Antifungal antibody that binds to beta-glucan

Abstract

The present invention relates to an antibody or an antigen-binding fragment thereof having binding specificity for β-glucan. The production of such monoclonal antibodies and nucleic acids encoding such antibodies are also described.

Description

Technical Field

[0001] The present invention relates to an antibody or an antigen-binding fragment thereof having binding specificity for β-glucan, and to the production of such monoclonal antibodies.

Background Art

[0002] Antibodies are glycoproteins belonging to the immunoglobulin superfamily of proteins. There are five major classes of immunoglobulins, including IgG, IgM, IgA, IgE, and IgD. Antibodies are generally formed from a basic structural unit containing two heavy chains and two light chains. For example, an IgG immunoglobulin molecule contains four polypeptide chains, two of which are identical heavy chains (H) and two of which are identical light chains (K). Each heavy chain contains an N-terminal variable domain (VH) and three constant domains (CH1, CH2, and CH3), and has a hinge region that links CH1 and CH2. Each light chain contains an N-terminal variable domain (VL) and a constant domain (CL), which together associate with the VH and CH1 domains of the heavy chain.

[0003] Antibodies can recognize antigens via the fragment portion of the antigen-binding (Fab) variable region (including the VL, CL, VH, and CH1 domains). Antibodies can activate the immune system through the fragment portion of the crystallizable region (Fc region), which is the tail region of the antibody containing the CH2 and CH3 domains.

[0004] The heavy chain variable region (VH) refers to a fragment of the heavy chain of an antibody that contains three complementarity determining regions (CDRs) inserted between framework regions that form a scaffold to support the CDRs. Similarly, the light chain variable region (VL) contains three CDRs and the framework regions surrounding them.

[0005] CDR is a highly variable amino acid sequence that forms loops within the variable regions of the heavy and light chains with respect to an antibody or its antigen-binding fragment. CDRs can thus interact with antigens. The variable regions of the heavy and light chains each contain three CDRs, herein referred to as CDRH1, CDRH2, and CDRH3 (heavy chain CDRs), and CDRL1, CDRL2, and CDRL3 (light chain CDRs).

[0006] IgG is the major antibody isotype found in blood and extracellular fluid. IgG plays many roles in the immune system, including promoting the aggregation of pathogens, activating the complement system, neutralizing toxins, and promoting antibody-dependent cell-mediated cytotoxicity (ADCC). IgG exists as various subtypes including IgG1, IgG2, IgG3, and IgG4. For the purposes of this embodiment, IgG1 is an exemplary antibody. IgG1 antibodies can be derived from non-human sources, such as any source outside of the antibody repertoire of the human germ line, etc. Antibodies obtained from non-human mammals, such as mice, etc., although not identical, contain heavy chain variable region amino acid sequences that can be generally expressed in humans.

[0007] Thus, even if the sequence is of non-human origin, a sequence that can be expressed as if it were derived from the human antibody repertoire can be regarded as a human sequence. When a non-human antibody, for example, a larger portion such as IgG1, is modified to match what is expressed within the human antibody repertoire, the modified antibody is called "humanized", and the process is called "humanization". From another perspective, humanization refers to retaining a specific portion of a mouse sequence that may include CDRs, and incorporating such sequences into portions of human antibodies, such as the constant regions of the heavy and light chains. This is also often called CDR grafting, where heterologous CDRs are incorporated into the human antibody framework to reduce immunogenicity. Generally, such humanized antibodies show an increase in efficacy and a decrease in overall immunogenicity, inflammation, and other undesirable effects (associated with mouse or other non-human antibodies) that frequently occur in humans.

[0008] β-Glucan is a heterogeneous group of natural polysaccharides found in the cell walls of various organisms, including fungi and bacteria, some of which are pathogenic. Studies suggest that β-glucan may play an immunomodulatory role, and thus antibodies with favorable binding affinity for β-glucan may provide beneficial results through the inhibition of these molecules.

[0009] To date, monoclonal antibodies have been modified to provide binding to β-glucan, but the levels of success have varied. Nevertheless, there remains a significant need for monoclonal antibodies, particularly humanized antibodies, that show improvement in binding to β-glucan in the context of clinical, diagnostic, and therapeutic applications. The embodiments described herein are related to these and other improvements. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0010] The present inventors have developed an antibody or an antigen-binding fragment thereof that targets β-glucan and can be used as an antifungal agent or in the production of an antifungal agent that may have the ability to treat fungal infections.

[0011] The present inventors have surprisingly demonstrated that such antibodies with improved binding to β-glucan can be produced. Surprisingly, it has further been demonstrated that when the antibodies are used as antifungal agents and / or in the treatment of fungal infections, they result in improvements in clinical, therapeutic, and diagnostic outcomes.

Means for Solving the Problems

[0012] In one aspect, the present invention provides (a) CDRL1 of SEQ ID NO: 1, (b) CDRL2 selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 3, (c) CDRL3 of SEQ ID NO: 4 and a light chain variable region comprising the same, and (d) CDRH1 selected from the group consisting of SEQ ID NO: 5 to SEQ ID NO: 7, (e) CDRH2 selected from the group consisting of SEQ ID NO: 8 to SEQ ID NO: 12 and SEQ ID NO: 28, (f) CDRH3 of SEQ ID NO: 13 and a heavy chain variable region comprising the same. The present invention provides an antibody or an antigen-binding fragment thereof comprising the same.

[0013] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO: 14 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 16.

[0014] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 18.

[0015] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 15 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 17.

[0016] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 17.

[0017] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 19.

[0018] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 20.

[0019] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and / or a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 29.

[0020] In one aspect, the present invention provides a humanized antibody or antigen-binding fragment.

[0021] In one aspect, the present invention provides an antibody or an antigen-binding fragment thereof having the ability to bind to β-glucan.

[0022] In a further aspect, the antibody or antigen-binding fragment thereof has the ability to bind to 1,3-β-glucan or 1,6-β-glucan.

[0023] In a further aspect, the antibody or antigen-binding fragment thereof has the ability to bind to laminarin.

[0024] In one aspect, the antibody or antigen-binding fragment thereof can be used as an antifungal agent.

[0025] In a further aspect, the antibody or antigen-binding fragment thereof can be used in the treatment of fungal infections.

[0026] In a further aspect, the antibody or antigen-binding fragment thereof can be used in the manufacture of a medicament for treating fungal infections.

[0027] In a further aspect, the antibody or antigen-binding fragment thereof can be used in the treatment of Aspergillus infections.

[0028] In a further aspect, the antibody or antigen-binding fragment thereof can be used in the treatment of Aspergillus fumigatus infections.

[0029] In a further aspect, the present invention provides a method for treating a fungal infection, comprising the step of administering the antibody or antigen-binding fragment thereof of the present invention.

[0030] In a further aspect, the present invention provides a method for treating an Aspergillus infection, comprising the step of administering the antibody or antigen-binding fragment thereof of the present invention.

[0031] In a further aspect, the present invention provides a method for treating an Aspergillus fumigatus infection, comprising the step of administering the antibody or antigen-binding fragment thereof of the present invention.

[0032] In one aspect, the present invention provides a nucleic acid encoding an antibody or an antigen-binding fragment thereof disclosed herein.

[0033] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 23.

[0034] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 24.

[0035] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 22 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 24.

[0036] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 25.

[0037] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 26.

[0038] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 27.

[0039] In a further aspect, the present invention provides a nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and / or a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 30.

[0040] In one aspect, the present invention provides a nucleic acid encoding an antibody or antigen-binding fragment, wherein the antibody or antigen-binding fragment is humanized.

[0041] In one aspect, the present invention provides a nucleic acid encoding an antibody or antigen-binding fragment, wherein the antibody or antigen-binding fragment has the ability to bind to β-glucan.

[0042] In a further aspect, the nucleic acid encodes an antibody or antigen-binding fragment thereof having the ability to bind to 1,3-β-glucan or 1,6-β-glucan.

[0043] In a further aspect, the nucleic acid encodes an antibody or antigen-binding fragment thereof having the ability to bind to laminarin. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] The drawings and embodiments described herein illustrate the multiple alternative structures, aspects, and characteristics of the multiple embodiments and alternative forms disclosed herein, and are not to be understood as limiting the scope of any of these embodiments and alternative forms.

Figure 1

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Figure 4

Mode for Carrying Out the Invention

[0045] Various preferred characteristics and embodiments of the present invention are described herein as non-limiting examples.

[0046] As used herein and in the appended claims, it should be noted that the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0047] The terms "comprising", "comprises", and "comprised of", as used herein, are synonymous with "including", "includes", "containing", or "contains", are inclusive or non-limiting, and do not exclude additional unrecited members, elements, or steps. The terms "comprising", "comprises", and "comprised of" also include the term "consisting of".

[0048] A numerical range includes the numbers defining the range. Unless otherwise indicated, any nucleic acid sequence is written in the 5'→3' direction from left to right, and an amino acid sequence is written in the amino group→carboxy group direction from left to right.

[0049] The published documents discussed herein merely present that they were disclosed prior to the filing date of the present application. None of the content herein is to be construed as an admission that such published documents constitute prior art with respect to the claims appended hereto. All published documents described herein are incorporated herein by reference.

[0050] The present disclosure is not limited to the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used when practicing or testing embodiments of the present disclosure. Those skilled in the art will understand that all features of the invention disclosed herein can be combined without departing from the scope of the disclosed invention.

[0051] Sequence of an antibody or its antigen-binding fragment In one aspect, the present invention provides (a) CDRL1 of SEQ ID NO: 1, (b) CDRL2 selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 3, (c) CDRL3 of SEQ ID NO: 4 comprising a light chain variable region, and (d) CDRH1 selected from the group consisting of SEQ ID NO: 5-7, (e) CDRH2 selected from the group consisting of SEQ ID NO: 8-12 and SEQ ID NO: 28, (f) CDRH3 of SEQ ID NO: 13 comprising a heavy chain variable region and provides an antibody or its antigen-binding fragment comprising the same.

[0052] In one embodiment, the antibody or antigen-binding fragment thereof of the present invention comprises a light chain variable region containing three light chain variable region CDRs selected from CDRL1 of SEQ ID NO: 1, CDRL2 of SEQ ID NO: 2 or SEQ ID NO: 3, and CDRL3 of SEQ ID NO: 4, and CDRH1 selected from SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7, CDRH2 selected from SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 28, and three heavy chain variable region CDRs selected from CDRH3 of SEQ ID NO: 13.

[0053] The present invention includes "variants" of the CDR regions described below. A "variant" of any given sequence is a sequence in which a particular sequence of residues (whether amino acid residues or nucleic acid residues) has been modified such that the polypeptide or polynucleotide in question retains at least one or all of its intrinsic functions. Variant sequences can be obtained by addition, deletion, substitution, modification, exchange, and / or alteration of at least one residue present in a naturally occurring polypeptide or polynucleotide.

[0054] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof, and (b) CDRL2 of SEQ ID NO: 2 or a variant thereof, and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 5 or a variant thereof, and (e) CDRH2 of SEQ ID NO: 8 or a variant thereof, and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region is included.

[0055] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14.

[0056] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 16.

[0057] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, and the heavy chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 16.

[0058] In one embodiment, the antibody or antigen-binding fragment thereof is (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 2 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 16, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 5 or a variant thereof, CDRH2 of SEQ ID NO: 8 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof comprises or consists of.

[0059] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof, and (b) CDRL2 of SEQ ID NO: 2 or a variant thereof, and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 5 or a variant thereof, and (e) CDRH2 of SEQ ID NO: 10 or a variant thereof, and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region comprises.

[0060] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14.

[0061] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 18.

[0062] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, and the heavy chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 18.

[0063] In one embodiment, the antibody or antigen-binding fragment thereof (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 2 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 18, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 5 or a variant thereof, CDRH2 of SEQ ID NO: 10 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof comprising or consisting of.

[0064] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof, and (b) CDRL2 of SEQ ID NO: 3 or a variant thereof, and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 6 or a variant thereof, and (e) CDRH2 of SEQ ID NO: 9 or a variant thereof, and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region comprising.

[0065] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 15.

[0066] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 17.

[0067] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 15, and the heavy chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 17.

[0068] In one embodiment, the antibody or antigen-binding fragment thereof is (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 15, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 3 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 17, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 6 or a variant thereof, CDRH2 of SEQ ID NO: 9 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof comprising or consisting of.

[0069] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof, and (b) CDRL2 of SEQ ID NO: 2 or a variant thereof, and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 6 or a variant thereof, and (e) CDRH2 of SEQ ID NO: 9 or a variant thereof, and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region comprising.

[0070] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14.

[0071] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 17.

[0072] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, and the heavy chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 17.

[0073] In one embodiment, the antibody or antigen-binding fragment thereof (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 2 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 17, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 6 or a variant thereof, CDRH2 of SEQ ID NO: 9 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof comprises or is composed of.

[0074] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof and (b) CDRL2 of SEQ ID NO: 2 or a variant thereof and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 5 or a variant thereof and (e) CDRH2 of SEQ ID NO: 11 or a variant thereof and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region comprises.

[0075] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 14.

[0076] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 19.

[0077] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 14, and the heavy chain variable region has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 19.

[0078] In one embodiment, the antibody or antigen-binding fragment thereof (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 2 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, the light chain variable region, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 19, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 5 or a variant thereof, CDRH2 of SEQ ID NO: 11 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof, the heavy chain variable region comprises or consists of.

[0079] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof and (b) CDRL2 of SEQ ID NO: 2 or a variant thereof and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 7 or a variant thereof and (e) CDRH2 of SEQ ID NO: 12 or a variant thereof and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region is included.

[0080] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14.

[0081] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 20.

[0082] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, and the heavy chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 20.

[0083] In one embodiment, the antibody or antigen-binding fragment thereof (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 2 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 20, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 7 or a variant thereof, CDRH2 of SEQ ID NO: 12 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof comprising or consisting of.

[0084] In one embodiment, the antibody or antigen-binding fragment thereof is (a) CDRL1 of SEQ ID NO: 1 or a variant thereof and (b) CDRL2 of SEQ ID NO: 2 or a variant thereof and (c) CDRL3 of SEQ ID NO: 4 or a variant thereof comprising a light chain variable region, and (d) CDRH1 of SEQ ID NO: 6 or a variant thereof and (e) CDRH2 of SEQ ID NO: 28 or a variant thereof and (f) CDRH3 of SEQ ID NO: 13 or a variant thereof comprising a heavy chain variable region comprising.

[0085] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14.

[0086] In one embodiment, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 29.

[0087] In one embodiment, the antibody or antigen-binding fragment thereof comprises a light chain variable region and a heavy chain variable region, wherein the light chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, and the heavy chain variable region has an amino acid sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 29.

[0088] In one embodiment, the antibody or antigen-binding fragment thereof (i) A light chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 14, wherein the amino acid sequence comprises CDRL1 of SEQ ID NO: 1 or a variant thereof, CDRL2 of SEQ ID NO: 2 or a variant thereof, and CDRL3 of SEQ ID NO: 4 or a variant thereof, and / or (ii) A heavy chain variable region comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 29, wherein the amino acid sequence comprises CDRH1 of SEQ ID NO: 6 or a variant thereof, CDRH2 of SEQ ID NO: 28 or a variant thereof, and CDRH3 of SEQ ID NO: 13 or a variant thereof comprises or consists of.

[0089] Additional properties of the antibody or antigen-binding fragment thereof In one embodiment, the antibody or antigen-binding fragment thereof can be IgG and all its isotypes.

[0090] In one embodiment, the antibody or antigen-binding fragment thereof can be IgG1 and all its allotypes.

[0091] In one embodiment, the antibody or antigen-binding fragment thereof can be IgG2 and all its allotypes.

[0092] In one embodiment, the antibody or antigen-binding fragment thereof can be IgG3 and all its allotypes.

[0093] In one embodiment, the antibody or antigen-binding fragment thereof can be IgG4 and all of its allotypes.

[0094] In one embodiment, the antibody or antigen-binding fragment thereof can be IgA and all of its isotypes.

[0095] In one embodiment, the antibody or antigen-binding fragment thereof can be IgA1 and its allotypes.

[0096] In one embodiment, the antibody or antigen-binding fragment thereof can be IgA2 and all of its allotypes.

[0097] In one embodiment, the antibody or antigen-binding fragment thereof can be IgM and all of its allotypes.

[0098] In one embodiment, the antibody or antigen-binding fragment thereof can be IgE and all of its allotypes.

[0099] In one embodiment, the antibody or antigen-binding fragment thereof can be IgD and all of its isotypes.

[0100] In one embodiment, the antibody or antigen-binding fragment thereof can be IgD1 and all of its allotypes.

[0101] In one embodiment, the antibody or antigen-binding fragment thereof can be IgD2 and all of its allotypes.

[0102] In one embodiment, the antibody or antigen-binding fragment thereof is a chimeric antibody.

[0103] In a preferred embodiment, the antibody or antigen-binding fragment thereof is a humanized antibody.

[0104] In a preferred embodiment, the antibody or antigen-binding fragment thereof is an affinity matured antibody.

[0105] In a preferred embodiment, the antibody or antigen-binding fragment thereof is an affinity matured humanized antibody.

[0106] Accordingly, the terms "humanized", "humanization", or other forms related to the term "humanize ~" when used in the context of the present disclosure can be a full antibody or any part or fragment thereof, modified to resemble human IgG1, but must be understood to mean an antibody structure that retains at least one CDR of non-human origin. Such modifications can be carried out on mouse IgG antibodies, such as IgG1, but also in some embodiments, the monoclonal antibody of interest is humanized or chimeric. Such modifications can be characterized by one or more amino acid substitutions that match the human germline sequence with respect to the corresponding number of amino acids found in the non-human mammalian form of IgG1. Such substitutions can occur in one or more of the CDRs, or within the framework regions between the CDRs, or indeed anywhere in the variable portions of the light and heavy chains.

[0107] Antibody fragment The present disclosure herein will discuss more about IgG, but the scope of the embodiments is not limited to IgG and includes other naturally occurring antibodies, such as IgM, IgA, IgE, and IgD, etc., as well as their respective subtypes and allotypes. The scope of the embodiments further includes antigen-binding fragments of the above antibodies, including but not limited to, antigen-binding fragments (Fab), fragment antibodies (F(ab’)2), single-chain antibodies (scFv), and single-domain antibodies (sdAb).

[0108] Fragment antibody (F(ab’)2) refers to the region on the antibody remaining after digestion of the Fc region (a part of the hinge region is retained as it is).

[0109] A single-chain antibody (scFv) refers to an antibody that has been modified to be composed of a light-chain variable region and a heavy-chain variable region connected by a peptide linker sequence. The peptide linker sequence generally has a length consisting of 10 to 25 amino acids (rich in glycine to enhance flexibility and rich in serine or threonine to enhance solubility). The peptide linker can connect the N-terminus of the heavy-chain variable region to the C-terminus of the light-chain variable region.

[0110] A single-domain antibody (sdAb), often referred to as a nanobody, refers to an antigen-binding fragment of an antibody composed of a single monomeric variable antibody domain. Thus, an sdAb can be a light-chain variable region or a heavy-chain variable region. Examples of single-domain antibodies include, but are not limited to, VHH fragments and VNAR fragments. VHH and VNAR fragments contain the antigen-binding fragment of the heavy chain.

[0111] In one embodiment, the antigen-binding fragment of the antibody disclosed herein can be any fragment of the antibody disclosed herein.

[0112] In another embodiment, the antigen-binding fragment of the antibody disclosed herein can be a genetically modified product consisting of one or more of the fragments of the antibody.

[0113] Although more discussion in this disclosure is about IgG, the scope of the embodiments is not limited to IgG and includes other naturally occurring antibodies such as IgM, IgA, IgE, and IgD, etc., and their respective subtypes.

[0114] In one embodiment, the fragment is an antigen-binding fragment (Fab), fragment antibody (F(ab’)2), single-chain variable fragment (scFV), or single-domain antibody (sdAb), or camelid antibody (VHH).

[0115] The antibodies or antibody binding fragments disclosed herein can be combined to generate modified antigen-binding proteins that exhibit specificity for two or more different antigens or epitopes. Further, the antibodies or antigen-binding fragments thereof disclosed herein can be combined with alternative antibodies to generate modified antigen-binding proteins that exhibit specificity for two or more different antigens or epitopes. Further, the antibodies or antigen-binding fragments thereof disclosed herein can be combined with one or more identical antibodies, antigen-binding fragments thereof to form homodimers or homotrimers.

[0116] Binding characteristics of antibodies In one embodiment, the antibodies or antigen-binding fragments thereof disclosed herein bind to β-glucan.

[0117] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein bind to 1,3-β-glucan or 1,6-β-glucan.

[0118] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein bind to laminarin.

[0119] The antibodies or antigen-binding fragments thereof disclosed herein can be bispecific. The term bispecific is used to refer to a molecule based on an antibody having two different antigen-binding sites, such as an antibody or antigen-binding fragment thereof or a combination of such. In other words, the antibody-based molecule can be a modified antibody that fits two targets or two epitopes on the same target. Bispecific antibodies are provided in many formats. For example, bispecific antibodies can be generated through a combination of any of the antibodies or antigen-binding fragments thereof disclosed herein and another antigen-binding protein, antibody, or antigen-binding fragment thereof.

[0120] The antibodies or antigen-binding fragments thereof disclosed herein can be multispecific. The term "multispecific" is used to refer to molecules based on antibodies having two or more different antigen-binding sites, such as antibodies or antigen-binding fragments thereof or combinations of such. In other words, the antibody-based molecule can be a modified antibody that fits two or more targets or two or more epitopes on the same target. Multispecific antibodies are provided in many formats. For example, a multispecific antibody can be generated through a combination of at least one of the antibodies or antigen-binding fragments thereof disclosed herein and at least one other antigen-binding protein, antibody, or antigen-binding fragment thereof.

[0121] In one embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in bispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind two different antigens or epitopes.

[0122] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in bispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind β-glucan and an additional antigen or epitope.

[0123] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in bispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind 1,3-β-glucan or 1,6-β-glucan and an additional antigen or epitope.

[0124] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in bispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind laminarin and an additional antigen or epitope.

[0125] In one embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in multispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind two or more different antigens or epitopes.

[0126] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in multispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind two or more different antigens or epitopes.

[0127] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in multispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind β-glucan and one or more different antigens or epitopes.

[0128] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in multispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind 1,3-β-glucan or 1,6-β-glucan, and one or more different antigens or epitopes.

[0129] In a further embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be used in multispecific antigen-binding proteins, antibodies, or antigen-binding fragments thereof that bind laminarin, and one or more different antigens or epitopes.

[0130] In another embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be in the form of a homodimer.

[0131] A homodimer can, by definition, be formed through the binding or interaction of two identical proteins. Thus, in one embodiment, the antibodies or antigen-binding fragments thereof disclosed herein can be linked or interact with another identical antibody or antigen-binding fragment thereof to generate a homodimer.

[0132] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homodimer composed of two antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity.

[0133] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homodimer composed of two antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity for β-glucan.

[0134] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homodimer composed of two antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity for 1,3-β-glucan or 1,6-β-glucan.

[0135] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homodimer composed of two antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity for laminarin.

[0136] In another embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homotrimer.

[0137] By definition, homotrimers can be formed through the binding or interaction of two or three or more identical proteins. Thus, in one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can bind or interact with one or two or more identical antibodies or antigen-binding fragments thereof to generate a homotrimer.

[0138] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homotrimer composed of two or three or more antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity.

[0139] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homomultimer composed of two or more antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity for β-glucan.

[0140] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homomultimer composed of two or more antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity for 1,3-β-glucan or 1,6-β-glucan.

[0141] In one embodiment, the antibody or antigen-binding fragment thereof disclosed herein can be in the form of a homomultimer composed of two or more antibody molecules or antigen-binding fragments thereof having the same antigen-binding specificity for laminarin.

[0142] In one embodiment, the homodimer or homomultimer exhibits an increase in avidity. In other words, the strength of the interaction between the homodimer or homomultimer and the antigen increases.

[0143] The antibody or antigen-binding fragment thereof of the present invention can be defined by its EC50 value. The EC50 value is an indicator of the 50% effective concentration and represents the concentration of the drug required to have 1 / 2 of the maximum effect. The binding property of the antibody or antigen-binding fragment thereof disclosed herein can thus be represented by its EC50 value (determined by any suitable assay method, such as ELISA, etc.). The antibody or antigen-binding fragment thereof disclosed herein can suitably have an EC50 value of about 2 μg / mL or less, about 1 μg / mL or less, about 0.5 μg / mL or less, about 0.4 μg / mL or less, about 0.3 μg / mL or less, about 0.2 μg / mL or less, about 0.1 μg / mL or less, or about 0.05 μg / mL or less when determined by laminarin ELISA.

[0144] Antibody binding affinity is the equilibrium binding constant (K D) can also be determined by, but the identification number can be determined by an appropriate assay method. The antibody or its antigen-binding fragment binds to β-glucan, such as 1,3-β-glucan, 1,6-β-glucan, or laminarin, etc., with a binding affinity (K -8 of 1.5×10 -8 M or less, 1×10 -9 M or less, 9×10 -9 M or less, 8×10 -9 M or less, 7×10 -9 M or less, 6×10 -9 M or less, 4×10 -9 M or less, 3×10 -9 M or less, 2×10 -9 M or less, or 1×10 -9 M or less is appropriate. D ) and can bind.

[0145] The antibody or its antigen-binding fragment can be further defined by its thermal stability. Thermal stability can be defined by the unfolding events that occur when the antibody or its antigen-binding fragment is gradually heated. "T onset " is the temperature at which the unfolding event begins, "T m " is the temperature at the midpoint of the unfolding event, and T agg is the temperature at which the unfolding event causes aggregation. "T m " (the midpoint of the unfolding event) usually occurs as two transition points (referred to as T m1 and T m2 ). The T m1 value corresponds to the first transition point thought to be related to the thermal unfolding of the CH2 domain. The T m2 value corresponds to the second transition point suggesting the unfolding of the CH3 / Fab domain.

[0146] In one embodiment, the antibody or its antigen-binding fragment is heat-resistant up to 60°C.

[0147] In one embodiment, the antibody or its antigen-binding fragment is heat-resistant up to 70°C.

[0148] In one embodiment, the antibody or its antigen-binding fragment is heat-resistant up to 80°C.

[0149] Pharmaceutical preparation In one embodiment, the present invention provides a pharmaceutical composition comprising the antibody of the present invention or its antigen-binding fragment. The antibody or its antigen-binding fragment can be combined with a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.

[0150] In one embodiment, the present invention provides a pharmaceutical composition comprising an antibody or its antigen-binding fragment in combination with L-histidine and sucrose.

[0151] In one embodiment, the present invention provides a pharmaceutical composition comprising 20 mM of L-histidine and 240 mM of sucrose and containing the antibody or its antigen-binding fragment at 5 mg / kg.

[0152] In one embodiment, the present invention provides a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof at least 1 mg / g, at least 2 mg / kg, at least 3 mg / kg, at least 4 mg / kg, at least 5 mg / kg, at least 6 mg / kg, at least 7 mg / kg, at least 8 mg / kg, at least 9 mg / kg, at least 10 mg / kg, at least 15 mg / kg, or at least 20 mg / kg, in combination with at least 5 mM, at least 10 mM, at least 15 mM, at least 16 mM, at least 17 mM, at least 18 mM, at least 19 mM, at least 20 mM, at least 21 mM, at least 22 mM, at least 23 mM, at least 24 mM, at least 25 mM, or at least 30 mM of L-histidine, and at least 200 mM, at least 210 mM, at least 220 mM, at least 230 mM, at least 235, at least 236 mM, at least 237 mM, at least 238 mM, at least 239 mM, at least 240 mM, at least 241 mM, at least 242 mM, at least 243 mM, at least 244 mM, at least 245 mM, at least 250 mM, at least 260 mM, at least 270 mM, at least 280 mM, at least 290 mM, or at least 230 mM of sucrose.

[0153] In one embodiment, the present invention provides an antibody or an antigen-binding fragment thereof according to the present invention, and / or a pharmaceutical composition according to the present invention for use as a medicament.

[0154] In one embodiment, the present invention provides the use of an antibody or an antigen-binding fragment thereof according to the present invention, or a pharmaceutical composition according to the present invention, for the manufacture of a medicament.

[0155] In one embodiment, the present invention provides a method comprising administering an antibody or an antigen-binding fragment thereof according to the present invention, or a pharmaceutical composition according to the present invention, to a subject in need thereof.

[0156] In one embodiment, the antibody or antigen-binding fragment thereof of the present invention can be used as an antifungal agent.

[0157] In one embodiment, a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof can be used as an antifungal agent.

[0158] In one embodiment, the antibody or antigen-binding fragment thereof of the present invention can be used in the treatment of fungal infections.

[0159] In one embodiment, a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof can be used in the treatment of fungal infections.

[0160] In one embodiment, the antibody or antigen-binding fragment thereof or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof can be used in the treatment of Aspergillus infections.

[0161] In one embodiment, the antibody or antigen-binding fragment thereof or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof can be used in the treatment of Aspergillus fumigatus infections.

[0162] In one embodiment, the antibody or antigen-binding fragment thereof or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof can be used in the treatment of Candida infections.

[0163] In one embodiment, the antibody or antigen-binding fragment thereof or a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof can be used in the treatment of Candida albicans or Candida auris infections.

[0164] In one embodiment, the present invention provides a method for treating a fungal infection, comprising the step of administering an antibody or antigen-binding fragment thereof.

[0165] In one embodiment, the present invention provides a method for treating a fungal infection, comprising the step of administering a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof.

[0166] In one embodiment, the present invention provides a method for treating an Aspergillus infection, comprising the step of administering an antibody or an antigen-binding fragment thereof or a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof.

[0167] In one embodiment, the present invention provides a method for treating an Aspergillus fumigatus infection, comprising the step of administering an antibody or an antigen-binding fragment thereof or a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof.

[0168] In one embodiment, the antibody or an antigen-binding fragment thereof of the present invention can be used in the manufacture of a medicament for treating a fungal infection.

[0169] In one embodiment, the pharmaceutical composition comprising the antibody or an antigen-binding fragment thereof of the present invention can be used in the manufacture of a medicament for treating a fungal infection.

[0170] In one embodiment, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof of the present invention can be used in the manufacture of a medicament for treating an Aspergillus infection.

[0171] In one embodiment, the antibody or an antigen-binding fragment thereof or the pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof of the present invention can be used in the manufacture of a medicament for treating an Aspergillus fumigatus infection.

[0172] In one embodiment, the present invention provides a method for treating a Candida infection, comprising the step of administering an antibody or an antigen-binding fragment thereof or a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof.

[0173] In one embodiment, the present invention provides a method for treating Candida albicans infection, comprising the step of administering an antibody or an antigen-binding fragment thereof, or a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof.

[0174] In one embodiment, the present invention provides a method for treating Candida auris infection, comprising the step of administering an antibody or an antigen-binding fragment thereof, or a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof.

[0175] In one embodiment, the antibody or an antigen-binding fragment thereof, or the pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof according to the present invention can be used in the manufacture of a medicament for treating Candida infections.

[0176] In one embodiment, the antibody or an antigen-binding fragment thereof, or the pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof according to the present invention can be used in the manufacture of a medicament for treating Candida albicans infection.

[0177] In one embodiment, the antibody or an antigen-binding fragment thereof, or the pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof according to the present invention can be used in the manufacture of a medicament for treating Candida auris infection.

[0178] Antibody-drug conjugate An antibody-drug conjugate (ADC) is a therapeutic molecule that can specifically target an antigen via an antibody or an antigen-binding fragment thereof, and enables precise delivery of a drug to a predetermined location. For example, an ADC has been used as a chemotherapeutic agent that directs a cytotoxic drug to target antigen-expressing tumor cells, and when internalization of the ADC by the tumor cells occurs, the cytotoxic drug can exert its effect within specific tumor cells. This strategy has been further evolved by conjugating the drug to the antibody through a cleavable linker molecule, enabling cleavage of the ADC once it has internalized into a specific cell, releasing the cytotoxic drug and thus achieving higher activity.

[0179] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein.

[0180] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety.

[0181] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety connected by a cleavable linker molecule.

[0182] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety connected by a non-cleavable linker molecule.

[0183] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety, wherein the coupling moiety is a drug.

[0184] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety, wherein the coupling moiety is a toxin.

[0185] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety, wherein the coupling moiety is a cytokine.

[0186] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety, wherein the coupling moiety is an enzyme.

[0187] In one embodiment, the present invention provides an antibody-drug conjugate comprising an antibody or an antigen-binding fragment thereof disclosed herein and a coupling moiety, wherein the coupling moiety is a drug, toxin, cytokine, enzyme, or a combination thereof.

[0188] Antibody-toxin fusion protein Antibody-toxin fusion proteins, such as recombinant immunotoxins, etc., contain an antibody or an antigen-binding fragment thereof linked to a peptide toxin, or an immunoglobulin. Such antibody-toxin fusion proteins may be encoded by recombinant DNA sequences and then expressed. The recombinant DNA sequences, also known as fusion genes, can be expressed via an expression vector or plasmid containing DNA segments that direct the synthesis of such fusion proteins. Antibody-toxin fusion proteins, or recombinant immunotoxins, thus contain amino acid sequences corresponding to an antibody, an antigen-binding fragment thereof, or an immunoglobulin linked to a peptide toxin.

[0189] In other words, a recombinant immunotoxin is a polypeptide in which a peptide toxin is genetically linked to an antibody component, often by a continuous polypeptide linker.

[0190] Similar to antibody-drug conjugates, antibody-toxin fusion proteins enable the accumulation and activation of targeted toxins to achieve specific effects while minimizing harm to normal cells.

[0191] In one embodiment, the present invention provides a recombinant fusion protein, wherein the fusion protein contains an antibody or an antigen-binding fragment thereof disclosed herein linked to a peptide toxin.

[0192] In one embodiment, the present invention provides a recombinant fusion protein, wherein the fusion protein contains an antibody or an antigen-binding fragment thereof disclosed herein, and at least one of the heavy chain or light chain regions is linked to a peptide toxin.

[0193] In one embodiment, the present invention provides a recombinant fusion protein, wherein the fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, and at least one of SEQ ID NOs: 14-20 and 29 is linked to a peptide toxin.

[0194] In one embodiment, the recombinant fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 16 is linked to a peptide toxin.

[0195] In one embodiment, the recombinant fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 15 and SEQ ID NO: 17 is linked to a peptide toxin.

[0196] In one embodiment, the recombinant fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 18 is linked to a peptide toxin.

[0197] In one embodiment, the recombinant fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 19 is linked to a peptide toxin.

[0198] In one embodiment, the recombinant fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 20 is linked to a peptide toxin.

[0199] In one embodiment, the recombinant fusion protein comprises an antibody or an antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 17 is linked to a peptide toxin.

[0200] In one embodiment, the recombinant fusion protein comprises an antibody or antigen-binding fragment thereof disclosed herein, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 29 is linked to a peptide toxin.

[0201] Nucleotide sequence The antibody or antigen-binding fragment thereof can be encoded by any suitable nucleotide sequence.

[0202] In some embodiments, the nucleotide sequence is codon-optimized, for example, codon-optimized for expression in humans or plants. Cells can vary in their bias towards specific codons, some of which are used more frequently. This codon bias corresponds to the relative abundance of specific tRNAs within a given cell type. Thus, codons are altered within the sequence to match the tRNA with the highest relative abundance within a particular cell type, which can increase the expression level. Similarly, the expression level can be decreased by changing the sequence to match the tRNA with the lowest abundance. Therefore, a higher degree of translational control can be utilized. Codon usage frequencies are known in the art for various organisms.

[0203] In one embodiment, the present invention provides a nucleic acid encoding an antibody or antigen-binding fragment thereof disclosed herein.

[0204] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21.

[0205] In one embodiment, the nucleic acid encodes a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 23.

[0206] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 23.

[0207] In one embodiment, the nucleic acid encodes a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 25.

[0208] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 25.

[0209] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 22.

[0210] In one embodiment, the nucleic acid encodes a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 24.

[0211] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 22, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 24.

[0212] In one embodiment, the nucleic acid encodes a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 26.

[0213] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 26.

[0214] In one embodiment, the nucleic acid encodes a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 27.

[0215] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 27.

[0216] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 24.

[0217] In one embodiment, the nucleic acid encodes a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 30.

[0218] In one embodiment, the nucleic acid encodes a light chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 21, and a heavy chain variable region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 30.

[0219] Expression system The present invention further relates to methods for producing and modifying the antibodies or antigen-binding fragments thereof disclosed herein.

[0220] In some embodiments, a range of production hosts can be used to produce the antibodies or antigen-binding fragments thereof disclosed herein. Such hosts are utilized for the production of antibodies or antigen-binding fragments thereof and can include, but are not limited to, mammalian cells (e.g., CHO cells), plant cells (e.g., Nicotiana benthamiana cells), eukaryotic cells (e.g., yeast cells), and microbial cells.

[0221] In some embodiments within the scope of the present disclosure, as is known in the art, one or more plasmid expression vectors can be conjugated and transfected into living cells for transient expression by a host. When multiple expression vectors are used, they can be transfected into the cell individually or together and expressed by the host. In a preferred embodiment, a modified Nicotiana benthamiana (Nb) plant line is the production host, and one or more expression vectors containing nucleic acids encoding the antibodies or antigen-binding fragments thereof of the present disclosure are transfected therein.

[0222] In one embodiment, the plasmid expression vector is transfected into a host of the genus Agrobacterium. In a preferred embodiment, the plasmid expression vector is transfected into Agrobacterium tumefaciens.

[0223] In a further embodiment, the vector-carrying Agrobacterium is introduced into the plant host by spray inoculation at a slightly abraded site. In a further embodiment, the vector-carrying Agrobacterium is introduced into the plant host by the vacuum infiltration method.

[0224] Further embodiments Monoclonal antibodies containing immunoglobulins have been modified and produced in various types of eukaryotic cells and other living cells. However, mammalian expression systems are often disadvantageous because the production turnaround cycle is slow, and when produced using certain cell lines, there are also constraints because mammalian viruses and other undesirable compounds generated together with the monoclonal antibodies exist within the mammalian system.

[0225] Although not intended to limit the scope of the present embodiment, it should be noted that the plant-based antibody production system can avoid certain pitfalls of mammalian-based monoclonal antibody production. One such system, provided as a non-limiting example, utilizes a modified Nicotiana benthamiana (Nb) plant line that expresses one or more expression vectors containing nucleic acid constructs for forming the desired mAb. In some embodiments within the scope of the present disclosure, as is known in the art, one or more plasmid expression vectors are complexed with viable cells for transient expression by the host and then transfected therein. When multiple expression vectors are used, they can be transfected individually or together into cells such as those found in Nb plant tissue and can be expressed by the host. An exemplary transient expression process uses an Agrobacterium strain (e.g., Agrobacterium tumefaciens), in which case the Agrobacterium grows, infiltrates, infects the plant throughout, and introduces genes into the plant cells. After achieving infiltration and transient expression of the gene of interest in the germinated plant, appropriate nutrient supplementation and care of the infiltrated plant enables growth of the desired plant and ultimately harvesting of an infiltrated plant sample containing the mAb. The desired mAb product is then extracted from the harvested portion of the plant, generally followed by purification and characterization of the extracted monoclonal antibody.

[0226] The manner of introducing vector-bearing Agrobacterium into such a host is not intended as a limiting factor, and spray inoculation or vacuum infiltration into moderately abraded areas are also options. Again, other methods can also be employed with the plant host, and other non-plant hosts are also available with nucleic acid constructs introduced into such hosts through known methods.

[0227] As will be appreciated by those skilled in the art, a range of other production hosts may be suitable for use in mAb production. Such hosts utilized for mAb production according to this embodiment can include, but are not necessarily limited to, mammals (e.g., CHO cells), plants (e.g., Nb plants or plant cells), yeast, and eukaryotic cells, and microbial cells. When going through known steps available in the relevant literature, such hosts have been developed through genetic engineering, mutagenesis, or (in the case of plants) selective breeding for the purpose of enhancing the mAb production amount or providing additional physicochemical characteristics regarding the increase or decrease of molecular, metabolic, chemical, phenotypic characteristics, or other characteristics that affect protein formation by the host.

[0228] This embodiment provides a representative amino acid sequence of a monoclonal antibody derived from a murine source (humanized by modifying the amino acid sequence to increase its similarity to antibody sequence variants found in humans). Thus, while not necessarily limited to the sequences specifically presented herein, amino acid sequences within the scope of this disclosure include humanized monoclonal antibodies that exhibit other activities preferred for neutralizing anti-β-glucan activity or β-glucan effects. In some aspects, the anti-β-glucan activity targets 1,3-β-glucan or 1,6-β-glucan molecules.

[0229] Such amino acid sequences can be provided as the antigen-binding sequences of such antibodies, or in any combination of the above forms or other forms, including fully assembled double-stranded antibodies; single-chain antibodies; antibody fragments comprising an antigen-binding site having the ability to bind to a target molecule and comprising the variable regions of immunoglobulin antibodies, such as IgG, that induce an immune response in an organism, particularly a mammal, including a human; antibody fragments comprising one or more CDRs of the variable regions derived from such antibodies; human antibodies, chimeric antibodies, or humanized antibodies; multimeric antibodies, such as hexameric antibodies, etc. (but not limited thereto); nanobodies; one or more individual immunoglobulin domains; and humanized mAbs in the form of fusion proteins comprising one or more antibodies or antibody portions of the present disclosure.

[0230] Furthermore, embodiments of the present disclosure include any product packaged in any number of forms (e.g., syringes, vials, blister packagings, inhalers). Such products can contain appropriate buffers, additives, and excipients, and can be delivered as unit doses administered through various routes, including subcutaneous, intramuscular, intradermal, injection, oral, nasal, and topical administration, or other forms (any of which can be clinically applicable), but not limited thereto, via syringes, pills, sprays, inhalers, and other means. Such products can be in the form of biological preparations or pharmaceutical compositions having, for example, the antibodies (singular or plural) of the present disclosure coupled to appropriate carriers, such as non-infectious carriers (e.g., viruses, virus-like particles, and nanoparticles, etc.), in a clinically or therapeutically effective amount.

[0231] The pharmaceutical compositions contemplated herein can include monoclonal antibodies as disclosed herein, together with a pharmaceutically acceptable carrier. Also included within the scope of the present disclosure are the mAbs themselves as disclosed herein and methods for manufacturing them.

[0232] In view of the teachings of this specification, various novel mAbs generated based on the present embodiments as described and / or claimed demonstrate several benefits, including, by way of example only, improved affinity for specific antigens or target molecules, increased half-life, increased scalability to achieve higher yields, and cost-effectiveness. Also described herein are several exemplary antibody sequences that target β-glucan (i.e., β-glucan molecules, such as 1,3-β-glucan and 1,6-β-glucan molecules) to achieve these goals.

[0233] This embodiment includes one or two humanized monoclonal antibodies that have the ability to bind to β-glucan molecules and actually bind to them. Five humanized heavy chain amino acid sequences, along with five humanized light chain amino acid sequences, are specifically identified. This embodiment includes several humanized mAbs formed from combinations of heavy and light chains.

[0234] Humanization and Purification of the Targeted mAb Although various humanization techniques have been attempted, many of which are published or otherwise known in the art, there is always a balance in preserving the binding affinity of the animal-derived antibody while achieving appropriate compatibility when the humanized mAb is administered to human subjects. For the amino acid sequences according to the invention disclosed herein, a sequence analysis of the corresponding human acceptor framework was performed for both the heavy and light chain sequences, and the amino acid differences between the human acceptor framework and a known mouse antibody (whose binding activity to β-glucan is determinable) were identified from the results, thereby initiating the humanization process.

[0235] Once the amino acid differences were identified, they were ranked based on the range of biochemical similarities observed between human and mouse subjects. From this step, three variable heavy chain sequences were designed along with three variable light chain sequences. The following table (Table 1) compares the identity of the human sequences to wt (i.e., derived from mouse wild type) as a percentage, which reflects that this percentage is higher than the wt control for all five heavy chain variants.

Table 1

[0236] Factor analysis was performed on the effect of surface-exposed amino acids, and two additional variable heavy chain sequences and two variable light chain sequences were designed by comparing promising variants to similar human antibody structures within Clinical NSG (Next Generation Sequencing Database). For each of these variants, the goal was to increase the number of amino acid substitutions that are identical to the human acceptor framework, as well as the number of conserved amino acids derived from the mouse source as found in the antigen-binding site.

[0237] In addition, data similarly obtained for the variable light chain, showing the same trend in variants and controls, are presented in Table 2.

Table 2

[0238] Accordingly, five heavy chain sequences and five light chain sequences (expressed as pairs in VH and VL, the sequences of which are disclosed herein as SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 29, and SEQ ID NO: 31 to SEQ ID NO: 37) were identified. Without intending to limit these teachings to a single approach, each humanized mAb tested as discussed herein was generated by expressing the heavy chain sequence on a first expression vector and the light chain sequence on a second expression vector. For purposes of non-limiting illustration, a vector containing a potato virus X backbone (PVX) and a vector containing a tomato vein clearing virus backbone (TVCV) are suitable vectors for these purposes.

[0239] Once generated, the monoclonal antibodies of the present disclosure can be purified by methods known in the relevant art. For example, U.S. Patent No. 10,214,747 (Method for Purifying Monoclonal Antibodies, filed Feb. 26, 2019), the contents of which are hereby incorporated by reference in their entirety, describes a monoclonal antibody purification platform that includes procedures and processes for purifying a variety of different antibodies (suitable for purifying the antibodies generated based on the present disclosure).

[0240] Additional amino acid sequences based on this embodiment The exemplary amino acid sequences described herein can be expressed in any of several optional ways and with any arbitrary combination of heavy and light chain amino acid chains. Although exemplary sequences are presented, the scope of the present disclosure is recognized to include portions of the identified sequences, sequences that exhibit general structural characteristics, and other sequences including longer sequences that include the exemplary sequences presented herein.

[0241] The sequences disclosed in this specification include SEQ ID NO: 14, SEQ ID NO: 17, SEQ ID NO: 29, and SEQ ID NOs: 31 to 37 (see Table 3 below), SEQ ID NO: 17, SEQ ID NO: 29. SEQ ID NOs: 31 to 33 are heavy chain amino acid sequences. SEQ ID NO: 14 and SEQ ID NOs: 34 to 37 are light chain amino acid sequences.

[0242] According to this embodiment, the heavy chain of the humanized monoclonal antibody includes the amino acid sequence contained in SEQ ID NO: 17. The amino acid sequence of SEQ ID NO: 17 can form a humanized antibody in combination with a light chain amino acid sequence. In that case, the light chain includes the amino acid sequence contained in any one of SEQ ID NO: 14 and SEQ ID NOs: 34 to 37. As a non-limiting example, the combination of the heavy chain and the light chain can use SEQ ID NO: 17 in combination with SEQ ID NO: 35, or SEQ ID NO: 17 in combination with SEQ ID NO: 14.

[0243] Also, embodiments based on the present disclosure include those that include an amino acid sequence such that the heavy chain of the humanized monoclonal antibody is contained in SEQ ID NO: 31. The amino acid sequence of SEQ ID NO: 31 can form a humanized antibody in combination with a light chain amino acid sequence. In that case, the light chain includes the amino acid sequence contained in any one of SEQ ID NO: 14 and SEQ ID NOs: 34 to 37. As a non-limiting example, the combination of the heavy chain and the light chain can use SEQ ID NO: 31 in combination with SEQ ID NO: 35, or SEQ ID NO: 31 in combination with SEQ ID NO: 14.

[0244] Embodiments based on the present disclosure include those that include an amino acid sequence such that the heavy chain of the humanized monoclonal antibody is contained in SEQ ID NO: 32. The amino acid sequence of SEQ ID NO: 32 can form a humanized antibody in combination with a light chain amino acid sequence. In that case, the light chain includes the amino acid sequence contained in any one of SEQ ID NO: 14 and SEQ ID NOs: 34 to 37. As a non-limiting example, the combination of the heavy chain and the light chain can use SEQ ID NO: 32 in combination with SEQ ID NO: 35, or SEQ ID NO: 32 in combination with SEQ ID NO: 14.

[0245] Furthermore, embodiments of the present disclosure include those that include an amino acid sequence such that the heavy chain of the humanized monoclonal antibody includes the amino acid sequence contained in SEQ ID NO: 29. The amino acid sequence of SEQ ID NO: 29 can form a humanized antibody in combination with a light chain amino acid sequence, in which case the light chain includes an amino acid sequence contained in any of SEQ ID NO: 14 and SEQ ID NO: 34 to SEQ ID NO: 37. As a non-limiting example, a combination of a heavy chain and a light chain can use SEQ ID NO: 29 in combination with SEQ ID NO: 35, or SEQ ID NO: 29 in combination with SEQ ID NO: 14.

[0246] In addition, embodiments of the present disclosure include those that include an amino acid sequence such that the heavy chain of the humanized monoclonal antibody includes the amino acid sequence contained in SEQ ID NO: 33. The amino acid sequence of SEQ ID NO: 33 can form a humanized antibody in combination with a light chain amino acid sequence, in which case the light chain includes an amino acid sequence contained in any of SEQ ID NO: 14 and SEQ ID NO: 34 to SEQ ID NO: 37. As a non-limiting example, a combination of a heavy chain and a light chain can use SEQ ID NO: 33 in combination with SEQ ID NO: 35, or SEQ ID NO: 33 in combination with SEQ ID NO: 14.

[0247] Also, embodiments of the present disclosure also include those that include an amino acid sequence such that the light chain of the humanized monoclonal antibody includes the amino acid sequence contained in SEQ ID NO: 35. The amino acid sequence of SEQ ID NO: 36 can form a humanized antibody in combination with a heavy chain amino acid sequence, in which case the heavy chain includes an amino acid sequence contained in any of SEQ ID NO: 17, SEQ ID NO: 29, and SEQ ID NO: 31 to SEQ ID NO: 33.

[0248] In addition, embodiments of the present disclosure include those that include an amino acid sequence such that the light chain of the humanized monoclonal antibody includes the amino acid sequence contained in SEQ ID NO: 14. The amino acid sequence of SEQ ID NO: 14 can form a humanized antibody in combination with a heavy chain amino acid sequence, in which case the heavy chain includes an amino acid sequence contained in any of SEQ ID NO: 17, SEQ ID NO: 29, and SEQ ID NO: 31 to SEQ ID NO: 33.

[0249] Furthermore, embodiments of the present disclosure include those in which the light chain of the humanized monoclonal antibody includes an amino acid sequence such as those included in SEQ ID NO: 34 and any of SEQ ID NO: 36 to SEQ ID NO: 37. Any of the amino acid sequences of SEQ ID NO: 34 and SEQ ID NO: 36 to SEQ ID NO: 37 can form a humanized antibody in combination with the heavy chain amino acid sequence, in which case the heavy chain includes an amino acid sequence included in any of SEQ ID NO: 17, SEQ ID NO: 29, and SEQ ID NO: 31 to SEQ ID NO: 33.

[0250] Based on the sequences identified above, SEQ ID NO: 17, SEQ ID NO: 29, and SEQ ID NO: 31 to SEQ ID NO: 33 are variants of the heavy chain variable region. Therefore, Table 3 associates each heavy chain variable region variant (VH) with one of the above heavy chain sequence numbers.

[0251] Also, based on the sequences identified above, SEQ ID NO: 14 and SEQ ID NO: 34 to SEQ ID NO: 37 are variants of the light chain variable region. Therefore, Table 3 associates each light chain variant (VK) with one of the above light chain sequence numbers. In view of the above, three clones (3.1, 14.1, and 32.3) are presented herein.

[0252] Clone 3.1 is VH_wt combined with an mAb derived from VL_wild type mice used as a control, and thus none of the above sequences are included in this clone.

[0253] Clone 14.1 is further specified as VH_V1_VL_V5 and includes the combination of SEQ ID NO: 17_ SEQ ID NO: 14.

[0254] Clone 32.3 is further specified as VH_V4_VK_V5 and includes the combination of SEQ ID NO: 29_ SEQ ID NO: 14.

[0255] The embodiments described herein are to be understood as not being limited to the details of the teachings and explanations shown or illustrated in the accompanying drawings when applied. Rather, these embodiments and alternatives are to be understood as capable of being practiced or implemented in various ways as described and claimed herein. Also, the words and phrases used herein are for the purpose of explanation and should not be regarded as limiting. As used herein, "including", "comprising", "e.g.", "containing", or "having", and variations of such words are intended to include the recited items, their equivalents, and additional items.

[0256] Accordingly, the foregoing description of some embodiments and alternatives is intended more as an illustration than as a limitation on the scope of what is disclosed herein. The description herein is not intended to be exhaustive, nor is it intended that the understanding of the embodiments be limited to the exact forms disclosed. Modifications and variations of these embodiments will be understood by those skilled in the art as reasonably possible in light of the above teachings and explanations.

[0257] Embodiments with numbers: 1. (a) CDRL1 of SEQ ID NO: 1, (b) CDRL2 selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 3, (c) CDRL3 of SEQ ID NO: 4 and a light chain variable region comprising, (d) CDRH1 selected from the group consisting of SEQ ID NO: 5 - 7, (e) CDRH2 selected from the group consisting of SEQ ID NO: 8 - 12 and SEQ ID NO: 28, (f) CDRH3 of SEQ ID NO: 13 and a heavy chain variable region comprising an antibody or an antigen - binding fragment thereof. 2. The light chain variable region is (a) CDRL1 of SEQ ID NO: 1, and (b) CDRL2 of SEQ ID NO: 2, and (c) CDRL3 of SEQ ID NO: 4 and The antibody or antigen-binding fragment thereof according to Embodiment 1, comprising 3. The antibody or antigen-binding fragment thereof according to Embodiment 1 or Embodiment 2, wherein the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 14. 4. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 5, and (b) CDRH2 of SEQ ID NO: 8, and (c) CDRH3 of SEQ ID NO: 13 and The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3, comprising 5. The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 4, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 16. 6. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 5, and (b) CDRH2 of SEQ ID NO: 10, and (c) CDRH3 of SEQ ID NO: 13 and The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3, comprising 7. The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3 or 6, wherein the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 18. 8. The light chain variable region is (a) CDRL1 of SEQ ID NO: 1, and (b) CDRL2 of SEQ ID NO: 3, and (c) CDRL3 of SEQ ID NO: 4 and The antibody or antigen-binding fragment thereof according to Embodiment 1, comprising 9. The antibody or antigen-binding fragment thereof according to Embodiment 1 or Embodiment 8, wherein the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 15. 10. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 6, and (b) The CDRH2 of SEQ ID NO: 9, and (c) The CDRH3 of SEQ ID NO: 13, and The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3, 8, or 9. 11. The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3, 8, 9, or 10, wherein the heavy-chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 17. 12. The heavy-chain variable region is (a) The CDRH1 of SEQ ID NO: 5, and (b) The CDRH2 of SEQ ID NO: 11, and (c) The CDRH3 of SEQ ID NO: 13, and The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3. 13. The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3 or 12, wherein the heavy-chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 19. 14. The heavy-chain variable region is (a) The CDRH1 of SEQ ID NO: 7, and (b) The CDRH2 of SEQ ID NO: 12, and (c) The CDRH3 of SEQ ID NO: 13, and The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3. 15. The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3 or 14, wherein the heavy-chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 20. 16. The heavy-chain variable region is (a) The CDRH1 of SEQ ID NO: 6, and (b) The CDRH2 of SEQ ID NO: 28, and (c) The CDRH3 of SEQ ID NO: 13, and The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3. 17. The antibody or antigen-binding fragment thereof according to any one of Embodiments 1 to 3 or 16, wherein the heavy-chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 29. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO: 14. An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 16. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 16. An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 18. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 18. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO: 15. An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 17. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 15 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 17. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 17. An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 19. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 19. 29. An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 20. 30. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 20. 31. An antibody or an antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 29. 32. An antibody or an antigen-binding fragment thereof comprising a light chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 14 and a heavy chain variable region comprising an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 29. 33. The antibody or an antigen-binding fragment thereof according to any one of Embodiments 3, 5, 7, 9, 11, 13, 15, and 17 to 32, having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% sequence identity to the listed sequences. 34. The antibody or an antigen-binding fragment thereof according to any one of Embodiments 1 to 33, which is a single-chain variable fragment (scFV), a single-domain antibody (sdAb), an antigen-binding fragment (Fab), or a fragment antibody (F(ab’)2). 35. The antibody or an antigen-binding fragment thereof according to any one of Embodiments 1 to 34, which is of the IgG, IgA, IgM, IgE, or IgD isotype, or any allotype thereof. 36. The antibody or an antigen-binding fragment thereof according to Embodiment 35, which is of the IgG1 isotype or any allotype thereof. 37. The antibody or an antigen-binding fragment thereof according to Embodiment 35, which is of the IgG2 isotype or any allotype thereof. 38. The antibody or antigen-binding fragment thereof according to embodiment 35, which is of the IgG3 isotype or any allotype thereof. 39. The antibody or antigen-binding fragment thereof according to embodiment 35, which is of the IgG4 isotype or any allotype thereof. 40. The antibody or antigen-binding fragment thereof according to embodiment 35, which is of the IgA1 isotype or any allotype thereof. 41. The antibody or antigen-binding fragment thereof according to embodiment 35, which is of the IgA2 isotype or any allotype thereof. 42. The antibody or antigen-binding fragment thereof according to embodiment 35, which is of the IgD1 isotype or any allotype thereof. 43. The antibody or antigen-binding fragment thereof according to embodiment 35, which is of the IgD2 isotype or any allotype thereof. 44. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 43, which is humanized. 45. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 44, wherein the antibody binds to β-glucan. 46. The antibody or antigen-binding fragment thereof according to embodiment 45, wherein the β-glucan is 1,3-β-glucan or 1,6-β-glucan. 47. The antibody or antigen-binding fragment thereof according to embodiment 45 or embodiment 46, wherein the antibody binds to laminarin. 48. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 47, for use in a bispecific antigen-binding protein, antibody, or antigen-binding fragment thereof that binds two different antigens or epitopes. 49. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 48, for use in a multispecific antigen-binding protein, antibody, or antigen-binding fragment thereof that binds two or more different antigens or epitopes. 50. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 49, which is in the form of a homodimer or homotrimer. 51. The antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 50, which has an EC50 of less than 0.2 μg / mL. An antibody or antigen-binding fragment thereof according to embodiment 51, having an EC50 of less than 0.1 μg / mL. 53. 1.5×10 -8 A value lower than M, of the antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 52. D 54. 1×10 -8 A value lower than M, of the antibody or antigen-binding fragment thereof according to embodiment 53. D 55. 5×10 -9 A value lower than M, of the antibody or antigen-binding fragment thereof according to embodiment 54. D 56. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 55, which is heat-resistant up to 80°C. 57. An antibody or antigen-binding fragment thereof according to embodiment 56, which is heat-resistant up to 70°C. 58. An antibody or antigen-binding fragment thereof according to embodiment 57, which is heat-resistant up to 60°C. 59. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 58, for use as an antifungal agent. 60. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 59, for use in the treatment of a fungal infection. 61. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 60, for use in the manufacture of a medicament for treating a fungal infection. 62. A method for treating a fungal infection, comprising the step of administering an antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 61. 63. An antibody or antigen-binding fragment thereof for use according to embodiments 59 to 61, or a method according to embodiment 62, wherein the fungal infection is an Aspergillus infection. 64. An antibody or antigen-binding fragment thereof, or a method according to embodiment 63, wherein the Aspergillus infection is an Aspergillus fumigatus infection. 65. An antibody or antigen-binding fragment thereof for use according to embodiments 59 to 61, or a method according to embodiment 62, wherein the fungal infection is a Candida infection. ​​​66. The antibody or antigen-binding fragment thereof, or method according to embodiment 65, wherein the Candida infection is a Candida albicans or Candida auris infection. 67. A recombinant fusion protein, wherein the fusion protein comprises an antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 66 and a peptide toxin. 68. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 16 is linked to a peptide toxin. 69. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 15 and SEQ ID NO: 17 is linked to a peptide toxin. 70. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 18 is linked to a peptide toxin. 71. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 19 is linked to a peptide toxin. 72. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 20 is linked to a peptide toxin. 73. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 17 is linked to a peptide toxin. 74. The recombinant fusion protein according to embodiment 67, wherein at least one of SEQ ID NO: 14 and SEQ ID NO: 29 is linked to a peptide toxin. 75. An antibody or antigen-binding fragment thereof according to any one of embodiments 1 to 74, conjugated to a coupling moiety. 76. The antibody or antigen-binding fragment thereof according to embodiment 75, wherein the coupling moiety is a drug. 77. The antibody or antigen-binding fragment thereof according to embodiment 75, wherein the coupling moiety is a toxin. 78. The antibody or antigen-binding fragment thereof according to embodiment 75, wherein the coupling moiety is a cytokine. 79. The antibody or antigen-binding fragment thereof according to embodiment 75, wherein the coupling moiety is an enzyme. 80. An antibody or an antigen-binding fragment thereof according to any one of embodiments 75 to 79, wherein the antibody or the antigen-binding fragment thereof and a coupling moiety are conjugated by a linker molecule. 81. An antibody or an antigen-binding fragment thereof according to embodiment 80, wherein the linker molecule is a cleavable linker molecule. 82. A nucleic acid encoding an antibody or an antigen-binding fragment thereof according to any one of embodiments 1 to 81. 83. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21. 84. A nucleic acid encoding a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 23. 85. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 23. 86. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 22. 87. A nucleic acid encoding a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 24. 88. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 22 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 24. 89. A nucleic acid encoding a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 25. 90. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 25. 91. A nucleic acid encoding a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 26. 92. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 26. 93. A nucleic acid encoding a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 27. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 27. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 24. A nucleic acid encoding a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 30. A nucleic acid encoding a light chain variable region having at least 70% sequence identity to SEQ ID NO: 21 and a heavy chain variable region having at least 70% sequence identity to SEQ ID NO: 30. A nucleic acid according to any one of embodiments 83 to 97, having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% sequence identity to the listed sequences. A compound that is an anti-β-glucan antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region (VH) and a light chain variable region (VL), the VH amino acid sequence is selected from any one of SEQ ID NO: 29 and SEQ ID NOs: 31 to 34, and the VL amino acid sequence is selected from any one of SEQ ID NO: 14 and SEQ ID NOs: 35 to 38. The compound according to embodiment 99, wherein the VH amino acid sequence is either SEQ ID NO: 31 or SEQ ID NO: 29, and the VL amino acid sequence is SEQ ID NO: 14.

[0258]

Table 3

[0259] The present invention will be further described here by way of example, but it is intended to assist those skilled in the art in practicing the present invention and does not include any intention to limit the scope of the present invention in any case.

[0260] The antibodies tested in the following examples, after undergoing two-step chromatography purification, had a purity exceeding 99% when expressed as the final monomer percentage of column HPLC SEC as shown in Table 4.

Table 4

Example

[0261] Laminarin-binding (ELISA) for determining the EC50 values of the initial humanized antifungal antibody and the next-generation affinity-matured antifungal antibody The EC50 value is an indicator of the 50% effective concentration. As detailed below, the EC50 was calculated using laminarin-binding ELISA.

[0262] The ELISA plates were coated overnight at 5 °C with PBS containing 10 μg / mL laminarin (laminarin from Laminaria digitata, Sigma L9634), and then blocked for 2 - 3 hours at room temperature in PBST containing 3% BSA. Serial three-fold diluted antibodies (2G8 variant) starting from 10 μg / mL were applied to the plates and incubated for 1 hour at room temperature. The laminarin-binding antibodies were detected at a 1:20,000 dilution with HRP-conjugated rabbit anti-human IgG FC (Invitrogen, Rockford, IL). Tetramethylbenzidine (TMB) substrate (SeraCare Life Sciences INC, Milford, MA, USA) was used for detection; the absorbance at 450 nm was measured on a plate reader. A graph was plotted (Figure 1), and the EC50 was calculated using GraphPad Prism 8.0 software (Table 5). Two antibodies from KBio (not 2G8) were used as negative controls, and no binding to laminarin was observed.

Table 5

Example

[0263] Determination of the binding affinity (K D ) of the initial humanized antifungal antibody and the next-generation affinity-matured antifungal antibody for laminarin (Biacore-SPR) The equilibrium binding constant (K D ) is an indicator of the binding affinity of an antibody for its target and can be determined by any suitable assay method, such as surface plasmon resonance (SPR), etc.

[0264] The binding affinity (K D) was measured at ambient temperature on a Biacore T200 instrument. For each cycle, according to the manufacturer's instructions, variants of 2G8 were captured on a Pro A sensor chip to a surface density of approximately 1300 - 1500 RU. Serial dilutions of laminarin were made in running buffer (HBS-EP+, Cytiva) and injected at a flow rate of 30 μL / min with a contact time of 120 seconds and a dissociation time of 600 seconds. Between sample injections, the system was washed with Protein A wash buffer. The reference flow cell was used to correct for contributions to the response, such as bulk shifts that occur similarly in the sample and reference flow cells. A blank cycle (running buffer) was performed, and all sample injections were blanks subtracted to correct the sensorgram for drift and other interferences that affect the reference subtracted curve. In each experiment, replicates and blanks of laminarin with non-zero concentrations were injected, and double referencing was performed to verify the reliability of the Pro A chip throughout the experiment. The data was evaluated by kinetic binding analysis for the association rate constant (ka) (Figure 2B; Table 6), the dissociation rate constant (kd) (Figure 2C; Table 7), and the equilibrium dissociation constant (K D )(Figure 2A; Table 8).

Table 6

Table 7

Table 8

Example

[0265] Aspergillus fumigatus induction test in immunosuppressed mice after treatment with initial humanized anti-fungal antibodies or next-generation humanized and affinity matured antibodies, and fungal burden assessment In a mouse model of invasive aspergillosis, animals were first subjected to an immunosuppressive regimen. On day 2 (relative to the inoculation on day 0), all animals were administered cyclophosphamide (250 mg / kg) by intraperitoneal injection and cortisone acetate (500 mg / kg) by subcutaneous administration. On day 3, all animals were administered cyclophosphamide (250 mg / kg) by intraperitoneal injection and cortisone acetate (500 mg / kg) by subcutaneous administration. Body weights were recorded before administration on days 2 and 3, and the animals were administered at a volume of 10 mL per kg.

[0266] At 24 hours and 120 hours after the Aspergillus induction treatment, all test groups were administered 15 mg / kg of mAb (formulated in 20 mM L-histidine + 240 mM sucrose, pH 6.0). The vehicle control group received administration of the formulation buffer (20 mM L-histidine + 240 mM sucrose, pH 6.0). For all groups, the test item (mAb or vehicle) was administered at 2.5 mL per kg.

[0267] Immunosuppressed mice were monitored until 14 days after infection with Aspergillus fumigatus, and the survival rate (%) was recorded daily.

[0268] As expected, the survival rate of mice that received the induction treatment with A. fumigatus and received only the control vehicle was 0% on day 8.

[0269] The survival rate of the mouse groups administered the vehicle control or clone 14.1 antibody was 0% after 8 days. The mouse groups administered the clone 39.1 or 41.2 antibody achieved a survival rate of 10%, and the mouse groups administered the clone 3.1, 32.3, 40.1, 42.1, or 43.1 antibody achieved a survival rate of 20% (Figure 3A).

[0270] The galactomannan level within the BAL was analyzed by ELISA according to the manufacturer's instructions (Platelia Aspergillus Ag #62794, Bio-Rad).

[0271] Histopathological assessment was performed on lung samples after the BAL procedure. Briefly, the animal's chest was opened, the lungs were excised and weighed. The entire lungs were then inflated with 10% neutral buffered formalin for at least 24 hours, after which the tissue was processed for paraffin blocks. Sections of approximately 4 - 5 μm of lung tissue were stained with haematoxylin and eosin (H&E), and Periodic Acid Schiff (PAS). Microscopic evaluation of the lungs included grading of inflammatory cell infiltration and tissue inflammation (by H&E), with special consideration for the presence of fungal spores and hyphae (by PAS).

[0272] Data for the majority of animals in groups 2 - 10 were euthanized prior to 14 days for welfare reasons, and those animals showed detectable levels of fungi in the lungs by either BAL galactomannan levels or PAS positive staining, or both (Figure 3B). Only 2 animals (animal 39 in group 32.3 and animal 47 in group 39.1) did not fall into this category.

[0273] None of the surviving animals in the saline / media groups, 42.1 or 3.1, had detectable levels of fungi in the lungs by either BAL galactomannan levels or PAS positive staining. One of the surviving animals in group 32.3 was positive for PAS staining, while the others were negative. The only surviving animal in group 39.1 had a positive galactomannan BAL, and of the 2 surviving animals in group 40.1, one had a positive galactomannan BAL while the other was negative. The only surviving animal in group 41.2 had a positive galactomannan BAL, and of the 2 surviving animals in group 43.1, one was positive for PAS staining while the other had no detectable levels in either assay.

Example

[0274] Thermal Shift Assay (TSA) for determining the melting temperature of the antibody A Thermal Shift Assay (TSA) was performed to determine the melting temperature of the antibody. The melting temperature (T m ) of the antibody was determined by TSA performed on a qPCR system from Life technologies. Each antibody with a final concentration of 0.5 mg / mL in Protein Thermal Shift™ buffer (Applied Biosystems, catalog number 4462263) was mixed with 1× Protein Thermal Shift™ dye (Applied Biosystems, catalog number 4462263) in a MicroAmp™ Optical 96-well plate to a total volume of 20 μl per well. A blank control was set for the protein alone. The samples were analyzed in triplicate. The plate was heated from 25 °C to 99 °C in 0.05 °C / sec increments. The data were analyzed using Protein Thermal Shift software version 1.4 (Applied Biosystems, catalog number 4466037). T m1 (Figure 4A; Table 9) and T m2 (Figure 4B; Table 10), the melting temperature (T m ) was plotted and displayed.

Table 9

Table 10

Claims

1. (a) CDRL1 of SEQ ID NO: 1, (b) CDRL2 selected from the group consisting of SEQ ID NO: 2 and SEQ ID NO: 3, (c) CDRL3 of SEQ ID NO: 4 and a light chain variable region comprising, and (d) CDRH1 selected from the group consisting of SEQ ID NO: 5 to SEQ ID NO: 7, (e) CDRH2 selected from the group consisting of SEQ ID NO: 8 to SEQ ID NO: 12 and SEQ ID NO: 28, (f) CDRH3 of SEQ ID NO: 13 and a heavy chain variable region comprising An antibody or an antigen-binding fragment thereof comprising.

2. The light chain variable region is (a) CDRL1 of SEQ ID NO: 1, (b) CDRL2 of SEQ ID NO: 2, (c) CDRL3 of SEQ ID NO: 4 and optionally, the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 14, the antibody or antigen-binding fragment thereof according to claim 1.

3. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 5, (b) CDRH2 of SEQ ID NO: 8, (c) CDRH3 of SEQ ID NO: 13 and optionally, the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 16, the antibody or antigen-binding fragment thereof according to claim 1 or 2.

4. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 5, (b) CDRH2 of SEQ ID NO: 10, (c) CDRH3 of SEQ ID NO: 13 and optionally, the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 18, the antibody or antigen-binding fragment thereof according to claim 1 or 2.

5. The light chain variable region is (a) CDRL1 of SEQ ID NO: 1, (b) CDRL2 of SEQ ID NO: 3, (c) CDRL3 of SEQ ID NO: 4 and optionally, the light chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 15, the antibody or antigen-binding fragment thereof according to claim 1.

6. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 6, (b) CDRH2 of SEQ ID NO: 9, (c) CDRH3 of SEQ ID NO: 13 and optionally, the heavy chain variable region comprises an amino acid sequence having at least 70% identity to SEQ ID NO: 17, the antibody or antigen-binding fragment thereof according to any one of claims 1, 2 and 5.

7. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 5, (b) CDRH2 of SEQ ID NO: 11, (c) CDRH3 of SEQ ID NO: 13 The antibody or antigen-binding fragment thereof according to claim 1 or 2, comprising, optionally, a heavy chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO:

19.

8. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 7, (b) CDRH2 of SEQ ID NO: 12, and (c) CDRH3 of SEQ ID NO: 13 The antibody or antigen-binding fragment thereof according to claim 1 or 2, comprising, optionally, a heavy chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO:

20.

9. The heavy chain variable region is (a) CDRH1 of SEQ ID NO: 6, (b) CDRH2 of SEQ ID NO: 28, and (c) CDRH3 of SEQ ID NO: 13 The antibody or antigen-binding fragment thereof according to claim 1 or 2, comprising, optionally, a heavy chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO:

29.

10. The antibody or antigen-binding fragment thereof according to any one of claims 2 to 9, having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% sequence identity to the listed sequences.

11. A single-chain variable fragment (scFv), single-domain antibody (sdAb), antigen-binding fragment (Fab), or fragment antibody (F(ab')) ２ The antibody or antigen-binding fragment thereof according to any one of claims 1 to 10, which is

12. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 11, which is an IgG, IgA, IgM, IgE, or IgD isotype, or any allotype thereof.

13. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 12, which is humanized.

14. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, wherein the antibody binds to β-glucan.

15. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 14, for use in a bispecific antigen-binding protein, antibody, or antigen-binding fragment thereof that binds two different antigens or epitopes.

16. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 15, for use in a multispecific antigen-binding protein, antibody, or antigen-binding fragment thereof that binds two or more different antigens or epitopes.

17. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 16, which is in the form of a homodimer or homotrimer.

18. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 17, having an EC50 of less than 0.2 μg / mL.

19. 1.5 × 10 －８ K lower than M Ｄ The antibody or antigen-binding fragment thereof according to any one of claims 1 to 18, having a value.

20. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 19, which has heat resistance up to 80°C.

21. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 20 for use as an antifungal agent.

22. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 21 for use in the treatment of fungal infections.

23. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 22 for use in the manufacture of a medicament for treating fungal infections.

24. A method for treating a fungal infection, comprising the step of administering an antibody or antigen-binding fragment thereof according to any one of claims 1 to 23.

25. An antibody or antigen-binding fragment thereof for use according to any one of claims 21 to 23, or the method according to claim 24, wherein the fungal infection is an Aspergillus infection.

26. An antibody or antigen-binding fragment thereof for use according to any one of claims 21 to 23, or the method according to claim 24, wherein the fungal infection is a Candida infection.

27. A recombinant fusion protein comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 26 and a peptide toxin.

28. (a) at least one of SEQ ID NO: 14 and SEQ ID NO: 16, (b) at least one of SEQ ID NO: 15 and SEQ ID NO: 17, (c) at least one of SEQ ID NO: 14 and SEQ ID NO: 18, (d) at least one of SEQ ID NO: 14 and SEQ ID NO: 19, (e) at least one of SEQ ID NO: 14 and SEQ ID NO: 20, (f) at least one of SEQ ID NO: 14 and SEQ ID NO: 17, or (g) at least one of SEQ ID NO: 14 and SEQ ID NO: 29 is linked to a peptide toxin, the recombinant fusion protein according to claim 27.

29. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 26 conjugated to a coupling moiety.

30. The antibody or antigen-binding fragment thereof according to claim 29, wherein the coupling moiety is a drug, toxin, cytokine, enzyme, or enzyme.

31. The antibody or antigen-binding fragment thereof according to claim 29 or 30, wherein the antibody or antigen-binding fragment and the coupling moiety are conjugated by a linker molecule.

32. The antibody or antigen-binding fragment thereof according to claim 31, wherein the linker molecule is a cleavable linker molecule.

33. A nucleic acid encoding the antibody or antigen-binding fragment thereof according to any one of claims 1 to 26 and 29 to 32.

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