Combination and use of microorganism and antibody

Abstract

The present disclosure relates to: a combination composition of microorganisms that are beneficial to an organism to be administered and an antibody that does not bind to microorganisms beneficial to the organism to be administered but binds to microorganisms that adversely affect the organism to be administered; and a method for using, in combination, microorganisms that are beneficial to the organism to be administered and an antibody that does not bind to microorganisms beneficial for the organism to be administered but binds to microorganisms that adversely affect the organism to be administered.

Description

Combinations and uses of microorganisms and antibodies

[0001] The present invention relates to a method and composition for using in combination microorganisms that are beneficial to the living body, such as fermentation bacteria, with antibodies that do not bind to the beneficial microorganisms and have the effect of improving the intestinal flora.

[0002] The intestinal tract is essentially "outside the body" for the living body, and is therefore constantly exposed to a large number of diverse intestinal bacteria, viruses, food-derived antigens, etc. These intestinal bacteria form the intestinal flora.

[0003] It has been revealed that this intestinal flora performs various functions for the body through the mucosal surface composed of intestinal epithelial cells, etc., and it is also known that without the presence of intestinal bacteria, the intestinal immune system cannot develop normally (Non-Patent Documents 1 to 3).

[0004] When the intestinal flora changes and the symbiotic relationship with the host is disrupted, the intestinal immune system is overstimulated, disrupting its homeostasis and ultimately inducing many diseases such as inflammatory bowel disease, colon cancer, asthma, allergies, obesity, etc. For these reasons, it is known that the intestinal immune system not only eliminates pathogens, but also plays an important role in maintaining the homeostasis of the entire immune system (Non-Patent Documents 4 to 7).

[0005] C. difficile, a bacterium associated with inflammatory bowel disease (IBD), is a bacterium that, along with MRSA, is a cause of hospital-acquired infections and requires caution. In fact, U.S. statistics indicate that approximately 400,000–500,000 people are infected annually, resulting in approximately 15,000–20,000 deaths. C. difficile infections often occur in patients receiving long-term antibiotic therapy, and once infected, the recurrence rate is high (15–35%). Therefore, among the bacteria associated with IBD, there is a particularly strong demand for effective treatments. While current treatments involve powerful antibiotics such as vancomycin, these methods leave residual bacterial toxins and promote the development of antibiotic-resistant bacteria, necessitating the development of alternative treatments.

[0006] Despite previous attempts, many patients still suffer from C. difficile enteritis and other inflammatory bowel diseases associated with intestinal microbiota. Furthermore, fecal transplants have been attempted as relatively new approaches, but their therapeutic effects are not consistent. The inventors of the present disclosure have developed antibodies and the like that bind to C. difficile bacteria and can be used for the treatment of inflammatory bowel diseases (Patent Document 1: WO2023 / 277142, Patent Document 2: WO2023 / 277166). However, further development of methods and pharmaceuticals for preventing or treating inflammatory bowel diseases associated with intestinal microbiota is desired.

[0007] WO2023 / 277142WO2023 / 277166

[0008] Cerf-Bensussan N, and Gaboriau-Routhiau V. , Nat Rev Immunol 2010;10:735 Hooper LV, and Macpherson AJ. , Nat Rev Immunol 2010;10:159Round JL, and Mazmanian SK, Proc Natl Acad Sci USA 2010;107:12204Vijay-Kumar M, et al. , Science 2010; 328:228 Shulzhenko N, et al. , Nat Med 2011;17:1585Bry L, et al. , Science 1996; 273:1380 Turnbaugh PJ, et al. , Nature 2006;444:1027

[0009] An objective of the present disclosure is to provide new compositions and methods for use in imparting or enhancing an intestinal flora improving effect, an immunostimulating effect, an anti-inflammatory effect, or an in vivo bactericidal effect.

[0010] The inventors of the present disclosure have discovered that by administering a combination of microorganisms that are beneficial to the target organism and antibodies that do not bind to the microorganisms that are beneficial to the target organism and have an intestinal flora-improving effect, excellent effects can be obtained in improving the gastrointestinal flora of the target organism, imparting and promoting immunostimulatory, anti-inflammatory, and bactericidal effects in the target organism, or in preventing or treating symptoms or diseases that require the imparting or promotion of these effects.

[0011] In one aspect, the present disclosure provides the following composition as a new composition for use in the prevention or treatment of symptoms or diseases that require the imparting or enhancement of an intestinal microbiota-improving effect, an immunostimulating effect, an anti-inflammatory effect, or an in vivo bactericidal effect, or that require the imparting or enhancement of these effects: A composition for use in the prevention or treatment of symptoms or diseases that require the imparting or enhancement of an intestinal microbiota-improving effect in an organism to which it is administered, characterized in that it comprises an antibody or an antigen-binding fragment thereof that has an intestinal microbiota-improving effect and does not bind to microorganisms that are beneficial to the organism to which it is administered, the antibody or antigen-binding fragment thereof having an intestinal microbiota-improving effect and that does not bind to microorganisms that are beneficial to the organism to which it is administered, characterized in that it is administered in combination with microorganisms that are beneficial to the organism to which it is administered.

[0012] In one aspect, the present disclosure provides the following composition as the new composition: A composition for use in improving the intestinal microbiota of a target organism, imparting or enhancing an immunostimulatory effect in a target organism, an anti-inflammatory effect in a target organism, or a microbicidal effect in a target organism, or in preventing or treating a symptom or disease requiring the imparting or enhancing of these effects, characterized in that the composition contains microorganisms beneficial to the target organism, and is administered in combination with an antibody or antigen-binding fragment thereof that does not bind to the microorganisms beneficial to the target organism and has an intestinal microbiota-improving effect.

[0013] In one aspect, the present disclosure provides the following method as a new method for use in the prevention or treatment of symptoms or diseases that require the imparting or enhancement of an intestinal flora-improving effect, an immunostimulating effect, an anti-inflammatory effect, or an in vivo bactericidal effect, or that require the imparting or enhancement of these effects: A method for preventing or treating symptoms or diseases that require the imparting or enhancement of an intestinal flora-improving effect, an immunostimulating effect in a target organism, an anti-inflammatory effect in a target organism, or a bactericidal effect in a target organism, comprising the step of administering to a subject to whom, is being administered, or is scheduled to be administered, a microorganism beneficial to the target organism, in need of the imparting or enhancement of an intestinal flora-improving effect, an immunostimulating effect in a target organism, an anti-inflammatory effect in a target organism, or a bactericidal effect in a target organism, an antibody that does not bind to the microorganism beneficial to the target organism.

[0014] In one aspect, the present disclosure provides the following method as the above-mentioned new method: A method for preventing or treating a symptom or disease requiring the imparting or enhancement of an intestinal flora-improving effect in a subject, an immunostimulatory effect in a subject, an anti-inflammatory effect in a subject, or a bactericidal effect in a subject, or a symptom or disease requiring the imparting or enhancement of these effects, comprising the step of administering to a subject to whom, or who is scheduled to be administered, a microorganism beneficial to the subject, an antibody that does not bind to the microorganism beneficial to the subject.

[0015] In one aspect, the present disclosure provides the following in more detail: [Item 1] A composition comprising an antibody or antigen-binding fragment thereof that does not bind to microorganisms beneficial to the target organism and has the effect of improving the intestinal flora, characterized in that it is administered in combination with microorganisms beneficial to the target organism. [Item 2] A composition comprising microorganisms beneficial to the target organism, characterized in that it is administered in combination with an antibody or antigen-binding fragment thereof that does not bind to microorganisms beneficial to the target organism. [Item 3] The composition of Item 1 or 2, wherein the microorganisms beneficial to the target organism are microorganisms selected from the group consisting of microorganisms that produce short-chain fatty acids, microorganisms that suppress the production of secondary bile acids, microorganisms that suppress the growth of microorganisms harmful to the target organism, and microorganisms that suppress the production of toxins from microorganisms that adversely affect the target organism. [Item 4] The composition of Item 1 or 2, wherein the microorganism beneficial to the recipient organism is Clostridium scindens, Lactobacillus paracasei, and / or Bifidobacterium breve. [Item 5] The composition of Item 1 or 2, wherein the antibody is an antibody that binds to a microorganism that adversely affects the recipient organism. [Item 6] The composition of Item 1 or 2, wherein the microorganism that adversely affects the recipient organism is a disease-associated microorganism or a toxin-producing microorganism. [Item 7] The composition of Item 1 or 2, wherein the microorganism that adversely affects the recipient organism is Escherichia coli or Clostridioides difficile. [Item 8] The antibody comprises: a) a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 8; b)an antibody comprising: a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 17; and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 18; c) an antibody comprising: a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 27; and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 28; d) a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 81; an antibody comprising a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 82; e) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 93, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 94; f) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 37, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 38; or g) For a reference antibody comprising a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 37 and a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 38,Item 9: The composition of Item 1 or 2, which is an antibody having at least one amino acid mutation in at least one region selected from heavy chain CDR1-3, light chain CDR1-3, and light chain FR1, and which binds to the amino acid sequence RQEEHIELIAS in Escherichia coli SHMT protein and the amino acid sequence VLDMMMKLEKPE in C. difficile iPGM protein. [Item 10] The composition of Item 1 or 2, which is a composition for use in improving the intestinal microflora, imparting or enhancing an immunostimulatory effect in a subject, an anti-inflammatory effect in a subject, or a microbicidal effect in a subject, or for use in preventing or treating a symptom or disease requiring the impartation or enhancement of these effects. [Item 11] The composition of Item 1 or 2, which is a composition for use in preventing or treating intestinal microbial symbiotic imbalance (dysbiosis), inflammatory bowel disease (IBD), or bacterial enteritis. [Item 11] The composition of Item 1 or 2, which is for use in the prevention or treatment of inflammatory bowel disease (IBD) or bacterial enteritis in a subject with an intestinal microbial dysbiosis imbalance. [Item 12] The composition of Item 1 or 2, which is further administered in combination with an oligosaccharide. [Item 13] The composition of Item 1 or 2, which comprises microorganisms beneficial to the subject and the antibody or antigen-binding fragment thereof in the same composition. [Item 14] The composition of Item 1 or 2, which comprises microorganisms beneficial to the subject and the antibody or antigen-binding fragment thereof in separate compositions. [Item 15] The composition of Item 1 or 2, wherein the antibody or antigen-binding fragment thereof is included in the composition as a nucleic acid or set of nucleic acids encoding them. [Item 16] A method for treating a subject, comprising the step of administering microorganisms beneficial to the subject and an antibody or antigen-binding fragment thereof that has an intestinal microbiota-improving effect and does not bind to the microorganisms beneficial to the subject. [Item 17] A method for treating a subject, comprising the step of administering an antibody or antigen-binding fragment thereof having an intestinal flora-improving effect to a subject who has been administered, is currently being administered, or is scheduled to be administered a microorganism beneficial to the subject. [Item 18]A method for treating a subject who has received, is receiving, or is scheduled to receive an antibody or antigen-binding fragment thereof having an intestinal microbiota-improving effect, comprising a step of administering microorganisms beneficial to the subject. [Item 19] The method of any one of Items 16 to 18, wherein the microorganisms beneficial to the subject are selected from the group consisting of microorganisms that produce short-chain fatty acids, microorganisms that suppress the production of secondary bile acids, microorganisms that suppress the growth of microorganisms harmful to the subject, and microorganisms that suppress the production of toxins from microorganisms that adversely affect the subject. [Item 20] The method of any one of Items 16 to 18, wherein the microorganisms beneficial to the subject are Clostridium sindens, Lactobacillus paracasei, and / or Bifidobacterium breve. [Item 21] The method according to any one of Items 16 to 18, wherein the antibody is an antibody that binds to a microorganism that adversely affects a subject to be administered. [Item 22] The method according to any one of Items 16 to 18, wherein the microorganism that adversely affects a subject to be administered is Escherichia coli or Clostridioides difficile. [Item 23] The antibody is: a) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 8; b) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 17, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 18; c) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 19;an antibody comprising: a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 27, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 28; d) an antibody comprising: a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 81, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 82; e) a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 93, and an antibody comprising a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO:94; f) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO:37, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO:38; or g) an antibody having at least one amino acid mutation in at least one region selected from heavy chain CDR1 to 3, light chain CDR1 to 3, and light chain FR1, relative to a reference antibody comprising a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO:37, and a light chain variable region comprising the amino acid sequence represented by SEQ ID NO:38, 24. The method according to any one of items 16 to 18, wherein the antibody binds to the amino acid sequence VLDMMMKLEKPE in the iPGM protein of C. difficile.The method according to any one of items 16 to 18, for preventing or treating a symptom or disease requiring the imparting or enhancement of an intestinal microbiota-improving effect, an immunostimulating effect, an anti-inflammatory effect, or a microbicidal effect, or for preventing or treating the symptom or disease in a subject having such a symptom or disease requiring the imparting or enhancement of such an effect. [Item 25] The method according to any one of items 16 to 18, for preventing or treating an intestinal microbial dysbiosis, an inflammatory bowel disease (IBD), or bacterial enteritis in a subject having such a symptom or disease. [Item 26] The method according to any one of items 16 to 18, for preventing or treating an intestinal microbial dysbiosis and inflammatory bowel disease (IBD), or an intestinal microbial dysbiosis and bacterial enteritis in a subject having such a symptom or disease. [Item 27] ​​The method of any one of Items 16 to 18, further comprising administering an oligosaccharide. [Item 28] The method of any one of Items 16 to 18, wherein the antibody or antigen-binding fragment thereof is administered as a nucleic acid or a set of nucleic acids encoding the antibody or antigen-binding fragment thereof.

[0016] The present disclosure has the effect of providing new compositions and methods for imparting or enhancing an intestinal flora-improving effect, an immunostimulating effect, an anti-inflammatory effect, or an in vivo bactericidal effect, or for use in the prevention or treatment of symptoms or diseases requiring the impartation or enhancement of these effects.

[0017] Figure 1 shows that the rW27 antibody having the intestinal flora-improving effect binds to Echerichia coli and Clostridioides difficile. Figure 2 shows that the rW27 antibody having the intestinal flora-improving effect binds to Clostridium butyricum and Clostridium saccharolyticum. Figure 3 shows that the rW27 antibody having the intestinal flora-improving effect does not bind to Clostridium sindens, Lactobacillus paracasei, and Bifidobacterium breve, which are microorganisms beneficial to the organism to which it is administered. Figure 4 shows that the rW27 antibody having the intestinal flora-improving effect binds to C. Figure 5 is a diagram showing the effect of combined administration of an rW27 antibody having an intestinal flora-improving effect and lactic acid bacteria on C. difficile infection, using body weight change as an index. Figure 6 is a diagram showing the effect of combined administration of an rW27 antibody having an intestinal flora-improving effect and lactic acid bacteria on C. difficile infection, using survival rate as an index. Figure 7 is a diagram showing changes in the intestinal flora composition when combined administration of an rW27 antibody having an intestinal flora-improving effect and lactic acid bacteria on C. difficile infection. 10 shows changes in the diversity of the intestinal flora when rW27 antibody, which has the effect of improving the intestinal flora, and lactic acid bacteria are administered in combination in C. difficile infection.

[0018] The inventors of the present disclosure have discovered that by administering a combination of microorganisms that are beneficial to the target organism and antibodies that do not bind to the microorganisms that are beneficial to the target organism and have an intestinal flora-improving effect, excellent effects can be obtained in improving the gastrointestinal flora of the target organism, imparting and promoting immunostimulatory, anti-inflammatory, and bactericidal effects in the target organism, or in preventing or treating symptoms or diseases that require the imparting or promotion of these effects.

[0019] More specifically, they found that the combined administration of microorganisms beneficial to the target organism, such as lactic acid bacteria, and IgA antibodies having the effect of improving the intestinal flora, provides a qualitatively different and superior infection prevention effect compared to the administration of either microorganism alone. That is, in the case of C. difficile infection in a state of intestinal bacterial symbiotic imbalance, the single administration of microorganisms beneficial to the target organism, such as lactic acid bacteria, induces symptoms such as diarrhea, whereas the combined administration of an extremely low dose of IgA antibodies having the effect of improving the intestinal flora with the microorganism suppresses the diarrheal symptoms, enabling effective suppression of infection-related symptoms such as diarrhea at an earlier stage.

[0020] The present disclosure provides a composition or method that provides synergistically excellent effects in improving the gastrointestinal microbiota of a target organism, imparting and promoting immunostimulatory, anti-inflammatory, and bactericidal effects in the target organism, or preventing or treating symptoms or diseases that require the imparting or promotion of these effects, by administering a combination of microorganisms that are beneficial to the target organism and an antibody that has an intestinal microbiota-improving effect but does not bind to the microorganisms that are beneficial to the target organism.

[0021] (Definitions) In this specification, when multiple ranges of numerical values ​​are given, the same applies to ranges formed by any combination of the lower and upper limits of those multiple ranges.

[0022] As used herein, the term "antibody" is used in the broadest sense and includes, but is not limited to, monoclonal antibodies, polyclonal antibodies, and antibody fragments that exhibit the intended antigen-binding activity. A full-length antibody comprises a heavy chain and a light chain that are primarily composed of polypeptides. The heavy chain and light chain each contain a region called a variable region that recognizes an antigen, and these regions are generally referred to as the heavy chain variable region and the light chain variable region, respectively. The variable regions each contain CDRs 1 to 3, which are further specified as antigen-recognition sites, in order from the amino terminus. These CDRs 1 to 3 are also more specifically referred to as heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, light chain CDR3, etc. Furthermore, the regions other than CDRs 1 to 3 of the heavy chain and light chain are referred to as heavy chain FRs 1 to 4 and light chain FRs 1 to 4, respectively, in order from the amino terminus. The antibody may be in a form consisting of two heavy chains and two light chains, or in a form consisting of one heavy chain and one light chain (also called a single-chain antibody).

[0023] The antibody may be in the form of an antibody consisting of two heavy chains and two light chains, or in the form of an antibody consisting of one heavy chain and one light chain (also called a single-chain antibody).Furthermore, the antibody may be in the form of a dimer having a J (joining) chain and, in addition to the two heavy chains and two light chains, and optionally an SC (secretary component), or in the form of a multimer such as a trimer, tetramer, or pentamer.

[0024] There are various classes of antibodies, such as IgG, IgE, IgM, IgD, IgA, and IgY, and further subclasses thereof, such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0025] The antibodies of the present disclosure may have amino acid sequences derived from the same species or from different species, such as human, mouse, rat, hamster, rabbit, goat, donkey, pig, cow, horse, chicken, monkey, chimpanzee, camel, and llama.

[0026] In the case of antibodies derived from different species, there is no particular limitation, and examples include antibodies derived from two or more of human, mouse, rat, hamster, rabbit, goat, sheep, donkey, pig, cow, horse, chicken, monkey, chimpanzee, camel, llama, etc.

[0027] As used herein, the term "antigen-binding fragment" of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been found that the ability of an antibody to specifically bind to an antigen can also be maintained by fragments consisting of a portion thereof. In one embodiment, the "antigen-binding fragment" of an antibody may be, but is not limited to, a Fab fragment consisting of a light chain variable region (VL), a heavy chain variable region (VH), a light chain constant region (CL), and a CH1 domain that is part of the heavy chain constant region; an F(ab')2 fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of a single antibody arm; a dAb fragment containing a single variable domain; and an isolated complementarity-determining region (CDR).

[0028] The antibody of the present disclosure may be a CDR-grafted antibody. In one example, a CDR-grafted antibody has part or all of the CDR region sequence of an antibody derived from one animal species replaced with the CDR sequence of another animal species. For example, one or more CDRs of a mouse antibody are replaced with the CDR sequence of a human antibody.

[0029] Methods well known to those skilled in the art can be used to identify heavy chain CDR1 to 3 in the heavy chain variable region and light chain CDR1 to 3 in the light chain variable region of an antibody. For example, the "Kabat definition" (Kabat et al., Ann. NY Acad. Sci. 1971, Vol. 190, pp. 382-391 and Kabat, E.A. et al., Sequences of Proteins of Immunological Interest, 5th ed., 1991, U.S. Department of Health and Human Services, NIH Publication, pp. 91-3242), the "Chothia definition" (Chothia et al., Nature, 1989, Vol. 342, pp. 877-883), and the "contact definition" (MacCallum et al., Nature, 1989, Vol. 342, pp. 877-883), all of which are well known to those skilled in the art, may be used. al., J. Mol. Biol., 1996, Vol. 262, pp. 732-745) can be used. To identify the CDR sequences in an antibody, the CDRs may be identified based on information in public databases (e.g., https: / / www.ncbi.nlm.nih.gov / igblast).

[0030] As used herein, "identity" refers to the degree to which two or more comparable amino acid sequences or nucleotide sequences are identical to each other. Thus, the higher the identity between two amino acid sequences or nucleotide sequences, the higher the identity or similarity between those sequences. The level of identity between amino acid sequences or nucleotide sequences is usually determined using the sequence analysis tool FASTA with default parameters. Alternatively, it can be determined using the BLAST algorithm by Karlin and Altschul (e.g., Karlin S, Altschul S F. Proc. Natl Acad Sci USA. 87:2264-2268 (1990), Karlin S, Altschul S F. Natl Acad Sci USA. 90:5873-7 (1993), etc.). Programs called BLASTN and BLASTX based on such BLAST algorithms have been developed (e.g., Altschul S F, Gish W, Miller W, Myers E W, Lipman D J. J Mol Biol. 215:403-10 (1990)). Specific techniques for these analysis methods are known, and can be found on the NCBI website. For example, when a certain amino acid sequence A is identical to another amino acid sequence B by a certain percentage, this means that amino acid sequence A and amino acid sequence B have the specified percentage of identity.

[0031] As used herein, "monoclonal" is a modifier indicating the character of an antibody or the like obtained from a population of substantially homogeneous antibodies, the individual antibodies within which are identical except for possible naturally occurring mutations that may be present in minor amounts.

[0032] As used herein, the term "conservative substitution techniques" refers to techniques in which an amino acid residue is replaced with an amino acid residue having a similar side chain.

[0033] For example, conservative substitution techniques include substitutions between amino acid residues having basic side chains such as lysine, arginine, and histidine. Other conservative substitution techniques include substitutions between amino acid residues having acidic side chains such as aspartic acid and glutamic acid; amino acid residues having uncharged polar side chains such as glycine, asparagine, glutamine, serine, threonine, tyrosine, and cysteine; amino acid residues having nonpolar side chains such as alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; amino acid residues having β-branched side chains such as threonine, valine, and isoleucine; and amino acid residues having aromatic side chains such as tyrosine, phenylalanine, tryptophan, and histidine.

[0034] As used herein, "prevention" of a condition, disorder, or disease has its usual meaning in the art, such as preventing the occurrence of a condition, disorder, or disease, reducing the risk of the occurrence of a condition, disorder, or disease, etc.

[0035] As used herein, "treatment" of a condition, disorder, or disease has its usual meaning in the art, and means, for example, preventing the onset of a condition, disorder, or disease; reducing the risk of onset of a condition, disorder, or disease; inhibiting the progression of a condition, disorder, or disease that has already occurred; delaying the progression of a condition, disorder, or disease that has already occurred; eliminating a condition, disorder, or disease that has already occurred; ameliorating the symptoms of a condition, disorder, or disease that has already occurred; and the like.

[0036] 1. Combination Composition of Microorganisms Beneficial to the Target Organism and an Antibody Having an Intestinal Microbiota-Improvement Activity That Does Not Bind to the Microorganisms Beneficial to the Target Organism In one aspect, the present disclosure provides a combination composition of Microorganisms Beneficial to the Target Organism and an antibody or antigen-binding fragment thereof having an intestinal microbiota-improving activity that does not bind to the microorganisms beneficial to the target organism. In this way, by using a combination of Microorganisms Beneficial to the Target Organism and an antibody or antigen-binding fragment thereof having an intestinal microbiota-improving activity that does not bind to the microorganisms beneficial to the target organism, it is possible to obtain synergistically excellent effects in improving the gastrointestinal microbiota of the target organism, imparting and promoting immunostimulatory, anti-inflammatory, and bactericidal effects in the target organism, or in preventing or treating symptoms or diseases that require the imparting or promotion of these effects.

[0037] In one aspect, the present disclosure provides the following composition: The composition comprises an antibody or antigen-binding fragment thereof that has an intestinal microbiota-improving effect and does not bind to microbes that are beneficial to the organism to which it is administered, characterized in that the antibody or antigen-binding fragment thereof is administered in combination with microbes that are beneficial to the organism to which it is administered, and that has an intestinal microbiota-improving effect, and that does not bind to microbes that are beneficial to the organism to which it is administered, for use in improving the intestinal microbiota of the organism to which it is administered, imparting or enhancing an immunostimulatory effect in the organism to which it is administered, an anti-inflammatory effect in the organism to which it is administered, or a microbicidal effect in the organism to which it is administered, or in preventing or treating a symptom or disease that requires the imparting or enhancing of these effects.

[0038] In one aspect, it has been experimentally confirmed that the antibody or its antigen-binding fragment does not bind to microorganisms beneficial to the target organism. The binding of the antibody or its antigen-binding fragment to the microorganism can be achieved by methods known to those skilled in the art. For example, the lack of binding of the antibody or its antigen-binding fragment to the microorganism can be confirmed by an ELISA test or the like.

[0039] The microorganisms that are beneficial to the organism to be administered in combination with the composition of the present disclosure may be contained in the same composition as the antibody, or may be contained in a separate composition.

[0040] In another aspect, the present disclosure provides the following composition: The composition comprises a microorganism beneficial to the organism to which it is administered, and is administered in combination with an antibody or antigen-binding fragment thereof that does not bind to the microorganism beneficial to the organism to which it is administered and has an intestinal microbiota-improving effect, and that has an intestinal microbiota-improving effect, and that does not bind to the microorganism beneficial to the organism to which it is administered, for use in improving the intestinal microbiota of the organism to which it is administered, imparting or enhancing an immunostimulatory effect in the organism to which it is administered, an anti-inflammatory effect in the organism to which it is administered, or a microbicidal effect in the organism to which it is administered, or in preventing or treating a symptom or disease requiring the imparting or enhancing of these effects.

[0041] In one aspect, it has been experimentally confirmed that the antibody or its antigen-binding fragment does not bind to microorganisms beneficial to the target organism. The binding of the antibody or its antigen-binding fragment to the microorganism can be achieved by methods known to those skilled in the art. For example, the lack of binding of the antibody or its antigen-binding fragment to the microorganism can be confirmed by an ELISA test or the like.

[0042] The antibody may be contained in the same composition as the microorganism that is beneficial to the organism to which it is administered, or may be contained in a separate composition.

[0043] (Microorganisms Beneficial to the Organism to Which the Antibody of the Present Disclosure is Administered) Microorganisms beneficial to the organism to which the antibody of the present disclosure is administered in a combined composition containing a microorganism beneficial to the organism to which the antibody is administered are not particularly limited, and include microorganisms that have a beneficial effect on the survival, health, etc. of the organism to which the antibody is administered. In one embodiment, the microorganisms beneficial to the organism to which the antibody is administered may be lactic acid bacteria, bifidobacteria, butyric acid bacteria, saccharifying bacteria, and the like, which are generally recognized as probiotics. In one embodiment, the microorganisms beneficial to the organism to which the antibody is administered may be microorganisms that produce metabolites beneficial to the organism to which the antibody is administered. In one embodiment, the microorganisms beneficial to the organism to which the antibody is administered may be microorganisms that produce short-chain fatty acids, for example, propionic acid.

[0044] In one embodiment, the microorganisms beneficial to the organism to which the administration is directed are microorganisms that maintain or improve the intestinal flora. In one embodiment, the microorganisms beneficial to the organism to which the administration is directed are microorganisms that maintain or improve the diversity of the intestinal flora.

[0045] In one embodiment, the microorganisms beneficial to the target organism may be microorganisms that suppress the production of harmful substances in the target organism. In one embodiment, the microorganisms beneficial to the target organism may be microorganisms that suppress the production of secondary bile acids, microorganisms that suppress the growth of microorganisms harmful to the target organism, or microorganisms that suppress toxin production in microorganisms that adversely affect the target organism. In one embodiment, the microorganisms beneficial to the target organism suppress toxin production in microorganisms that adversely affect the target organism by secreting amino acids.

[0046] In one aspect, the microorganisms beneficial to the target organism are Absiella innocuum, Acidaminococcus fermentans, Acidaminococcus intestine, Agathobacter rectal, Agathobaculum sp、Akkkermansia muciniphila、Alistipes putredinis、Alistipes shahii、Alistipes sp.、Anaerostipes caccae、Anaerostipes caccae DSM 14662、Aerostipes hadrus、Aerotruncus colihominis、Anaerotruncus colihominis DSM 17241、Clostridium cluster IV bacteria、Clostridium cluster Bacteroidetes caccae, Bacteroides cellulosiliticus, Bacteroides clarus, Bacteroides dōrei、Bacteroides eggerthii、Bacteroides faecis、Bacteroides ffragilis、Bacteroides ovatus、Bacteroides salyersiae、Bacteroides sp.、Bacteroides sp. MANG, Bacteroides thetaiomicron, Bacteroides uniformis, Bacteroides xylanisolvens, Bacteroides_B vulgatus, Barnesiella intestini hominis, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium pseudocatenulatum、Bff､obacteriumm pseudolongum subsp.、Bilophila wadsworthia、Blautia coccoides、Blue faeces, Blue hansen, Blue lyme, Blue products, Blue spp. canine oral taxon bacterium、Clostridiales bacterium 1_7_47FAA、Clostridium、Clostridium sp. MLG055, Clostridium asparagiforme, Clostridium asparagiforme DSM 15981, Clostridium bacterium, Clostridium boltea, Clostridium boltea ATCC BAA-613, Clostridium citroniae, Clostridium disporicum, Clostridium hathawayi, Clostridium indolis, Clostridium innocuum, Clostridium lactatifermentans, Clostridium lavalense, Clostridium orbicindens, Clostridium ramosum, Clostridium ramosum JCM1298, Clostridium saccharolyticum, Clostridium saccharogumia, Clostridium saccharolyticum, Clostridium saccharolyticum WM1、Clostridium scindens、Clostridium sp.、Clostridium sp. 14616、Clostridium sp. 31600020000000000000、Clostridium symbiosum、Clostridium symbiosum WL-141163、Clostridium_Coprobacillus cateniformis, Coprococci_B comes, Dorea formicigenans, Dorea longicatena, Dracourtella massiliensis, Eisenbergiella tayi, Erysipelatoclostridium ramosum, Erysipelotrichaceae bacterium, Erysipelotrichaceae bacterium 2_2_44A, Escherichia coli、Eubacterium contortum、Eubacterium desmolans、Eubacterium fissicatena、Eubacterium limosum、Eubacterium rectale、Eubacterium_E hallii、Flavinofractor plautii、Fusicatenibacter saccharivorans、Fusobacterium ulcerans、Fusobacterium_A sp、Hungatella hathewâym、Lachos0.raceae bacteriumm、Lachnoespiraceae bacteriumm 3_1_57FAA_CT1、Lachnospiraceae bacterium A4、Lachnospiraraceae bacteria DJF_VP30、Lachnospiraceae bacterium HGA0140、Lachnospiraceae casei、Lactobacillus mucosae、Marvinbryanta formatexigens、Megasphaera elsdenii、Odoribacter splanchnicus、Orbiscindens、Oscillibacter valericigenes、Oscillospiracea bacterium、Oscillospiracea bacterium 061048、Paeniclostridium sordellii, Parabacteroides distasonis, Parabacteroides goldsteinii, Parabacteroides gordonii, Parabacteroides johnsonii, Parabacteroides merdae, Paraclostridium massiliensis, Paraprevotella xylaniphila, Parasuterella excrementinihominis, Phascolarctobacterium faecium, Prevotella Copri, Pseudoflavinofractor capillosus, Pseudoflavonifractor capillosus ATCC 29799, Roseburia faecis, Ruminococcus albus, Ruminococcus champanellensis, Ruminococcus faecis, Ruminococcus gnavus, Ruminococcus obeum, Ruminococcus sp. , Ruminococcus sp. 16442, Ruminococcus sp. ID8, Ruminococcus sp. M-1, Ruminococcus torques, Sellimonas intestinalis, Subdoligranulum sp., Subdolinogranulum spp., and Turicibacter sanguinis, and microorganisms containing a 16sRNA comprising a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence of the 16sRNA of these microorganisms. In particular, Clostridium scindens, Lactobacillus paracasei and / or Bifidobacterium breve, as well as nucleic acid sequences of the 16sRNA of these microorganisms, and nucleic acid sequences of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%,Preferably, the 16sRNA is selected from the group consisting of microorganisms containing a 16sRNA comprising a nucleic acid sequence having 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity.

[0047] Microorganisms that are beneficial to the target organism can have multiple effects on the target organism, and for example, depending on the condition of the target organism, they may have a beneficial effect on the survival, health, etc. of the target organism, or they may have a negative effect depending on the condition of the target organism. Such microorganisms can also be used as beneficial microorganisms for the target organism depending on the condition of the target organism.

[0048] (Antibody having an effect of improving intestinal flora) The antibody or antigen-binding fragment thereof having an effect of improving intestinal flora used in the combination composition of the present disclosure is not particularly limited, as long as it is an antibody or antigen-binding fragment thereof that does not bind to microorganisms that are beneficial to the target organism to be used in combination and has an effect of improving intestinal flora.

[0049] In the present disclosure, "an antibody or antigen-binding fragment thereof does not bind to a microorganism" means that the antibody or antigen-binding fragment thereof does not substantially bind to the microorganism in vivo. "Does not substantially bind" means that the antibody or antigen-binding fragment thereof does not bind to a level that significantly affects the survival, growth, or physiological function of the microorganism in vivo. In one aspect, "an antibody or antigen-binding fragment thereof does not bind to a microorganism" means that, when subjected to a standard ELISA test, the antibody exhibits less than twice the binding affinity (absorbance) of a control antibody at any of the following concentrations: 0.2 to 20 μg / mL, 0.3 to 15 μg / mL, 0.3 to 15 μg / mL, 0.4 to 12 μg / mL, 0.5 to 10 μg / mL, 0.6 to 8 μg / mL, 0.7 to 7 μg / mL, 0.8 to 6 μg / mL, 0.9 to 5 μg / mL, or 1 to 2 μg / mL. In one aspect, an antibody or antigen-binding fragment thereof of the present disclosure does not bind to a microorganism means that the dissociation constant (KD) for the microorganism is at least 10 times, preferably at least 20 times, more preferably at least 50 times, and even more preferably 100 times, 200 times, 500 times, or 1000 times higher than the dissociation constant (KD) of an antibody that binds to the microorganism.

[0050] In one aspect, an antibody having an intestinal microbiota-improving effect used in the combination of the present disclosure increases the diversity of the microbiota in the gastrointestinal contents or excrement of a subject organism to which it is administered. In one aspect, an antibody having an intestinal microbiota-improving effect used in the combination of the present disclosure includes an antibody that increases the amount of short-chain fatty acids in the gastrointestinal contents or excrement of a subject organism to which it is administered. In one aspect, an antibody having an intestinal microbiota-improving effect used in the combination of the present disclosure includes an antibody that binds to microorganisms that have a negative effect on the subject organism to which it is administered.

[0051] Microorganisms that adversely affect a target organism can have multiple effects on the target organism, and for example, depending on the condition of the target organism, they may have a beneficial effect on the survival, health, etc. of the target organism, or they may have an adverse effect depending on the condition of the target organism. The antibodies disclosed herein that have an intestinal microbiota-improving effect can also bind to such microorganisms.

[0052] (Microorganisms that have an adverse effect on a target organism) In the present disclosure, microorganisms that have an adverse effect on a target organism are not particularly limited and include microorganisms that have an adverse effect on the survival, health, etc. of the target organism. In one embodiment, the microorganisms that have an adverse effect on a target organism are microorganisms associated with a disease. In one embodiment, the microorganisms that have an adverse effect on a target organism are microorganisms that produce toxins. In one embodiment, the microorganisms that have an adverse effect on a target organism are microorganisms that reduce the diversity of the intestinal flora. In one embodiment, the microorganisms that have an adverse effect on a target organism may be selected from the group consisting of Escherichia coli and Clostridioides difficile, and microorganisms that contain a 16sRNA that contains a nucleic acid sequence that has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the nucleic acid sequence of the 16sRNA of these microorganisms.

[0053] The antibody or antigen-binding fragment thereof of the present disclosure may bind to fragments of microorganisms present in the subject upon their death, as long as it does not bind to microorganisms that are beneficial to the subject organism and does not inhibit their survival and / or growth.

[0054] When an antibody or antigen-binding fragment thereof of the present disclosure binds to a microorganism that adversely affects a target organism, the antibody or antigen-binding fragment thereof can bind to the whole microorganism or a fragment thereof. In one aspect, the antibody or antigen-binding fragment thereof of the present disclosure binds to a microorganism that adversely affects a target organism and has an effect of improving the intestinal flora. In one aspect, the antibody or antigen-binding fragment thereof binds to a microorganism that adversely affects a target organism and suppresses the survival, growth, toxin production, etc. of the microorganism, thereby having an effect of improving the intestinal flora.

[0055] In the present disclosure, "an antibody or antigen-binding fragment thereof binds to a microorganism" typically means that the antibody or antigen-binding fragment thereof binds to the microorganism in vivo and affects the survival, function, or growth of the microorganism. In one aspect, "an antibody or antigen-binding fragment thereof binds to a microorganism" means that, when subjected to a standard ELISA test, the antibody exhibits a binding affinity (absorbance) that is at least twice that of a control antibody at any of the following concentrations: 0.2 to 20 μg / mL, 0.3 to 15 μg / mL, 0.3 to 15 μg / mL, 0.4 to 12 μg / mL, 0.5 to 10 μg / mL, 0.6 to 8 μg / mL, 0.7 to 7 μg / mL, 0.8 to 6 μg / mL, 0.9 to 5 μg / mL, or 1 to 2 μg / mL.

[0056] In one aspect, the class and species of the antibody of the present disclosure are not particularly limited, and the configuration of the antigen-binding fragment is also not limited. Non-limiting specific examples of the antibody of the present disclosure are shown below.

[0057] Antibodies used in the combinations of the present disclosure include, for example, the antibodies and variants thereof described in WO2023 / 277142 and Japanese Patent Application No. 2023-131537. These antibodies bind to Escherichia coli and / or Clostridioides difficile that adversely affect the subject to which they are administered.

[0058] The antibodies used in the combinations of the present disclosure may be produced from antibody-producing cells derived from B cells, such as hybridomas, or may be produced by using genetic recombination technology to introduce nucleic acid encoding the antibody into cells outside the immune system, and used as recombinant antibodies.

[0059] In one aspect, the antibody, or antigen-binding fragment thereof, used in the combination of the present disclosure is the following antibody disclosed in WO2023 / 277142: an antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3, the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3, the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8.

[0060] In one aspect, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 1; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 2; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 3; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 4; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 5; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 6.

[0061] In one embodiment, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

[0062] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is a purified recombinant antibody SNK0001AR or an antigen-binding fragment thereof, which is an IgA antibody obtained by determining the nucleic acid sequence encoding the IgM antibody SNK0001M produced by a hybridoma obtained from spleen-derived B cells and applying recombinant technology. The antibody SNK0001AR has the following amino acid sequence:

[0063] In one aspect, an antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 17, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 18.

[0064] In one aspect, an antibody, or antigen-binding fragment thereof, used in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 11; a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 12; and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 13; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 14; a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 15; and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 16.

[0065] In one embodiment, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 18, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

[0066] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is a purified recombinant antibody SNK0002AR, which is an IgA antibody produced by determining the nucleic acid sequence encoding the IgM antibody SNK0002M produced by a hybridoma obtained from spleen-derived B cells and applying recombinant technology. The antibody SNK0002AR has the following amino acid sequence:

[0067] In one aspect, an antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:27, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:28.

[0068] In one aspect, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:21; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:22; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:23; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:24; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:25; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:26.

[0069] In one embodiment, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:27, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:28, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

[0070] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is the purified recombinant antibody SNK0003A or an antigen-binding fragment thereof, which is an IgA antibody obtained by applying recombinant technology to the IgA antibody SNK0003M produced by a hybridoma obtained from B cells derived from the intestinal lamina propria mucosa. The antibody SNK0003A has the following amino acid sequence:

[0071] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is described in Japanese Patent Application No. 2023-131537: an IgA antibody or antigen-binding fragment thereof comprising: a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 81; and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 82.

[0072] In one aspect, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 83; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 84; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 85; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 86; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 87; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 88.

[0073] In one embodiment, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:81, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:82, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

[0074] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 82, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 83; a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 84; and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 85; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 86; a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 87; and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 88.

[0075] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is SNK0004, which is an IgA antibody produced by a hybridoma obtained from mouse B cells, and the nucleic acid sequence encoding the antibody was determined and then converted into an IgA antibody using recombinant technology, or an antigen-binding fragment thereof. Antibody SNK0004 has the following amino acid sequence:

[0076] In one embodiment, the antibody SNK0004 is encoded by the following nucleic acid sequence:

[0077] In one embodiment, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is described in Japanese Patent Application No. 2023-131537: an IgA antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 93, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 94.

[0078] In one aspect, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:95; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:96; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:97; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:98; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:99; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:100.

[0079] In one embodiment, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:93, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:94, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

[0080] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:93, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:94, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and the antibody or antigen-binding fragment thereof comprises: a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO:95; a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO:96; and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO:97; and a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO:98; a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO:99; and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO:100.

[0081] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure is SNK0005, which is an IgA antibody produced by a hybridoma isolated from B cells derived from mouse small intestine, and the nucleic acid sequence encoding the antibody was determined and then recombinantly converted into an IgA antibody using recombinant technology. The antibody SNK0005 has the following amino acid sequence:

[0082] In one embodiment, the antibody SNK0005 is encoded by the following nucleic acid sequence:

[0083] In one aspect, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 31; a heavy chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 32; and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 33; and a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 34; a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 35; and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 36.

[0084] In one embodiment, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises: a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:37, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto; and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:38, or a sequence having at least 90%, at least 95%, at least 98%, or at least 99% sequence identity thereto.

[0085] In one aspect, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure has at least one amino acid mutation in at least one region selected from heavy chain CDR1-3, light chain CDR1-3, and light chain FR1, relative to a reference antibody comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 37 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 38, and is an antibody or antigen-binding fragment thereof that binds to the amino acid sequence RQEEHIELIAS (SEQ ID NO: 103) in the Escherichia coli SHMT protein and the amino acid sequence VLDMMMKLEKPE (SEQ ID NO: 104) in the iPGM protein of C. difficile.

[0086] The at least one amino acid mutation in the antibody can be identified by applying techniques available to those skilled in the art, utilizing the results of structural analysis of the complexes of an antibody having the amino acid sequence of the reference antibody with the E. coli SHMT protein, and an antibody having the amino acid sequence of the reference antibody with the C. difficile iPGM protein. The number of amino acid mutations relative to the reference antibody may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more.

[0087] Such a reference antibody is not particularly limited as long as it has a heavy chain variable region of W27G2_HV (SEQ ID NO: 37) and a light chain variable region of W27G2_LV (SEQ ID NO: 38). For example, an IgG antibody (W27RS_H000_L000GR) can be used. The antibody W27RS_H000_L000GR has the same heavy chain variable region and light chain variable region as the antibody W27G2 produced by a hybridoma obtained from B cells derived from the intestinal lamina propria mucosa.

[0088] Whether an antibody having the specified mutation actually binds to the amino acid sequence RQEEHIELIAS in the E. coli SHMT protein and the amino acid sequence VLDMMMKLEKPE in the iPGM protein of C. difficile can be confirmed by methods well known to those skilled in the art, such as ELISA or Western blot. For example, the test method disclosed in WO2023 / 277142 can be used.

[0089] In one embodiment, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises １ heavy chain CDR1 comprising the amino acid sequence of YYIH, RIDPENX ２ X ３ a heavy chain CDR2 comprising the amino acid sequence of TTYAPKFQ; a heavy chain CDR3 comprising the amino acid sequence of YCARSTVL; and a heavy chain variable region comprising: RX ４ a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence of SQSIVHTNG; a light chain CDR2 comprising the amino acid sequence of KLLIYKV; and a light chain CDR3 comprising the amino acid sequence of GVYYCFQGS; and a light chain FR1 comprising the amino acid sequence of TPLSLPVSLGDQA or SPASX. ５ SVSLGDRX ６ X １ , X ２ , X ３ are each independently a neutral polar amino acid or an acidic polar amino acid, and X ４ is a nonpolar amino acid or a neutral polar amino acid, and X ５ , X ６is an antibody or antigen-binding fragment thereof, each independently a nonpolar amino acid.

[0090] The above antibody was designed, based on three-dimensional structural analysis, to have a binding mode similar to that of the W27G2 antibody with respect to the amino acid sequence RQEEHIELIAS in the E. coli SHMT protein and the amino acid sequence VLDMMMKLEKPE in the iPGM protein of C. difficile, and actually exhibits a binding mode and bacterial growth inhibitory effect similar to that of the W27G2 antibody with respect to these bacteria.

[0091] In one embodiment, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises １ heavy chain CDR1 comprising the amino acid sequence of YYIH, RIDPENX ２ X ３ a heavy chain CDR2 comprising the amino acid sequence of TTYAPKFQ; a heavy chain CDR3 comprising the amino acid sequence of YCARSTVL; and a heavy chain variable region comprising: RX ４ a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence of SQSIVHTNG; a light chain CDR2 comprising the amino acid sequence of KLLIYKV; and a light chain CDR3 comprising the amino acid sequence of GVYYCFQGS; and a light chain FR1 comprising the amino acid sequence of TPLSLPVSLGDQA or SPASX. ５ SVSLGDRX ６ X １ , X ２ are each independently asparagine or aspartic acid, and X ３ is glutamine or glutamic acid, and X ４ is alanine or serine, and X ５ is leucine or methionine, and X ６ is an alanine or valine.

[0092] In one embodiment, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 31 or 41; a heavy chain CDR2 comprising the amino acid sequence represented by any one of SEQ ID NOs: 32, 42-44; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 33.

[0093] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 31; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 32; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 33.

[0094] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:31; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:42; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:33.

[0095] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:31; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:43; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:33.

[0096] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 31; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 44; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 33.

[0097] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:41; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:32; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:33.

[0098] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:41; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:42; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:33.

[0099] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:41; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:43; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:33.

[0100] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a heavy chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:41; a heavy chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:44; and a heavy chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:33.

[0101] In one aspect, the heavy chain of an antibody used in the combination of the present disclosure comprises: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 34 or 52; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 35; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 36.

[0102] In one aspect, the light chain of an antibody used in the combination of the present disclosure comprises: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO: 34; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO: 35; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO: 36.

[0103] In one aspect, the light chain of an antibody used in the combination of the present disclosure comprises: a light chain CDR1 comprising the amino acid sequence represented by SEQ ID NO:52; a light chain CDR2 comprising the amino acid sequence represented by SEQ ID NO:35; and a light chain CDR3 comprising the amino acid sequence represented by SEQ ID NO:36.

[0104] In one embodiment, an antibody or antigen-binding fragment thereof used in the combination of the present disclosure comprises, in its light chain variable region, a sequence that binds to Protein L. An antibody molecule or antigen-binding fragment thereof that comprises the sequence that binds to Protein L can be purified using a Protein L column.

[0105] In one aspect, the antibody or antigen-binding fragment thereof comprising a sequence that binds to Protein L of the present disclosure has the amino acid sequence SPASX ５ SVSLGDRX ６ a light chain variable region comprising X ５ , X ６ are each independently a nonpolar amino acid.

[0106] In one aspect, the antibody or antigen-binding fragment thereof comprising a sequence that binds to Protein L of the present disclosure has the amino acid sequence SPASX ５ SVSLGDRX ６ a light chain variable region comprising X ５ is leucine or methionine, and X ６ is alanine or valine.

[0107] In one embodiment, the antibody or antigen-binding fragment thereof comprising a sequence that binds to Protein L of the present disclosure comprises a light chain variable region comprising an amino acid sequence represented by one selected from the group consisting of SEQ ID NOs: 53-55.

[0108] In one aspect, the antibody or antigen-binding fragment thereof comprising a sequence that binds to Protein L of the present disclosure comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 54. This sequence is obtained by introducing a mutation into the sequence TPLSLPVSLGDQA (SEQ ID NO: 56) of the light chain FR1 region of the W27G2 antibody so that the antibody molecule binds to Protein L.

[0109] In one aspect, the antibody or antigen-binding fragment thereof comprising a sequence that binds to Protein L of the present disclosure comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 55. This sequence is obtained by further introducing a mutation into the light chain FR1 region of the light chain variable region comprising the amino acid sequence of SEQ ID NO: 54, so that the antibody molecule binds to Protein L more tightly.

[0110] In one aspect, the antibody or antigen-binding fragment thereof comprising a sequence that binds to Protein L of the present disclosure comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 58. This sequence is obtained by further introducing a mutation into the light chain FR1 region of the light chain variable region comprising the amino acid sequence of SEQ ID NO: 54, so that the antibody molecule binds to Protein L more tightly.

[0111] In one aspect, the antibodies or antigen-binding fragments thereof used in the combinations of the present disclosure may have any combination of heavy and light chains described above, and may or may not contain a sequence that binds to Protein L.

[0112] In one aspect, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure comprises a heavy chain variable region comprising: a heavy chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 31 or 41; a heavy chain CDR2 comprising the amino acid sequence set forth in any of SEQ ID NOs: 32, 42-44; and a heavy chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 33; a light chain variable region comprising: a light chain CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 34 or 52; a light chain CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 35; and a light chain CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 36; and a light chain FR1 comprising the amino acid sequence set forth in any of SEQ ID NOs: 53-56.

[0113] For example, an antibody or antigen-binding fragment thereof for use in the combination of the present disclosure is an antibody or antigen-binding fragment thereof that comprises a combination of any of the heavy chain variable regions and any of the light chain variable regions described below.

[0114] In one embodiment, the IgA antibody or antigen-binding fragment thereof preferably comprised in the composition of the present disclosure comprises the following combination of heavy chain variable regions and light chain variable regions:

[0115] In one aspect, the antibodies or antigen-binding fragments thereof used in the combinations of the present disclosure include the antibodies described below or their IgA variants.

[0116] Antibodies produced in CHO cells by introducing mutations into the heavy or light chain of the W27G2 antibody so as not to substantially alter its antigen specificity are referred to as RS mutant recombinant purified antibodies. In one embodiment, these RS mutant recombinant purified antibodies further comprise a mutation for binding to Protein L.

[0117] The antibodies of the present disclosure can have mutations in their constituent amino acid sequences, such as their constant region, heavy chain variable region, light chain variable region, heavy chain CDR1-3, or light chain CDR1-3, to the extent that their antigen-binding properties and physiological activity are not lost. Such mutations may be, but are not limited to, substitutions, deletions, insertions, and the like. For example, conservative substitution techniques can be employed for substitutions. Furthermore, by analyzing the binding mode between the antibody and the antigen in detail using three-dimensional structural analysis or the like, various mutations can be introduced into the antibodies of the present disclosure to the extent that their antigen-binding properties are not lost.

[0118] In one embodiment, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure may be provided in the form of an antibody consisting of one heavy chain and one light chain (also called a single-chain antibody), in addition to a form consisting of two heavy chains and two light chains. Furthermore, the antibody according to the present disclosure may be provided in the form of a dimer or multimer having, in addition to two heavy chains and two light chains, a J (joining) chain and, optionally, an SC (secretary component).

[0119] When the antibodies of the present disclosure are used as multimers, mutants or fusion proteins with mutations beneficial to multimerization may be used. For example, when using the w27 antibody as a multimer, the mutant described in WO 2024 / 038839 in which cartilage oligomeric matrix protein (COMP) is fused to the C-terminus of the heavy chain can be used. In one embodiment, a multimer-forming IgA having the configuration described in WO 2024 / 038839 as the "notail-comp" configuration, i.e., the W27 antibody and CHO cell-produced recombinant IgA of the W27 antibody described in WO 2014 / 142084, which lack the tail piece of the H chain of RS_H000_L001 (W27 IgA) in which a mutation for protein L binding has been introduced into the light chain, can be used, in which the COMP sequence is fused to the C-terminus of the H chain without a linker.

[0120] In one embodiment, the antibody or antigen-binding fragment thereof used in the combination of the present disclosure includes one in which the structure of its constant or variable region has been altered from that of the native antibody, to the extent that its antigen-binding properties and biological activity are not lost. In one embodiment, such alterations include amino acid substitution, deletion, insertion, etc. (addition to the terminal). In one embodiment, such alterations include fusion or conjugation with other peptide or non-peptide structures. In one embodiment, the structure of the constant or variable region thus altered has at least 80%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity in its amino acid portion with the native amino acid sequence.

[0121] (Nucleic Acid and Nucleic Acid Set) In one aspect, the present disclosure provides a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present disclosure that does not bind to microorganisms beneficial to the recipient organism and has the effect of improving the intestinal microbiota. The nucleic acid encoding the antibody or antigen-binding fragment thereof may be ribonucleotides or deoxynucleotides. Furthermore, the form of the nucleic acid is not particularly limited, and may be single-stranded or double-stranded. The codons used in the nucleic acid sequence are not particularly limited, and various codons can be appropriately selected and used depending on the purpose. For example, appropriate codons can be selected during production taking into consideration codon frequency and the type of recipient cell to be administered. In one aspect, a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present disclosure is used to express and produce an antibody or antigen-binding fragment thereof of the present disclosure. In one aspect, a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present disclosure is used to express an antibody or antigen-binding fragment thereof of the present disclosure in vivo. In one aspect, an antibody or antigen-binding fragment thereof of the present disclosure is administered as a nucleic acid encoding it and a nucleic acid set.

[0122] When a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present disclosure encodes an antigen-binding fragment, for example, the two domains of the Fv fragment, VL and VH, may be encoded by separate nucleic acid molecules, or, using recombinant techniques, a single protein chain in which the pair of VL and VH domains form a monovalent molecule (single-chain Fv (scFv)) may be encoded by a single nucleic acid.

[0123] In one embodiment, a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present disclosure may be in the form of a vector. For example, the vector may be an expression vector or a vector for use in genetic recombination. The configuration of the vector is not particularly limited and may include various components used in known vectors.

[0124] In one embodiment, a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present disclosure encodes a portion of the peptide fragment that constitutes the antibody or antigen-binding fragment, and the nucleic acid may be provided as a set together with nucleic acids encoding other peptide fragments that constitute the antibody or antigen-binding fragment. The set of nucleic acids may include all of the nucleic acids encoding the peptide fragments that constitute the antibody or antigen-binding fragment, or may include only a portion of the nucleic acids encoding the peptide fragments that constitute the antibody or antigen-binding fragment. A set of nucleic acids containing only a portion of the nucleic acids can be used in combination with another nucleic acid or set of nucleic acids that encodes another portion of the peptide fragment that constitutes the antibody or antigen-binding fragment.

[0125] (Form and Use of Composition) The form and method of use of the composition used in the combination of the present disclosure are not particularly limited. The composition of the present disclosure can be used as a medicine, as well as in the fields of food and feed. Therefore, the composition of the present disclosure can be a pharmaceutical composition, a food composition, or a feed composition. The food composition includes not only general foods, but also foods for specified health uses, nutritional supplements, functional foods, foods for the sick, etc.

[0126] In one aspect, when the composition of the present disclosure is provided as a pharmaceutical composition, the composition may be provided for use in improving the intestinal microbiota of a subject organism, imparting or enhancing an immunostimulatory effect in a subject organism, an anti-inflammatory effect in a subject organism, or a microbicidal effect in a subject organism, or for preventing or treating a symptom or disease requiring the imparting or enhancing of these effects.

[0127] In one aspect, imparting or enhancing the intestinal microbiota-improving effect of the subject organism includes increasing the diversity of the microbiota in the gastrointestinal contents or excrement of the subject organism. Also, in another aspect, the intestinal microbiota-improving effect of the subject organism includes increasing the amount of short-chain fatty acids in the gastrointestinal contents or excrement of the subject organism.

[0128] Imparting or enhancing an immunostimulatory effect in a target organism includes enhancing immune function in the target organism. In one embodiment, the immunostimulatory effect in a target organism includes enhancing intestinal immune function in the target organism.

[0129] Imparting or enhancing an anti-inflammatory effect in a subject organism includes suppressing an inflammatory response in the subject organism. In one embodiment, the anti-inflammatory effect in a subject organism includes suppressing an inflammatory response in the intestinal tract of the subject organism.

[0130] Imparting or enhancing a microbicidal effect in a target organism includes killing or inhibiting the growth of microorganisms that adversely affect the target cells, or promoting the excretion of such microorganisms from the body.

[0131] Symptoms or diseases that require improvement of the gastrointestinal bacterial flora of a target organism, or the imparting and promoting of immunostimulatory, anti-inflammatory, and bactericidal effects in the target organism, include, for example, intestinal microbial symbiotic imbalance (dysbiosis), inflammatory bowel disease (IBD), and bacterial enteritis.

[0132] In one aspect, the compositions of the present disclosure are suitable for use in preventing or treating inflammatory bowel disease (IBD) and bacterial enteritis in subjects with a gut dysbiosis imbalance.

[0133] The method for determining the presence or absence of an intestinal microbial dysbiosis imbalance is not particularly limited, and methods commonly known in the art can be used. For example, a method for determining the presence or absence of an intestinal microbial dysbiosis imbalance can be used by determining 16S RNA or other nucleic acid sequences of bacteria in the subject's stool or intestinal mucosa (Appl Environ Microbiol. 2021 May 11;87(11):e00395-21. doi: 10.1128 / AEM.00395-21).

[0134] In one aspect, when the composition of the present disclosure is provided as a food composition or feed composition, the composition may be provided with labeling relating to the improvement of the intestinal microflora of a subject organism, the imparting or enhancing of an immunostimulatory effect in a subject organism, an anti-inflammatory effect in a subject organism, or a microbicidal effect in a subject organism, or the prevention or treatment of a symptom or disease requiring the imparting or enhancing of these effects.

[0135] When the composition of the present disclosure contains an antibody or an antigen-binding fragment thereof, or a microorganism beneficial to the organism to which it is administered, it may contain an effective amount of these.

[0136] As used herein, the term "effective amount" refers to an amount that, when administered in combination with an antibody or antigen-binding fragment of the present disclosure or a microorganism beneficial to a target organism, is effective in improving the intestinal microflora of the target organism, imparting or enhancing an immunostimulatory effect in the target organism, an anti-inflammatory effect in the target organism, or a microbicidal effect in the target organism, or in preventing or treating a symptom or disease requiring the imparting or enhancement of these effects.

[0137] The dose of the antibody or antigen-binding fragment thereof, or the microorganism beneficial to the recipient organism, contained in the composition of the present disclosure can be appropriately determined taking into consideration the condition of the recipient, the degree of effect exerted by administration, the dosage form, the administration method, the recipient, the gender of the recipient, etc.

[0138] When an antibody of the present disclosure is included, the dosage of the antibody can be appropriately set so that it typically ranges from 0.001 to 100 mg / kg / day on average. In some embodiments, the dose is the amount administered over a specific period (e.g., daily or weekly). By administering a combination of a microorganism and an antibody, the composition of the present disclosure can achieve the desired effect even at an extremely low dose of the antibody. Therefore, the amount of an antibody or antigen-binding fragment thereof contained in a composition of the present disclosure can be 1 / 2 to 1 / 20, 1 / 3 to 1 / 20, 1 / 4 to 1 / 20, 1 / 5 to 1 / 20, 1 / 6 to 1 / 20, 1 / 7 to 1 / 20, 1 / 8 to 1 / 20, 1 / 10 to 1 / 20, 1 / 15 to 1 / 20, 1 / 2 to 1 / 15, 1 / 2 to 1 / 10, 1 / 2 to 1 / 9, 1 / 2 to 1 / 8, 1 / 2 to 1 / 7, 1 / 2 to 1 / 6, 1 / 2 to 1 / 5, 1 / 2 to 1 / 4, 1 / 2 to 1 / 3, 1 / 3 to 1 / 15, 1 / 4 to 1 / 10, 1 / 5 to 1 / 8, etc., of the amount when the antibody is used alone.

[0139] When the compositions of the present disclosure include microorganisms beneficial to the subject, the amount of microorganism in the composition can be expressed by weight, number of microorganisms, and / or CFU (colony forming units). In some aspects, the compositions disclosed herein contain each microorganism of the composition in an amount of about 10, about 10 per dose. ２ , about 10 ３ , about 10 ４ , about 10 ５ , about 10 ６ , about 10 ７ , about 10 ８ , about 10 ９ , about 10 １０ , about 10 １１ , about 1012, about 10 １３ In some aspects, the compositions disclosed herein contain about 10, about 10 total microorganisms per dose. ２ , about 10 ３ , about 10 ４ , about 10 ５ , about 10 ６ , about 10 ７ , about 10 ８ , about 10 ９ , about 10 １０ , about 10 １１ , about 10 １２ , about 10１３ Contains more than one.

[0140] The composition of the present disclosure may contain two or more different microorganisms as beneficial microorganisms to the subject of administration. When the composition contains two or more different microorganisms, they may be present in different amounts in the composition. For example, as a non-limiting example, the composition may contain 10 microorganisms A and 10 microorganisms B and C. ３ 10 microorganisms B ４ and microorganism C to 10 ６ In some aspects, the compositions disclosed herein may contain about 10, about 10, or about 10 of each microorganism in the composition per dose. ２ , about 10 ３ , about 10 ４ , about 10 ５ , about 10 ６ , about 10 ７ , about 10 ８ , about 10 ９ , about 10 １０ , about 10 １１ , about 10 １２ , about 10 １３ In some aspects, the compositions disclosed herein contain more than about 10 CFU of the total microorganisms combined per dose. １ , about 10 ２ , about 10 ３ , about 10 ４ , about 10 ５ , about 10 ６ , about 10 ７ , about 10 ８ , about 10 ９ , about 10 １０ , about 10 １１ , about 10 １２ , about 10 １３ As noted above, the microorganisms of the composition may be present in different amounts. In some aspects, the compositions disclosed herein contain each microorganism of the composition at about 10 CFU per dose. －７ , about 10 －６ , about 10 －５ , about 10 －４ , about 10 －３ , about 10 －２ , about 10 －１ In some aspects, the compositions disclosed herein contain more than about 10 g of total combined microorganisms per dose.－７ , about 10 －６ , about 10 －５ , about 10 －４ , about 10 －３ , about 10 －２ , about 10 －１ In some embodiments, a dose is the amount in one administration device (e.g., one tablet, pill, or capsule). In some embodiments, a dose is the amount administered over a specific period of time (e.g., one day or one week).

[0141] The amount of the antibody or antigen-binding fragment of the present disclosure, or the microorganism beneficial to the target organism, contained in the composition of the present disclosure can be appropriately determined taking into consideration the administration form, administration dose, etc.

[0142] The compositions of the present disclosure may contain pharmaceutically acceptable carriers or additives in combination with the antibodies or antigen-binding fragments, or microorganisms beneficial to the subject of administration. For example, in the case of pharmaceutical compositions, the pharmaceutically acceptable carriers or additives refer to any carrier, diluent, excipient, suspending agent, lubricant, adjuvant, vehicle, delivery system, emulsifier, tablet disintegrant, absorbent, preservative, surfactant, colorant, flavoring, or sweetener, and any known pharmaceutically acceptable carrier or additive may be used.

[0143] In one embodiment, the composition of the present disclosure may contain ingredients that are beneficial to the growth, survival, and health maintenance of microorganisms that are beneficial to the subject of administration. In one embodiment, the composition of the present disclosure may contain food ingredients, such as indigestible oligosaccharides and dietary fiber, that are generally called prebiotics, which reach the large intestine without being broken down or absorbed in the stomach or small intestine and serve as food for microorganisms that live in the large intestine.

[0144] The organisms to which the composition of the present disclosure is administered are not particularly limited, and examples include mammals such as humans, primates, mice, rats, guinea pigs, rabbits, hamsters, dogs, cats, weasels, cows, and pigs, and birds such as chickens.

[0145] The dosage and administration method of the composition of the present disclosure can be appropriately determined taking into consideration the condition of the subject, the degree of effect exerted by administration, the dosage form, the administration method, the subject, the gender of the subject, etc.

[0146] The administration method is not particularly limited, but direct administration into the digestive tract is preferred, and examples of such administration methods include oral administration, nasal administration, transmucosal administration, and enteral administration.

[0147] Enteral administration is not limited to administration via the anus, but also includes administration via a tube or the like inserted into the digestive tract from outside the individual, such as a gastrostomy. The location into which the digestive tract is inserted is not limited to the intestine, but includes the esophagus, stomach, small intestine (including the duodenum, jejunum, ileum, etc.), large intestine (including the cecum, colon, rectum, etc.), etc.

[0148] The composition according to the present disclosure may be administered in the above amount once a day or in divided doses. Furthermore, the administration interval may be daily, every other day, weekly, biweekly, every 2-3 weeks, monthly, bimonthly, or every 2-3 months, as long as it has a preventive or therapeutic effect against the above diseases.

[0149] When a microorganism beneficial to a target organism, or an antibody or antigen-binding fragment thereof having an intestinal microbiota-improving effect that does not bind to the microorganism beneficial to the target organism, used in combination with a composition of the present disclosure, is contained in a composition different from the composition of the present disclosure, the administration schedule, such as the order, interval, and frequency of administration of these compositions, is not particularly limited. For example, a composition containing the antibody and a composition containing microorganisms beneficial to the target organism may be administered simultaneously. In another aspect, the composition containing the antibody may be administered once daily, and the composition containing microorganisms beneficial to the target organism may be administered three times daily.

[0150] When the composition of the present disclosure is provided as a food composition, its specific form is not particularly limited.For example, the composition of the present disclosure can be provided as a food itself or as a food additive to be incorporated into a food composition.More specifically, the composition can be provided in the form of beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, milk drinks, etc.; frozen desserts such as ice cream, ice sorbet, shaved ice, etc.; confectioneries such as candy, candy, gum, chocolate, tablet candy, snacks, biscuits, jelly, jam, cream, baked goods, etc.; noodles such as soba, udon, harusame, Chinese noodles, instant noodles, etc.; processed seafood and livestock foods such as kamaboko, ham, sausage, etc.; dairy products such as processed milk and fermented milk, oils and fats and oil-based foods such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing, etc.; seasonings such as sauces, sauces, etc.; soups, stews, salads, side dishes, sprinkles, pickles, bread, cereals, etc. In the case of foods for specified health uses, nutritional supplements, functional foods, etc., they may be provided in the form of powder, granules, capsules, lozenges, tablets, syrup, etc.

[0151] When the composition of the present disclosure is provided as a feed composition, its specific form is not particularly limited. For example, as long as the effects of the feed composition according to the present disclosure are not impaired, the feed composition may be prepared by mixing the composition with a normal feed or, if necessary, mixing it with components that can be incorporated into a normal feed, or the feed composition itself may be used as feed.

[0152] (Method for Producing Composition) In one aspect, the present disclosure provides a method for producing the composition of the present disclosure. In one aspect, the present disclosure provides the following method for producing the composition of the present disclosure. The method for producing a composition comprising the antibody or antigen-binding fragment thereof, and to be administered in combination with microorganisms beneficial to the target organism, comprises the steps of: preparing microorganisms that have been administered, are being administered, or are intended to be administered to the target organism; obtaining an antibody or antigen-binding fragment thereof that does not bind to the microorganism and has the effect of improving the intestinal microbiota; and formulating the antibody or antigen-binding fragment thereof with an excipient that can be used in pharmaceutical, food, or feed compositions.

[0153] In one aspect, the present disclosure provides a composition comprising an antibody having an intestinal flora-improving effect, produced by the above-described production method.

[0154] In one aspect, the present disclosure provides the following method for producing a composition of the present disclosure: A method for producing a composition comprising microorganisms beneficial to a target organism and to be administered in combination with an antibody or antigen-binding fragment thereof having an intestinal flora-improving effect, the composition comprising: a step of preparing an antibody or antigen-binding fragment thereof having an intestinal flora-improving effect that has been administered, is being administered, or is intended to be administered to a target organism; a step of preparing microorganisms that do not bind to the antibody and are beneficial to the target organism; and a step of formulating the microorganisms beneficial to the target organism together with an excipient that can be used in pharmaceutical, food, or feed compositions.

[0155] In one aspect, the present disclosure provides a composition containing microorganisms beneficial to a target organism, produced by the above-described production method.

[0156] 2. Method of Using in Combination Microorganisms Beneficial to a Target Organism and an Antibody Having an Intestinal Microbiota-Improvement Activity That Does Not Bind to the Microorganisms Beneficial to the Target Organism In one aspect, the present disclosure provides a method of using in combination microorganisms beneficial to a target organism and an antibody or an antigen-binding fragment thereof that has an intestinal microbiota-improving activity that does not bind to the microorganisms beneficial to the target organism.

[0157] In one aspect, the present disclosure provides the following method: A method for treating a subject, comprising administering to the subject a microorganism beneficial to the subject, and an antibody or an antigen-binding fragment thereof having an effect of improving the intestinal microbiota, which does not bind to the microorganism beneficial to the subject.

[0158] In one aspect, the method further comprises the step of confirming, before administering the antibody or antigen-binding fragment thereof, that the antibody or antigen-binding fragment thereof does not bind to microorganisms beneficial to the organism to which the antibody or antigen-binding fragment is to be administered.

[0159] In one aspect, the present disclosure provides the following method: A method for treating a subject, comprising the step of administering to the subject an antibody or antigen-binding fragment thereof having an activity of improving the intestinal microbiota, the antibody or antigen-binding fragment ...

[0160] In one aspect, the method further comprises the step of confirming, before administering the antibody or antigen-binding fragment thereof, that the antibody or antigen-binding fragment thereof does not bind to microorganisms beneficial to the organism to which the antibody or antigen-binding fragment is to be administered.

[0161] In one embodiment, the method further comprises administering beneficial microorganisms to the recipient organism.

[0162] In one aspect, the present disclosure provides the following method: A method for treating a subject who has been administered, is currently being administered, or is scheduled to be administered an antibody or antigen-binding fragment thereof having an activity of improving the intestinal microbiota, comprising the step of administering beneficial microorganisms to the subject.

[0163] In one aspect, the method further comprises a step of confirming, before administering beneficial microorganisms to the target organism, that the beneficial microorganisms do not bind to the antibody or antigen-binding fragment thereof having the effect of improving the intestinal microbiota.

[0164] In one aspect, the above method further comprises the step of administering to the recipient organism an antibody or antigen-binding fragment thereof that has an intestinal flora-improving effect.

[0165] (Microorganisms beneficial to the target organism) The microorganisms beneficial to the target organism used in the method of combining an antibody of the present disclosure with a microorganism beneficial to the target organism are not particularly limited, and include microorganisms that have a beneficial effect on the survival, health, etc. of the target organism. More specifically, the microorganisms beneficial to the target organism used in the method of the present disclosure can be those described above in "1. Combination composition of a microorganism beneficial to the target organism and an antibody that does not bind to the microorganism beneficial to the target organism and has an intestinal bacterial flora-improving effect" (Microorganisms beneficial to the target organism).

[0166] (Antibody having an effect of improving intestinal flora) The antibody or antigen-binding fragment thereof used in the method of the present disclosure is not particularly limited, as long as it is an antibody or antigen-binding fragment thereof that has an effect of improving intestinal flora and does not bind to microorganisms that are beneficial to the target organism.

[0167] More specifically, the antibody or antigen-binding fragment thereof used in the method of the present disclosure can be the antibody or antigen-binding fragment thereof described in the above section "1. Combination composition of a microorganism beneficial to the target organism and an antibody having an intestinal flora-improving effect that does not bind to the microorganism beneficial to the target organism" (Antibody having an intestinal flora-improving effect).

[0168] (Microorganisms that have an adverse effect on the target organism) In one aspect, the antibody or antigen-binding fragment thereof used in the method of the present disclosure binds to microorganisms that have an adverse effect on the target organism, as explained in the above section (Microorganisms that have an adverse effect on the target organism) of "1. Combination composition of microorganisms that are beneficial to the target organism and antibodies that have an intestinal flora-improving effect and do not bind to the microorganisms that are beneficial to the target organism."

[0169] In one embodiment, the antibody or antigen-binding fragment thereof used in the methods of the present disclosure is administered as a nucleic acid encoding the same and a set of nucleic acids.

[0170] (Target of administration)

[0171] The target organisms in the methods of the present disclosure are not particularly limited, but include, for example, mammals such as humans, primates, mice, rats, guinea pigs, rabbits, hamsters, dogs, cats, weasels, cows, and pigs, and birds such as chickens.

[0172] (Mode of Administration) In the method of the present disclosure, antibodies or microorganisms beneficial to the target organism can be administered as various compositions. More specifically, these compositions can be those described in (Form and Use of Composition) of "1. Combination Composition of Microorganism Beneficial to the Target Organism and Antibody That Binds to Microorganisms That Do Not Bind to the Microorganism Beneficial to the Target Organism, but Have a Negative Effect on the Target Organism." The dosage and method of administration of the composition of the present disclosure can be appropriately determined taking into consideration the condition of the target organism, the degree of effect exerted by administration, the dosage form, the method of administration, the target organism, the gender of the target, etc.

[0173] The administration method in the method of the present disclosure is not particularly limited, but direct administration to the digestive tract is preferred, and examples of such administration methods include oral administration, nasal administration, transmucosal administration, and enteral administration.

[0174] Enteral administration is not limited to administration via the anus, but also includes administration via a tube or the like inserted into the digestive tract from outside the individual, such as a gastrostomy. The location into which the digestive tract is inserted is not limited to the intestine, but includes the esophagus, stomach, small intestine (including the duodenum, jejunum, ileum, etc.), large intestine (including the cecum, colon, rectum, etc.), etc.

[0175] The administration in the method of the present disclosure may be once a day or divided into several doses. Furthermore, the administration interval may be daily, every other day, weekly, biweekly, every 2-3 weeks, monthly, bimonthly, or every 2-3 months, as long as it has a preventive or therapeutic effect against the above-mentioned disease.

[0176] In one embodiment, the method of the present disclosure may further administer to the subject a component beneficial to the growth, survival, and health maintenance of microorganisms beneficial to the subject. In one embodiment, the method of the present disclosure includes administering to the subject food components, such as indigestible oligosaccharides and dietary fiber, generally referred to as prebiotics, which reach the large intestine without being broken down or absorbed in the stomach or small intestine and serve as food for microorganisms living in the large intestine.

[0177] In the methods of the present disclosure, the administration schedule, such as the order, interval, and frequency of administration of an antibody or antigen-binding fragment thereof that binds to a microorganism beneficial to the target organism or a microorganism that does not bind to a microorganism beneficial to the target organism but has a deleterious effect on the target organism, is not particularly limited. For example, the antibody or antigen-binding fragment thereof and the microorganism beneficial to the target organism may be administered simultaneously. In another aspect, the antibody or antigen-binding fragment thereof may be administered once daily, and the microorganism beneficial to the target organism may be administered three times daily.

[0178] In one aspect, the methods of the present disclosure are used to prevent or treat a condition or disease in a subject having such a condition or disease requiring the imparting or enhancement of a gut microbiota-improving, immunostimulatory, anti-inflammatory, or microbicidal effect, or the imparting or enhancement of such an effect.

[0179] In one aspect, imparting or enhancing the intestinal microbiota-improving effect of the subject organism in the method of the present disclosure comprises increasing the bacterial flora diversity in the gastrointestinal contents or excrement of the subject organism. Also, in another aspect, the intestinal microbiota-improving effect of the subject organism comprises increasing the amount of short-chain fatty acids in the gastrointestinal contents or excrement of the subject organism.

[0180] In one aspect, imparting or enhancing an immunostimulatory effect to a target organism in the method of the present disclosure comprises enhancing immune function in the target organism. In one aspect, the immunostimulatory effect in the target organism comprises enhancing intestinal immune function in the target organism.

[0181] In one aspect, imparting or enhancing an anti-inflammatory effect to a subject organism in the method of the present disclosure comprises suppressing an inflammatory response in the subject organism. In one aspect, the anti-inflammatory effect in a subject organism comprises suppressing an inflammatory response in the intestinal tract of the subject organism.

[0182] In one aspect, imparting or enhancing the microbicidal effect of the target organism in the method of the present disclosure includes killing or inhibiting the growth of microorganisms that adversely affect the target cells, or promoting the excretion of such microorganisms from the body.

[0183] In one aspect, symptoms or diseases requiring improvement of the gastrointestinal flora of a subject organism to which the method of the present disclosure is applied, imparting and promoting immunostimulatory, anti-inflammatory, or bactericidal effects in the subject organism, or imparting or promoting these effects, include, for example, inflammatory bowel disease (IBD) and bacterial enteritis.

[0184] In one aspect, the methods of the present disclosure are suitable for use in preventing or treating intestinal dysbiosis, inflammatory bowel disease (IBD), or bacterial enteritis.

[0185] In one aspect, the methods of the present disclosure are suitable for use in preventing or treating inflammatory bowel disease (IBD) in a subject with a gut dysbiosis imbalance, or bacterial enteritis in a subject with a gut dysbiosis imbalance.

[0186] In one aspect, the method of the present disclosure includes a step of confirming that the subject has or is at risk of a gut microbial dysbiosis imbalance before administering the antibody or antigen-binding fragment thereof, or the microorganism.

[0187] In one aspect, when the composition of the present disclosure is provided as a food composition or feed composition, the composition may be provided with labeling relating to the improvement of the intestinal microflora of a subject organism, the imparting or enhancing of an immunostimulatory effect in a subject organism, an anti-inflammatory effect in a subject organism, or a microbicidal effect in a subject organism, or the prevention or treatment of a symptom or disease requiring the imparting or enhancing of these effects.

[0188]

[0189] Hereinafter, the present disclosure will be described in more detail with reference to examples, but these are merely illustrative and do not limit the present disclosure.

[0190] Example 1: Antibody that binds to microorganisms that have a detrimental effect on the target organism but not to microorganisms that are beneficial to the target organism

[0191] (Materials and Methods) Echerichia coli, Clostridioides difficile, Clostridium butyricum, Clostridium saccharolyticum, Clostridium sindens, and Bifidobacterium breve were each cultured anaerobically at 37°C (Brain Heart Infusion media, 80% N ２ , 10% H ２ , 10% CO ２ ) and Lactobacillus paracasei was cultured at 37°C aerobically (MRS medium). After the culture, the cells were collected by centrifugation. The collected bacteria were then resuspended in 0.05M Na ２ CO ３ The antibody was suspended in buffer and coated onto an ELISA plate. After blocking with 1% BSA-added PBS, the IgA antibody RS_H000_L001 (rW27) with intestinal flora-improving activity or a control IgA antibody (Control IgA) was added and reacted at room temperature for approximately 1 hour. The plate was then washed with 0.05% Tween 20-added PBS, and a secondary detection antibody was added and reacted. After the reaction, a color reaction was carried out using an Alkali Phosphatase tablet (Sigma). To ensure sufficient reaction, the ELISA plate after color development was reacted overnight at 4°C, and then incubated with TriStar. ２ LB942 (BERTHOLD TECHNOLOGIES) is used, and ４０５ nm was measured.

[0192] (Results) The antibody RS_H000_L001 (rW27), which has the effect of improving the intestinal flora, bound to Echerichia coli and Clostridioides difficile, which have adverse effects on the target organism, and also to Clostridium butyricum and Clostridium saccharolyticum. On the other hand, it was confirmed that it did not bind to the beneficial bacteria Clostridium sindens, Lactobacillus paracasei, and Bifidobacterium breve. By administering the antibody RS_H000_L001 (rW27) having an intestinal flora-improving effect in combination with microorganisms beneficial to the target organism, such as Clostridium sindens, Lactobacillus paracasei, and Bifidobacterium breve, it is possible to improve the gastrointestinal microbiota of the target organism, and to impart and promote immunostimulatory, anti-inflammatory, and bactericidal effects to the target organism. Thus, it is understood that by administering an antibody having an intestinal flora-improving effect in combination with microorganisms beneficial to the target organism to which the antibody does not bind, it is possible to obtain synergistically excellent effects in the prevention or treatment of symptoms or diseases requiring the improvement of the gastrointestinal microbiota of the target organism, the impartation and promotion of immunostimulatory, anti-inflammatory, and bactericidal effects to the target organism, or the impartation or promotion of these effects.

[0193] Example 2: C. difficile mouse infection experiment

[0194] (Materials and Methods) (1) Preparation of C. difficile Spore Suspension A spore suspension of the C. difficile strain (VPI10483) was prepared, aliquoted, and stored at -80°C. The spore suspension was thawed and cultured, and then inoculated onto a C. difficile selective medium plate (TCCFA plate), and the number of viable bacteria obtained was determined in advance.

[0195] C. difficile was plated on SMC medium and cultured anaerobically at 37°C for 7 days. 1 ml of PBS was added to the plate, and colonies were scraped with a cell scraper and transferred to a 1.5 ml tube. After centrifugation at 13,200 rpm and 4°C for 2 minutes, the supernatant was discarded and the colonies were suspended in 1 ml of ice-cold sterile water. This procedure was repeated five times, followed by suspension in 1 ml of ice-cold sterile water and storage at -20°C for at least 48 hours. After thawing, the colonies were centrifuged at 13,200 rpm and 4°C for 2 minutes, the supernatant was discarded, and the colonies were suspended in 1 ml of ice-cold sterile water. This procedure was repeated three times. After another 2 minutes of centrifugation at 13,200 rpm and 4°C, the supernatant was discarded and the colonies were suspended in 1 ml of 1% BSA / PBS solution. This procedure was repeated three times. After heating at 65°C for 30 minutes, 100 µl of the mixture was dispensed into 30 1.5 mL tubes and stored at -80°C. The composition of the SMC medium is shown below.

[0196] The above reagents were dissolved in 200 ml of sterile distilled water and sterilized in an autoclave. After cooling to about 60°C, 600 μl of 10% (w / v) L-cysteine ​​was added, and the mixture was dispensed into 10 cm plates.

[0197] C. difficile selective medium plates were prepared by dissolving the reagents listed in Table 1 below in 400 ml of sterile distilled water and sterilizing the solution in an autoclave. The solution was allowed to cool to 60°C, and then 2 ml of Cycloserine (Sigma-Aldrich) (50 mg / ml) and 2 ml of Cefoxitin (Sigma-Aldrich) (1.6 mg / ml) were added. Appropriate amounts were then dispensed onto 10 cm plates.

[0198] (Table 1) Composition of C. difficile selective medium

[0199] (2) Preparation of Lactic Acid Bacteria Synprotec (a food containing live lactic acid bacteria and bifidobacteria) sold by Yakult Co., Ltd. was added to MRS medium and MRC medium and cultured overnight at 37 ° C under anaerobic conditions. Since bacterial growth was confirmed in each medium, the culture solution was plated on MRS agar medium and MRC agar medium and cultured again under anaerobic conditions at 37 ° C for two nights. The 16S rDNA sequences of the colonies formed on the agar medium were analyzed, and the colonies on the MRS medium were identified as Lacticaseibacillus paracasei, and the colonies on the MRC medium were identified as Bifidobacterium breve. The isolated lactic acid bacteria and bifidobacteria were prepared as glycerol stocks and stored at -80 ° C. When each bacterium was inoculated onto MRS medium from a glycerol stock, it was confirmed that both bacteria could grow. Therefore, it was decided to use MRS medium, which is easy to prepare, for the administration experiment. The compositions of the MRS and MRC media used for the culture are as follows. Composition of MRS medium (Difco Lactobacilli MRS Broth, BD#288130)

[0200] The MRS medium used was Difco Lactobacilli MRS Broth (#288130) sold by BD. Following the instructions on the product, 55 g of this product was dissolved in 1,000 ml of sterile distilled water and sterilized in an autoclave. MRC medium composition The above reagents were dissolved in 1,000 ml of sterile distilled water and sterilized in an autoclave. When preparing the agar medium, agar powder was added to each medium at a ratio of 15 g / l. After autoclave sterilization, the mixture was cooled to about 60°C, dispensed into sterilized petri dishes, and allowed to solidify.

[0201] Preparation of IgA The immunoglobulin used was a polymeric IgA having the [notail-comp] structure described in WO / 2024 / 038839, i.e., the W27 antibody described in WO2014 / 142084 and a CHO cell-produced recombinant IgA of the W27 antibody, RS_H000_L001 (W27 IgA), in which a mutation for protein L binding was introduced into the light chain and the tail piece of the H chain was deleted, with the COMP sequence fused to the C-terminus of the H chain without a linker.

[0202] The amino acid sequence of the H chain of this exemplary antibody and the nucleotide sequence encoding it are shown below: SEQ ID NO: 114: CHO W27notailCOMP IgA H amino acid sequence (without signal sequence) >SEQ ID NO: 115: Nucleic acid sequence encoding CHO W27notailCOMP IgA H (including N-terminal signal sequence)

[0203] The light chain of this IgA antibody has the same amino acid sequence as the light chain of a modified CHO cell-produced recombinant IgA of the W27 antibody (RS_H000_L001) (see WO 2023 / 277142), and the amino acid sequence of the light chain variable region (RS_LV001) has a mutation for Protein L binding. The amino acid sequence of this light chain is shown below. >SEQ ID NO: 116: RS_LV001

[0204] The gene encoding the IgA and the vector containing it were prepared and introduced into CHO cells according to standard methods, similar to the method described in Example 8 of WO2023 / 277166.

[0205] (4) Administration to mice C57BL / 6 mice (8 weeks old) were purchased from CLEA Japan and acclimated for one week in a sterile isolator for infection experiments. After a week of acclimation, a triple antibiotic mixture (gentamicin: 500 mg / kg, kanamycin: 150 mg / kg, metronidazole: 50 mg / kg) was orally administered to the mice using a probe for four days. After four days, the weight of each mouse was measured, and mice that lost weight due to antibiotic administration were excluded. The remaining mice were randomly divided into groups. The day after the antibiotic administration was completed, C. difficile spores (1x10 ３ Each mouse was orally infected with 1.0 x 10^7 cells of L. paracasei and B. breve (cfu) using a probe. Six hours later, 50 μg of the test antibody was orally administered using a probe. Subsequently, 50 μg of antibody was administered once daily for a total of seven days. From day 8 onwards, the mice were observed without antibody administration. Six hours after C. difficile spore administration, the lactic acid bacteria-administered group was orally administered a mixture of 1.0 x 10^7 cells of L. paracasei and B. breve together with PBS or 50 g of the test antibody. The day of C. difficile administration was counted as day 0, and the mice were administered once daily for a total of seven days until day 6. From day 7 onwards, administration was discontinued and the mice were observed. During this period, the mice were weighed, feces observed, and feces collected. After infection, feces collected at each time point were suspended in PBS in an amount corresponding to the weight of the feces. An equal volume of ethanol was added to the suspension, and the suspension was vortexed and allowed to stand at room temperature for 1 hour. Further serial dilutions were then performed with PBS. These diluted bacterial solutions were plated on C. difficile selective medium plates and cultured anaerobically to determine the number of viable bacteria per gram of feces.

[0206] (5) Measurement of C. difficile Toxin B Clostridium difficile Toxin A OR B quanti (tgcBioMICS # TGC-E002-1) was used. The collected feces were suspended in an amount of PBS according to the weight, and the suspension was used as a stock solution or diluted 10 times. The reaction and measurement were carried out according to the kit's instructions, and the amount of Toxin B in 1 mL of fecal suspension was determined.

[0207] (6) Extraction of Bacterial DNA DNA lysis buffer for DNA extraction was prepared by adding Tris-HCl (pH 8.0, final concentration 50 mM), NaCl (final concentration 300 mM), EDTA (final concentration 1 mM), and 0.5% SDS to ultrapure water. The collected bacterial cells were suspended in 500 μl of DNA lysis buffer. After suspension, the cells were transferred to a 2.0 ml tube (TM-625S, TOMY) containing glass beads (GB-01, 0.1Φ, TOMY) and subjected to bead disruption at 3,500 rpm for 1 minute using a bead cell disrupter (MS-100, TOMY). Proteinase K (final concentration 0.5 mg / ml, Nacalai Tesque) was added to the bead-disrupted bacterial solution, vortexed, and allowed to stand overnight at 55°C to completely lyse the bacterial cells. The temperature was returned to room temperature from 55°C, and 500 μl of phenol-chloroform-isoamyl alcohol (Nacalai Tesque) was added to the bacterial cell lysate, mixed by vortexing, and centrifuged at 13,000 rpm for 5 minutes at 4°C. The upper layer was collected in another 1.5 ml tube. An equal volume of chloroform (Nacalai Tesque) was added, mixed by vortexing, and centrifuged at 13,000 rpm for 5 minutes at 4°C. The upper layer was again collected in another 1.5 ml tube, and equal volumes of isopropanol (Nacalai Tesque) and glycogen (final concentration 0.04 mg / ml, Nacalai Tesque) were added and mixed by inversion. The DNA solution was mixed by inversion and allowed to stand at -20°C for 20 minutes, then centrifuged at 13,000 rpm for 10 minutes at 4°C, and the supernatant was removed. 500 μl of 70% ethanol was added to the precipitated DNA, and the mixture was centrifuged at 13,000 rpm for 5 minutes at 4°C, and the supernatant was removed. The precipitated DNA was air-dried, and then Ultra Pure Distilled Water (Invitrogen) was added to dissolve the DNA.

[0208] (7) 16S ribosomal RNA (rRNA) gene amplicon sequence analysis of bacterial DNA. Bacterial DNA was subjected to polymerase chain reaction (PCR) targeting the V3-V4 region of the variable region of the 16S rRNA gene. Primers were prepared for next-generation sequencing using Miseq, and index sequences were added to distinguish the samples to be analyzed. The composition of the PCR reaction solution is shown in Table 1, and the PCR reaction is shown in Table 2. After the PCR reaction, 10x Loading Buffer (TAKARA) was added, and agarose electrophoresis was performed. The gel used to run the PCR samples was a 1.5% agarose gel (VWR Life Science) supplemented with LED Stain G (LABTAS+). A 100 bp DNA Ladder (TAKARA) was used as a DNA size marker. Electrophoresis was performed at 100 V for 30 minutes. After electrophoresis, approximately 500 bp PCR products were excised and purified using a Fast Gene Gel Extraction Kit (Nihon Genetics). A portion of the purified PCR product was electrophoresed again to confirm whether it was purified. Sequence analysis was performed by Myskin Inc. using Miseq Reagent Kit V3 (Illumina). The sequence data obtained by sequence analysis was subjected to bacterial flora analysis (phylogenetic analysis, alpha diversity analysis, beta diversity analysis) using Qiime2 software (ver. 2020-2). SILVA and Greengenes databases were used. For α diversity analysis, the Shannon index was calculated, and for β diversity analysis, principal coordinate analysis (PCoA) based on Jaccard and unweighted uniFrac distances was performed. Table 1 Table 2

[0209] (Results) When lactic acid bacteria and IgA antibodies were administered alone in a C. difficile-infected mouse model, weight loss was observed to be comparable to that observed with PBS. The group administered lactic acid bacteria alone experienced slightly faster weight recovery than PBS, but the IgA-only group also experienced a similar weight recovery rate (Figure 4). On the other hand, the group administered lactic acid bacteria and IgA antibodies in combination significantly suppressed weight loss, and the slight weight loss observed on Day 5 showed a tendency toward recovery by the following day (Figure 4). This slight weight loss indicates that inflammatory responses caused by C. difficile infection were suppressed from the onset stage. When the survival rates of mice administered these test drugs were examined, administration of 0.05 mg / mouse of IgA antibodies alone did not improve survival rates at all, but all mice in the group administered lactic acid bacteria and IgA antibodies in combination survived (Figure 5). In a similar model mouse study, it was confirmed that an IgA antibody similar to that used in this experiment improved the survival rate to 100% when administered at a dose of 0.3 mg once a day (WO2024 / 185747).

[0210] Next, the fecal C. difficile counts of mice administered these test drugs were examined. On Day 9, no significant differences in fecal C. difficile counts were observed between the test groups. On Day 14, however, a significant decrease in fecal C. difficile counts was observed in the group administered lactic acid bacteria in combination with IgA antibodies (Figure 6, upper panel). The fecal Toxin B concentrations of mice administered these test drugs were examined. On Day 11, a significant decrease in fecal Toxin B concentration was observed in the group administered lactic acid bacteria in combination with IgA antibodies (Figure 6, lower panel).

[0211] When the intestinal flora of mice administered these test drugs was examined, the intestinal flora in the group administered lactic acid bacteria alone on Day 4 showed a significantly different intestinal flora composition than the other test groups (Figure 7) and a decrease in diversity (Figure 8). This result is thought to be due to the special effects of the interaction between lactic acid bacteria and the intestinal bacterial symbiotic imbalance (dysbiosis) caused by antibiotic administration before C. difficile infection. As mentioned above, although there was no decrease in survival rate in the group administered lactic acid bacteria alone, it is thought that the interaction between dysbiosis and lactic acid bacteria caused weight loss and diarrhea.

[0212] The results of this example demonstrate that the combined administration of lactic acid bacteria and IgA antibodies provides a qualitatively different and superior infection prevention effect compared to the administration of either agent alone. The combined administration of lactic acid bacteria and IgA antibodies was shown to suppress diarrheal symptoms caused by the administration of bacteria alone in a dysbiotic state. Furthermore, this combined administration acts at an earlier stage, rapidly eliminating toxins from infectious bacteria and highly effectively suppressing symptoms such as diarrhea. It is understood that the combined administration of microorganisms beneficial to the target organism and IgA antibodies can achieve excellent infection prevention effects by using extremely low doses of IgA antibodies. While lower doses of antibody drugs such as IgA are desirable from the perspective of suppressing side effects, this combined administration has excellent properties as a pharmaceutical.

[0213] The above examples demonstrate that the combination of a microorganism beneficial to a target organism and an antibody having an intestinal microbiota-improving effect, as disclosed herein, exhibits excellent effects in the prevention or treatment of symptoms or diseases requiring the impartation or enhancement of an intestinal microbiota-improving effect in a target organism, an immunostimulatory effect in a target organism, an anti-inflammatory effect in a target organism, or a microbicidal effect in a target organism, or the impartation or enhancement of these effects, particularly in the prevention or treatment of intestinal microbial symbiotic imbalance (dysbiosis), inflammatory bowel disease (IBD), or bacterial enteritis.

Claims

1. A composition comprising an antibody or its antigen-binding fragment that has the effect of improving the intestinal flora and does not bind to microorganisms that are beneficial to the target organism, characterized in that it is administered in combination with microorganisms that are beneficial to the target organism.

2. A composition comprising microorganisms beneficial to the target organism, which is administered in combination with an antibody or its antigen-binding fragment that has the effect of improving the intestinal flora and does not bind to the microorganisms beneficial to the target organism.

3. The composition according to claim 1 or 2, wherein the microorganisms beneficial to the target organism are selected from the group consisting of microorganisms that produce short-chain fatty acids, microorganisms that suppress the production of secondary bile acids, microorganisms that suppress the growth of microorganisms harmful to the target organism, and microorganisms that suppress the production of toxins from microorganisms that have a negative effect on the target organism.

4. The composition according to claim 1 or 2, wherein the microorganisms beneficial to the organism to which the composition is administered are Clostridium sindens, Lactobacillus paracasei, and / or Bifidobacterium breve.

5. The composition according to claim 1 or 2, wherein the antibody binds to a microorganism that adversely affects the subject to which it is administered.

6. The composition according to claim 1 or 2, wherein the microorganisms that adversely affect the organism to which the composition is administered are microorganisms associated with diseases or microorganisms that produce toxins.

7. The composition according to claim 1 or 2, wherein the microorganism that adversely affects the organism to which the composition is administered is Escherichia coli or Clostridioides difficile.

8. The antibody is: a) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:8; b) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:17, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:18; c) a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:27, and an antibody comprising a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO:28; d) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO:81, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO:82; e) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO:93, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO:94;f) an antibody comprising a heavy chain variable region comprising the amino acid sequence of heavy chain CDR1, the amino acid sequence of heavy chain CDR2, and the amino acid sequence of heavy chain CDR3 of a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 37, and a light chain variable region comprising the amino acid sequence of light chain CDR1, the amino acid sequence of light chain CDR2, and the amino acid sequence of light chain CDR3 of a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 38; or g) an antibody which has at least one amino acid mutation in at least one region selected from heavy chain CDR1 to 3, light chain CDR1 to 3, and light chain FR1 relative to a reference antibody comprising a heavy chain variable region comprising the amino acid sequence represented by SEQ ID NO: 37 and a light chain variable region comprising the amino acid sequence represented by SEQ ID NO: 38, and which binds to the amino acid sequence RQEEHIELIAS in Escherichia coli SHMT protein and the amino acid sequence VLDMMMKLEKPE in C. difficile iPGM protein.

9. The composition according to claim 1 or 2, which is a composition for use in the prevention or treatment of symptoms or diseases requiring the imparting or enhancement of an intestinal microflora improving effect, an immunostimulatory effect in a subject to be administered, an anti-inflammatory effect in a subject to be administered, or a microbicidal effect in a subject to be administered.

10. The composition according to claim 1 or 2, which is a composition for use in the prevention or treatment of intestinal microbial dysbiosis, inflammatory bowel disease (IBD) or bacterial enteritis.

11. The composition according to claim 1 or 2, which is for use in the prevention or treatment of inflammatory bowel disease (IBD) or bacterial enteritis associated with an imbalance in intestinal microbial dysbiosis.

12. The composition of claim 1 or 2, further administered in combination with an oligosaccharide.

13. The composition according to claim 1 or 2, wherein the composition contains a microorganism beneficial to the target organism and the antibody or antigen-binding fragment thereof in the same composition.

14. The composition according to claim 1 or 2, comprising a microorganism beneficial to the organism to which it is administered and the antibody or antigen-binding fragment thereof in separate compositions.

15. The composition of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is contained in the composition as a nucleic acid or set of nucleic acids encoding the antibody or antigen-binding fragment thereof.

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