donor analysis was performed on the first four recovered lots, identifying a total of 674 common donors, but no usable insights were obtained. When HSR-AE reports were confirmed for the fifth lot, the common donor analysis was re-evaluated for the five lots with confirmed HSR-AE reports. Across these five lots, a total of 311 common donors remained, and further investigation was deemed necessary to obtain usable insights.

[0706] Table 1 shows the 25 most likely donor Z candidates among the 311 common donors. These 25 most likely donor Z candidates were identified as common donors that met one or more of the following two conditions. ● One of the 15 donors with the fewest negative test results. ● One of the 15 donors with the highest positive donation rate

[0707] Since five donors met both conditions, this resulted in a total of 25 donors for whom donation was proposed to be withheld.

[0708] A more rigorous examination of the 25 donors shown in Table 1 revealed that, as reflected in the positive donation rate and the number of negative donors, there was more evidence that Donor D1 was Donor Z than any of the other 24 Donor Z candidates. This combination made Donor D1 a more of interest as a Donor Z candidate than the other 24 donors and triggered a more rigorous investigation of the timing pattern of donation. [Table 1]

[0709] Verifiable predictions provide confirmation. Based on the presented analysis, the 311 common donors were classified into 286 common donors of lower interest and 25 donors of higher interest over the first five recovery lots. This latter group was further subdivided into 24 moderately interested donors and 1 highly interested donor (donor D1). This classification was purely qualitative and lacked the ability to provide truly probabilistic insights, thus leaving uncertainty as to whether donor D1 was truly donor Z.

[0710] The objective shifted towards confirming candidate donor D1 as donor Z. Ideally, confirmation would come from an independently verifiable prediction, where (a) it can only be stated that one donor meets the relevant criteria after the HSR-AE report has been received, and (b) a true prediction that allows for independent verification and is very unlikely to be correct without having a truly identified donor Z, when there is a clear difference in reliability associated with this.

[0711] As a result, under the pragmatic hypothesis that donor D1 is donor Z, we predicted which lot would be expected to receive the next high level of HSR-AE reporting, and for this purpose, we analyzed the distribution dates of all IgG lots associated with donor D1. By applying the pragmatic hypothesis presented herein, certain lots were predicted to be the next lots expected to receive HSR-AE reporting because they were recently on the market and associated with donor D1.

[0712] Immediately after these predictions were issued, HSR-AE reports were received for all predicted lots. This provided a very high level of confidence in the hypothesis that donor D1 was donor Z, resulting in corresponding prophylactic voluntary recalls and distribution hold for all IgG lots involving plasma involvement from donor D1, respectively.

[0713] Example 2: Functional Test To experimentally support the hypothesis of single plasma donor ("Donor Z") involvement in specific lots of IVIG and SCIG products affected by hypersensitivity adverse events, functional cell-based assays were used to test the relevant product lots, as well as plasma samples from different donors.

[0714] material Instruments, chemicals, and sample preparation All equipment and materials used in this test are reported in Table 2. [Table 2]

[0715] Cryogenic depletion of plasma samples The citrated plasma was thawed at 0°C, and 100 mL was divided into two 50 mL Falcon tubes. The tubes were spun down at 14970 g for 30 minutes at 0°C. The resulting white pellet was collected and weighed between 0.9 and 2.5 g. The supernatant was separated by pipette and stored at -70°C for further analysis.

[0716] Concentrated IgG plasma fraction Donor plasma samples were selectively depleted of IgG using Capture Select® FcXP matrix purchased from Thermo, according to Kober et al. PLo One (2022) 17(1):e0262162. However, the FcXP resin volume was adapted to 1 mL to accommodate the purification of IgG from a 5 mL plasma volume. The resin was washed with an equal volume of PBS, pH 7.4, separated twice by centrifugation, and incubated with the plasma sample on an overhead shaker at 4°C for 60 minutes. After centrifugation at 1000 rpm for 1 minute, the supernatant was collected, and the resin was washed twice by adding 1 mL of PBS, pH 7.4, centrifugation at 1000 rpm for 1 minute, and discarding the supernatant.

[0717] To eluate the bound IgG fraction, the resin was mixed with 1 mL of acetate buffer, pH 4.0 (20 mM acetic acid, 50 mM NaCl), allowed to stand for several minutes, and then centrifuged at 1000 rpm for 1 minute. The supernatant was collected. Elution was repeated 7 to 12 times. The integrity of the elution was evaluated by measuring the IgG concentration in the eluted fraction using a nanodrop spectrophotometer. The capture and elution process was repeated four times with sample plasma using freshly regenerated resin each time. Subsequently, the mixed eluent was concentrated to the initial donor plasma IgG level using a Centriprep centrifuge filter with a 30 kDa cutoff.

[0718] IVIG / SCIG immunoglobulin products The IVIG product tested was a 10% (w / v) plasma-derived polyvalent IgG preparation. The SCIG product tested was a 20% (w / v) plasma-derived polyvalent IgG preparation.

[0719] Decreased IgG plasma fraction Donor plasma samples were selectively reduced for IgE using Capture Select® IgE affinity matrix purchased from Thermo. Resin (0.5–2.5 mL) was washed with an equal volume of PBS, pH 7.4, separated twice by centrifugation, and incubated with 5 mL of plasma sample on an overhead shaker at 4°C for 45 minutes. After centrifugation at 1000 rpm for 1 minute, the supernatant was collected. The reduction procedure was repeated with 0.5 mL of fresh resin and the supernatant.

[0720] IVIG buffer (250mM proline) 450 mL of sterile water for injection was placed in a glass bottle, and 25 mL of 5 M proline solution was added. After mixing, the pH was adjusted to 4.8 by adding 0.2 M HCl (approximately 100 μL), and the volume was increased to 500 mL with sterile water for injection. The solution was then aseptically filtered, filled into a 50 mL glass vial, and sealed with the same stopper under sterile conditions.

[0721] SCIG buffer (250 mM proline, 20 mg / L PS80) 450 mL of sterile water for injection was placed in a glass bottle, and 25 mL of 5 M proline sol...

Claims

1. A method for detecting anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) in a plasma sample or fraction thereof, wherein the method is (i) Contacting the sample with the ligand that binds to the anti-FcεRI Ig in the sample, thereby forming a complex, (ii) A method comprising detecting the complex, wherein the presence of the complex indicates the presence of anti-FcεRI Ig in the plasma sample or a fraction thereof.

2. The method according to claim 1, wherein the anti-FcεRI Ig binds to an epitope of FcεRI and activates an FcεRI-mediated signaling pathway.

3. The method according to claim 1 or 2, wherein the anti-FcεRI Ig binds to the α chain of FcεRI and activates the FcεRI-mediated signaling pathway.

4. The method according to any one of claims 1 to 3, wherein activation of the FcεRI-mediated signaling pathway induces activation and / or degranulation of basophils and / or mast cells.

5. The method according to any one of claims 1 to 4, wherein the anti-FcεRI Ig is anti-FcεRI IgG and / or anti-FcεRI IgE.

6. The method according to any one of claims 1 to 5, wherein the ligand comprises FcεRI or a fragment or epitope thereof.

7. The method according to any one of claims 1 to 6, wherein the detected protein includes an antibody variable region that binds to the anti-FcεRI Ig.

8. The method according to any one of claims 1 to 7, further comprising determining the level of anti-FcεRI Ig in the plasma sample or fraction thereof.

9. The method according to claim 8, wherein the anti-FcεRI IgG is present in a detectable amount in the plasma sample or fraction thereof, and the detectable amount of anti-FcεRI IgG is at least 40 μg / mL.

10. The method according to claim 8 or 9, wherein the anti-FcεRI IgE is present in a detectable amount in the plasma sample or fraction thereof, and the detectable amount of anti-FcεRI IgE is at least 0.1 kU / L.

11. The method according to any one of claims 1 to 10, further comprising determining the percentage of basophil and / or mast cell activation induced by the plasma sample or fraction thereof.

12. The method according to claim 11, wherein the percentage of basophil and / or mast cell activation after exposure to the plasma sample or fraction thereof is determined by determining the percentage of basophils and / or mast cells expressing one or more markers selected from the group consisting of CD63, CD203c, CD107a, and combinations thereof.

13. The method according to claim 12, wherein the proportion of basophils expressing CD63 after exposure to the plasma sample or fraction thereof exceeds 5% of the total population of basophils exposed to the plasma sample or fraction thereof.

14. The method according to claim 12 or 13, wherein the proportion of basophils expressing CD203c after exposure to the plasma sample or fraction exceeds 5% of the total population of basophils exposed to the plasma sample or fraction.

15. The method according to any one of claims 12 to 14, wherein the proportion of mast cells expressing CD107a after exposure to the plasma sample or fraction thereof exceeds 5% of the total population of mast cells exposed to the plasma sample or fraction thereof.

16. The method according to any one of claims 1 to 15, wherein the plasma sample or fraction thereof is selected from the group consisting of human blood plasma sample, IgG intermediate product, intravenous immunoglobulin G (IVIG), subcutaneous immunoglobulin G (SCIG), cryorich plasma, decryoplasma, supernatant I (SN I), Cohn fraction II (Fr II), Cohn fraction II + III (Fr II + III), Cohn fraction I + II + III (Fr I + II + III), Kistler / Nitzchemann precipitate A (KNA), Kistler / Nitzchemann precipitate B (KN B), Kistler / Nitzchemann precipitate of supernatant B (KN B + 1), and combinations thereof.

17. The method according to any one of claims 1 to 16, wherein the plasma sample or fraction thereof is a human blood plasma sample or fraction thereof from one or more subjects.

18. The method according to any one of paragraphs 1 to 17, further comprising administering the plasma sample or a fraction thereof to a subject if the anti-FcεRI Ig is not present in a detectable amount.

19. The method according to any one of paragraphs 1 to 18, further comprising including the plasma sample or a fraction thereof in a pooled blood plasma sample if the anti-FcεRI Ig is not present in a detectable amount.

20. The method according to any one of paragraphs 1 to 19, further comprising excluding the plasma sample or a fraction thereof from a pooled blood plasma sample if the anti-FcεRI Ig is present in a detectable amount.

21. A method for preparing an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) reduction preparation from a plasma sample or fraction thereof, wherein the method is: (i) Loading the plasma sample or fraction into the affinity chromatography resin, which contains a ligand that binds to the anti-FcεRI Ig in the plasma sample or fraction immobilized in the matrix of the affinity chromatography resin, (ii) A method comprising collecting an anti-FcεRI Ig reduction preparation.

22. The method according to claim 21, further comprising detecting the level of anti-FcεRI Ig in the plasma sample or fraction thereof by performing the method according to any one of claims 1 to 24.

23. The method according to claim 21 or 22, wherein the anti-FcεRI Ig-reducing preparation induces reduced activation of the FcεRI-mediated signaling pathway compared to the preparation in which anti-FcεRI Ig is not reduced.

24. The method according to any one of claims 21 to 23, wherein the anti-FcεRI Ig-reducing preparation induces reduced basophil and / or mast cell activation compared to the preparation in which anti-FcεRI Ig is not reduced.

25. The method according to claim 24, wherein the percentage of basophils expressing CD63 after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation, and / or the percentage of basophils expressing CD203c after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI Ig-reducing preparation, and / or the percentage of mast cells expressing CD107a after exposure to the anti-FcεRI Ig-reducing preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI Ig-reducing preparation.

26. The method according to any one of claims 21 to 25, wherein the anti-FcεRI Ig reduction preparation comprises less than 40 μg / mL of anti-FcεRI IgG and / or less than 0.1 kU / L of anti-FcεRI IgE.

27. The affinity chromatography is a continuous affinity chromatography, (i) The first affinity chromatography resin, comprising a ligand immobilized on the matrix of the first affinity chromatography resin, which binds to the anti-FcεRI Ig in the plasma sample or fraction thereof, (ii) The method according to any one of claims 21 to 26, comprising continuous affinity chromatography, the second affinity chromatography resin comprising a matrix of the second affinity chromatography resin containing a blood group A antigen and / or a blood group B antigen.

28. A pharmaceutical composition comprising an anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) reduction preparation prepared by any one of claims 21 to 27.

29. Anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig) depleting IgG preparations containing polyclonal IgG.

30. The anti-FcεRI Ig-reducing IgG preparation according to claim 29, comprising less than 40 μg / mL of anti-FcεRI IgG and / or less than 0.1 kU / L of anti-FcεRI IgE.

31. The anti-FcεRI IgG preparation according to claim 29 or 30, wherein the proportion of basophils expressing CD63 after exposure to the anti-FcεRI IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI IgG preparation, and / or the proportion of basophils expressing CD203c after exposure to the anti-FcεRI IgG preparation is less than 5% of the total population of basophils exposed to the anti-FcεRI IgG preparation, and / or the proportion of mast cells expressing CD107a after exposure to the anti-FcεRI IgG preparation is less than 5% of the total population of mast cells exposed to the anti-FcεRI IgG preparation.

32. A pooled IgG preparation comprising a detectable amount of anti-Fc epsilon receptor I (FcεRI) immunoglobulin (Ig).

33. The pooled IgG preparation according to claim 32, wherein the percentage of basophils expressing CD63 after exposure to the pooled IgG preparation is less than 5% of the total population of basophils exposed to the pooled IgG preparation, and / or the percentage of basophils expressing CD203c after exposure to the pooled IgG preparation is less than 5% of the total population of basophils exposed to the pooled IgG preparation, and / or the percentage of mast cells expressing CD107a after exposure to the pooled IgG preparation is less than 5% of the total population of mast cells exposed to the pooled IgG preparation.

34. A method for treating, preventing, and / or delaying the progression of a condition in a subject associated with immunodeficiency, inflammatory disease, autoimmune disease, or acute infection, wherein the method comprises administering the pharmaceutical composition described in claim 28 or an anti-FcεRI IgG-reducing preparation described in any one of claims 29 to 31.

35. A pharmaceutical composition according to claim 28, or an anti-FcεRI IgG-reducing preparation according to any one of claims 29 to 31, for use in treating, preventing, and / or delaying the progression of a condition associated with immunodeficiency, inflammatory disease, autoimmune disease, or acute infection in a subject.

36. Use of the pharmaceutical composition according to claim 28, or the anti-FcεRI IgG-reducing preparation according to any one of claims 29 to 31, for treating, preventing, and / or delaying the progression of a condition associated with immunodeficiency, inflammatory disease, autoimmune disease, or acute infection in a subject.

37. Use of the pharmaceutical composition according to claim 28, or the anti-FcεRI IgG reduction preparation according to any one of claims 29 to 31, in the manufacture of a pharmaceutical product for treating, preventing, and / or delaying the progression of a condition associated with immunodeficiency, inflammatory disease, autoimmune disease, or acute infection in a subject.

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