Antitoxin antibodies and their use

Abstract

The present disclosure provides antibodies and antigen-binding fragments that can be administered to subjects bitten by venomous snakes.The antibodies or antigen-binding fragments herein can be capable of treating or curing subjects, and can provide protection from snake venom for up to several weeks.When the type of snake is unknown, or when a subject is bitten by more than one species of snake, a combination or population of antibodies and antigen-binding fragments can be administered to the subject.The present disclosure further provides a method for identifying such broadly neutralizing antibodies.

Description

Technical Field

[0001] Cross-reference This application claims the benefit of U.S. Provisional Application No. 63 / 172,782, filed Apr. 9, 2021, which is hereby incorporated by reference in its entirety.

[0002] Sequence Listing This application includes a sequence listing submitted electronically in ASCII format, which is hereby incorporated by reference in its entirety. The ASCII copy created in 2022 is named 60333-704601_SL.txt and is of size __ bytes.

[0003] Description of Research Funded by the Federal Government This invention was made with government support under Grant No. 1R43AI147898-01 awarded by the National Institutes of Health. The U.S. government has certain rights in this invention.

Summary of the Invention

Means for Solving the Problems

[0004] Abstract This application discloses universal, broad-spectrum neutralizing fully human antitoxin antibodies and cocktails of antibodies that exhibit properties far superior to those currently available. References to antibodies herein can be interchanged with antigen-binding fragments. Antitoxin antibodies and cocktails of antibodies that can neutralize toxins from multiple snake species and that have a reduced potential for negative side effects are disclosed. Also disclosed are populations of antibodies containing multiple copies of a single antibody that can broadly neutralize snake venoms from more than one species of snake or can broadly neutralize snake venoms from more than one genus of snake. The present disclosure further provides methods for the identification of such broad-spectrum neutralizing antibodies.

[0005] Such broad-spectrum neutralizing monoclonal antibodies effective against all classes of toxins across multiple snake species enable the prospect of a single universal antitoxin against all snake species. Broad-spectrum neutralizing antibodies (bnAbs) are necessary for antibody-based universal antitoxins because there are over 550 venomous snake species, and each toxin consists of 10 to 70 specific toxic proteins. Therefore, without antitoxin bnAbs, generating 5,500 to 38,500 specific antibodies specialized for each toxin from every species, let alone combining them so that they exist individually at concentrations sufficient to provide any meaningful protection, would be an impractically ambitious task. However, with antitoxin bnAbs, it is possible to have individual antibodies that neutralize the shared common variant toxin homologs found across all snake venoms from all species. Snake venom toxins belong to 10 distinct toxin classes, and only the "Big Four" (long-chain neurotoxins, PLA2, SVSP, and SVMP) of these cause illness and death in humans from snake bites. Therefore, in principle, a universal antitoxin could consist of a cocktail of just four broad-spectrum neutralizing antitoxin monoclonal antibodies.

[0006] Our research has recovered Centi-D9, a broadly neutralizing, ultra-high affinity antibody against long-chain neurotoxins, which was able to neutralize long-chain neurotoxins from cobra, taipan, and mamba and provide in vivo protection therefrom. We obtained the crystal structure of Centi-D9 in complex with long-chain neurotoxins from cobra, taipan, krait, and mamba, and this crystal structure revealed the common broadly neutralizing mechanism of these evolutionarily related, but distinct, toxin family members. Most importantly, we demonstrated that Centi-D9 as a monoclonal antibody can independently provide complete in vivo protection of mice from whole toxin challenges for black mamba and several cobra species, as well as partial protection against taipan. The results of these studies demonstrate the possibility of universal antitoxin.

[0007] In 2021, snakebite-induced venom injections continued to rank on the World Health Organization's list of neglected tropical diseases, claiming between 81,000 and 138,000 lives annually due to their hematotoxic or neurotoxic effects, and resulting in up to 300,000 amputations or permanent disabilities each year (2018 reported figures). Snakebite injections affect every continent except Antarctica, with the highest incidences in Asia, Africa, and South America, disproportionately impacting rural populations (i.e., agricultural workers) and children. Snakebite injections also affect the United States, with 7,000 to 8,000 cases annually, including an average of 50 cases from imported non-native snake species. 59.3% of these injections result in moderate to severe outcomes and an average of 3 deaths. It's a global problem, but major pharmaceutical companies haven't invested in developing treatments for poison injections, and in recent years have even abandoned the market due to the wide range of species that would fragment it.

[0008] Unlike existing approaches, the strategy proposed by the inventors involves isolating and characterizing a broadly neutralizing, fully human antitoxin antibody that is stable, field-use, and does not require refrigeration. Due to the low immunogenicity of fully human IgG, this approach significantly reduces the potential for adverse effects such as anaphylactic shock, which are common with existing therapies using animal-derived antitoxins. Importantly, this therapy targets multiple snake species geographically, thus eliminating the need for snakebite victims to identify the species that injected the venom. These molecules can be optimized to have a better safety profile and potentially higher efficacy. They are also designed to be heat-stable, thereby enabling field use for rapid treatment during critical minutes after a snakebite. Due to less stringent storage requirements and a longer shelf life, this therapy makes the treatment available at a more competitive price point, thereby greatly expanding the types of medical centers that can store and utilize it, and enabling access to a wider and more diverse population.

[0009] The technology described herein utilizes a novel and diverse immunotherapy library collected from a middle-aged man who has undergone dose-escalating autoimmunization (over 700 boosts and 200 raw bites) over the past 20 years from some of the world's most venomous snakes, including the mamba, taipan, krait, viper, rattlesnake, and cobra. Toxins from multiple snake species were panned against our library over several rounds to identify cross-reactive clones. Preliminary studies revealed 282 unique clones enriched with VH CDR3, indicating potential cross-reactive binding factors in these output pools, across different toxins. This is highly significant, as cross-reactive antibodies are essential for the development of broad-spectrum neutralizing antitoxins. With the discovery of Centi-D9, which targets long-chain alpha-neurotoxins, we have already demonstrated the success of developing a single therapeutic or prophylactic antibody capable of protecting against all toxins in multiple types of elapid snakes (e.g., taipan, mamba, cobra).

[0010] The development of broad-spectrum vaccines has been limited by both the many different species and the toxin diversity in the toxins of each species. For over a century, treatment for venom injection from snake bites has typically involved horse or sheep serum therapy using animal-derived antitoxin preparations containing either immunoglobulin G (IgG) or derivative antigen-binding fragments (Fab) from a single toxin. More recently, several sheep or horse-derived serum products with a somewhat broader range of species applicability have been developed (CroFab for American pit vipers, EchiTAb for West African sawtooth vipers or carpet vipers). While effective in reducing mortality and permanent disability in many cases, heterologous antitoxin serum therapy presents several challenges. Depending on the antitoxin, 6–59% of patients experience early adverse reactions to non-human horse or sheep plasma-derived antitoxins, including early adverse anaphylactic reactions (within 24 hours). Antitoxins consist of approximately 70% Fab or Ig against antigens encountered by immunized animals, rather than the toxic components. This reduces their potency and necessitates more extensive drug infusion, typically via intravenous (IV) administration in a hospital setting, which may not be close to the site of poisoning. This results in significant transportation problems, leading to the patient dying before reaching the hospital or being unable to be treated by the time they arrive.

[0011] Polyclonal Fab-based formulations can lead to treatment with shortened half-lives and inconsistent batch quality compared to monoclonal IgG. Digestion into Fab fragments reduces serum sickness by removing non-human Fc regions, but these treatments shorten half-lives from weeks to hours, thus requiring 8–10 doses of antitoxin. However, patients will have a severe reaction initiated to these antibodies and will not be able to receive any additional future doses. High concentrations of IgG can cause adverse reactions, including inflammation and serum sickness, 1–2 weeks after treatment. Finally, antitoxins developed for a single species require precise identification of the specific snake that bit the victim, which is extremely difficult for victims and medical personnel unfamiliar with the snake phenotype, and necessitate clinics stockpiling the exact toxins of many possible products for different venom injections.

[0012] The antitoxin compositions described herein are for broad-spectrum defense against major families of snakes with lethal venom. In addition, the treatment does not require definitive identification of the venom-injecting species, can be administered rapidly after a snake bite, or prophylactically to individuals at high risk of snake bites. This will reduce costs due to the elimination of the need to stockpile multiple antitoxins and the extended half-life of the lyophilized formulation of Centivax. Snake venom injection is traditionally a niche market resembling a rare disease, a highly unmet need with under-invested resources, particularly by large pharmaceutical companies. However, the crucial difference is that snake venom injection is not rare, with 5 million injections, 300,000 permanent injuries, and 125,000 deaths annually. Since the market is currently fragmented across multiple venoms of snake species, a single agent has the potential to consolidate and expand the existing market for antitoxins. The primary end-users and customers will be the armed forces of the top 25 countries, where deployed military personnel can utilize the inventors' antitoxin as an effective preventative or acute treatment option in harsh environments. Further uses include stockpiling to many of the 28,600 hospitals worldwide, and with government subsidies to support distribution in severely affected countries.

[0013] Non-human immunotherapy. For over a century, treatment for venom injection from snake bites has typically involved horse or sheep serum therapy using animal-derived antitoxin preparations containing derivative antigen-binding fragments (Fabs) from a single toxin. More recently, several sheep or horse-derived serum products with a somewhat broader range of species applicability have been developed. CroFab (sheep-derived antitoxin, polyvalent and lyophilized) and AnaVip (horse-derived Fab2, lyophilized powder for injection) are both specific to North American snakes: rattlesnakes, swamp vipers, and American pit vipers. EchiTAb-G is a sheep antiserum effective only against West African saw-toothed snakes or carpet vipers in sub-Saharan Africa, and is recommended for use in sub-Saharan Africa. While effective in reducing mortality and permanent disability in many cases, heterologous antitoxin serum therapy presents several challenges. Depending on the antitoxin, 6–59% of patients experience early adverse reactions due to non-autologous reactions to exogenous equine or sheep plasma-derived antitoxins. These adverse reactions may include early adverse anaphylactic reactions (within 24 hours).

[0014] Traditional antitoxins require IV injection. These antitoxins consist of approximately 70% Fab or Ig against other antigens rather than the toxic component. This has several drawbacks, as it reduces their potency and requires larger drug infusions. The drugs are often delivered IV at a site other than the site of toxic injection, thus leading to increased mortality due to the time delay in administration. IV drugs are severely limited in their use for treatment due to the time delay, the required healthcare personnel, and the site of treatment. Polyclonal Fab-based formulations can lead to treatment with shortened half-lives and inconsistent batch quality compared to monoclonal IgG. Digestion into Fab fragments reduces serum sickness by removing the non-human Fc region, but these treatments shorten the half-life from weeks to hours, thus requiring 8-10 doses of antitoxin. Furthermore, additional administration of these antibodies cannot be performed due to the predicted serological toxicity. High concentrations of non-human IgG antitoxins can cause inflammation and serum sickness 1-2 weeks after treatment.

[0015] To date, antitoxins composed of monoclonal IgG have not been used in clinical practice. While other antitoxin companies, such as Venomyx, have demonstrated some cross-reactivity in preliminary studies, they still derive their molecules from immunized camels, thus requiring costly and time-consuming humanization. Furthermore, their single-domain nature may result in a shortened half-life compared to fully human IgG. If not adequately neutralized with a single dose, repeated injections are necessary due to their small size, which allows for renal excretion.

[0016] On the other hand, the technology described herein utilizes a novel and diverse immunization library taken from a middle-aged man who has undergone dose-escalating autoimmunization (over 700 boosts and 200 raw bites) over the past 17 years from some of the world's most venomous snakes, including the mamba, taipan, krait, viper, rattlesnake, and cobra.

[0017] Library construction utilized a unique next-generation sequencing (NGS) technique to amplify antibody variable domains from blood samples taken before and 7 days after final toxicology immunization in male subjects, followed by deep sequencing and antibody phage display to allow tracing downstream toxin-binding factors back to the source blood samples. Antibody candidates against toxins from black mamba, western diamondback rattlesnake, and coastal taipan were isolated during the initial library panning.

[0018] Heat-stabilized antibodies. Standard antitoxins require continuous refrigeration at 2–8°C to maintain stability. This poses a significant challenge to arid villages, often inhabited by locally endemic venomous species, which may lack the infrastructure to properly store antitoxins. Recent research has investigated the stability of antitoxins at room temperature. This application will develop heat-stabilized antibodies with less stringent storage requirements and longer shelf lives, which are expected to be available at a more competitive price point and thus greatly expand the types of medical centers that can store and utilize them. Antitoxin cross-reactivity is a complex biochemical challenge, despite the fact that all medically relevant snakes belong to three families: Atractaspidae, Viperidae, and Elapidae, with the vast majority belonging to either Viperidae or Elapidae. Within these groups, the toxin proteome of their toxins is classified into 8–11 subfamilies, four of which are the major components across all toxins. Some existing antitoxins function well in certain snake lineages despite 8 million years of isolation, while other, more closely related species exhibit poor neutralization to the same antitoxins because a single amino acid mutation in the toxin molecule is sufficient to interfere with antibody recognition.

[0019] Preliminary deep sequencing panning analysis of the inventors' human subject antibody under the inventors' control demonstrated multi-species VH CDR3 domain enrichment, indicating potential cross-reactivity candidates to multiple snake venom species. It is reasonable to assume that the inventors' subject has produced an antibody capable of binding to an epitope conserved across relevant peptides between snake species, given that it has been immunized with various snakes from the Viperidae and Elapidae families for nearly 20 years. This application isolates and characterizes a broadly neutralizing, fully human antitoxin antibody using high-throughput surface plasmon resonance (SPR), full-spectrum fluorescence and light scattering, as well as in vivo functional assays, to develop a broad-spectrum antitoxin.

[0020] Broad-spectrum neutralizing antibodies (bnAbs). There are 550+ venomous snake species, and each toxin consists of 10 to 70 specific toxic proteins. To develop a single universal antitoxin, broad-spectrum neutralizing monoclonal antibodies would need to be effective against all classes of toxins across multiple snake species. Without antitoxin bnAbs, generating 5,500 to 38,500 specific antibodies specialized for each toxin from every species, let alone combining them so that they exist individually at concentrations sufficient to provide any meaningful protection, would be an impractically ambitious task. However, with antitoxin bnAbs, it is possible to have individual antibodies that neutralize the shared common variant toxin homologs found across all snake venoms from all species. Snake venom toxins belong to 10 distinct toxin classes, and only the "Big Four" of these (long-chain neurotoxins, PLA2, SVSP, and SVMP) cause disease and death in humans from snake bites; therefore, a universal antitoxin can consist of a cocktail of only four broad-spectrum neutralizing antitoxin monoclonal antibodies (Figure 11).

[0021] Broad-spectrum neutralizing antibodies will break the limitations of antitoxin therapy. 1) Broad-spectrum neutralizing antibodies can recognize the entire family of snake toxins, thus enabling a single antitoxin product for each lethal snake family. This is dramatically beneficial in snake identification and avoiding the risk of mis-antitoxin assessment, ultimately strengthening the market economy by producing a single product for regions worldwide. 2) Human monoclonal antibodies are not rejected as immunogens by the recipient's immune system, so fully human antibodies can avoid serotoxin toxicity by reducing the occurrence of adverse reactions. These molecules could be easily optimized to have a better safety profile and potentially higher efficacy. Monoclonal IgGs have a favorable effect against snake toxins in terms of their neutralizing ability against several specific toxins involved in myonecrosis and proteolytic effects in in vivo lethality studies, neutralization tests, and other applications. Recombinant antitoxins can be 100% pure antitoxin-specific products and therefore can be much more potent, thereby reducing volume and allowing delivery by two-chamber syringes outside of a hospital setting. Due to its improved toxicity profile compared to sheep IgG, full human IgG can be offered as a prophylactic agent with a 3-6 week protective period, making it suitable for individuals in areas and / or occupations with a high risk of exposure. Recombinant antibodies and antibody fragments of human origin are attractive as alternatives to antitoxin production.

[0022] Fully Human Antitoxin Antibodies. This application describes the development of safer and more effective next-generation antitoxin biotherapeutics by isolating and characterizing a broad-spectrum, fully human antitoxin antibody cocktail for treating venom injections resulting from snake bites. Monoclonal fully human-derived IgG significantly reduces the potential for adverse effects commonly seen with animal-derived antitoxins and allows for greater optimization, which is expected to be more cost-effective than conventionally produced antitoxins.

[0023] Universal antitoxin compositions comprising a population of antitoxin antibodies or antigen-binding fragments are provided herein. The population of antitoxin antibodies or antigen-binding fragments may comprise one or one or more antibodies or antigen-binding fragments. Alternatively, the population of antitoxin antibodies or antigen-binding fragments may comprise two antibodies or antigen-binding fragments, three antibodies or antigen-binding fragments, four antibodies or antigen-binding fragments, five antibodies or antigen-binding fragments, six antibodies or antigen-binding fragments, or more antibodies or antigen-binding fragments.

[0024] One or more antibodies or antigen-binding fragments may contain, be derived from, IgG, IgM, IgE, IgA, or IgD, or a combination thereof. One or more antibodies or antigen-binding fragments may contain monoclonal antibodies, graft antibodies, chimeric antibodies, human antibodies, or humanized antibodies. Antigen-binding fragments may contain Fab, Fab', F(ab')2, variable fragment (Fv), triabody, tetrabody, minibody, bispecific F(ab')2, triplicate F(ab')2, diabody, bispecific diabody, single-chain variable fragment (scFv), scFv-Fc, Fab-Fc, VHH, or bispecific scFv.

[0025] One or more antibodies or antigen-binding fragments include VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3. One or more antibodies or antigen-binding fragments may include VH CDR1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 2. Alternatively, or in addition, one or more antibodies or antigen-binding fragments may include VH CDR2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 3. Alternatively, or in addition, one or more antibodies or antigen-binding fragments include a VH CDR3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 1. Alternatively, or in addition, one or more antibodies or antigen-binding fragments include a VL CDR1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 4. Alternatively, or in addition, one or more antibody or antigen-binding fragments include VL CDR2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 5. Alternatively, or in addition, one or more antibody or antigen-binding fragments include VL CDR3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 6.

[0026] Alternatively, or in addition, one or more antibodies or antigen-binding fragments include FW-H1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 7. Alternatively, or in addition, one or more antibodies or antigen-binding fragments include FW-H2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 8. Alternatively, or in addition, one or more antibody or antigen-binding fragments include FW-H3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 9. Alternatively, or in addition, one or more antibody or antigen-binding fragments include FW-H4 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 10.

[0027] Alternatively, or in addition, one or more antibody or antigen-binding fragments include FW-L1 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 11. Alternatively, or in addition, one or more antibody or antigen-binding fragments include FW-L2 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 12. Alternatively, or in addition, one or more antibody or antigen-binding fragments include FW-L3 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 13. Alternatively, or in addition, one or more antibody or antigen-binding fragments include FW-L4 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 14.

[0028] Alternatively, or in addition, one or more antibody or antigen-binding fragments include VH having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 15. Alternatively, or in addition, one or more antibody or antigen-binding fragments include VL having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 16.

[0029] In another embodiment, a method for treating a subject suffering from poison injection is provided herein, comprising the step of administering to the subject a composition comprising an effective amount of a universal antitoxin comprising one or more antibodies or antigen-binding fragments. In some cases, poison injection is caused by one or more species of snakes.

[0030] The method may include the step of administering one or more antibodies or antigen-binding fragments to a subject. Alternatively, the method may include the step of administering to the subject one antibody, two antibodies or antigen-binding fragments, three antibodies or antigen-binding fragments, four antibodies or antigen-binding fragments, five antibodies or antigen-binding fragments, six antibodies or antigen-binding fragments, seven antibodies or antigen-binding fragments, eight antibodies or antigen-binding fragments, nine antibodies or antigen-binding fragments, ten antibodies or antigen-binding fragments, or more antibodies or antigen-binding fragments. The one or more antibodies or antigen-binding fragments to be administered to the subject may include, or are derived from, IgG, IgM, IgE, IgA, or IgD, or a combination thereof. The one or more antibodies or antigen-binding fragments to be administered to the subject may include monoclonal antibodies, graft antibodies, chimeric antibodies, human antibodies, or humanized antibodies. The antigen-binding fragments administered to the target may include Fab, Fab', F(ab')2, variable fragment (Fv), triabody, tetrabody, minibody, bispecific F(ab')2, tripspecific F(ab')2, diabody, bispecific diabody, single-chain variable fragment (scFv), scFv-Fc, Fab-Fc, VHH, bispecific scFv, or combinations thereof.

[0031] In any of these methods, the subject may be administered one or more additional therapies or drugs. These additional therapies or drugs may include, for example, NSAIDs. Alternatively, or in addition, the additional therapies or drugs may include, for example, PLA2 inhibitors. PLA2 inhibitors include, but are not limited to, valesprazib, methylvalesprazib, or combinations thereof. Administration may be obtained by any preferred means, such as one or more injections.

[0032] In any of such methods, one or more antibodies or antigen-binding fragments to be administered to a subject may include a VH CDR3 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 1. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to a subject may include a VH CDR1 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 2. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject contain a VH CDR2 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 3.

[0033] Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include VL CDR1 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 4. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include VL CDR2 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 5. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject contain a VL CDR3 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 6.

[0034] Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-H1 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 7. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-H2 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 8. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-H3 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 9. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-H4 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 10.

[0035] Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-L1 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 11. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-L2 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 12. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-L3 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 13. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include FW-L4 having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 14.

[0036] Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include VH having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 15. Alternatively, or in addition, one or more antibodies or antigen-binding fragments to be administered to the subject include VL having an amino acid sequence that is at least approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 16.

[0037] In any such example of a universal antitoxin composition or method, one or more antibodies or antigen-binding fragments are found in Boiga irregularis (Southern Giant Snake), Boiga cyanea (Green Giant Snake), Boiga dendrophila (Mangrove Snake), Dispholidus typus (Boomslang Snake), Salvadora grahamiae (Mountain Patchnose Snake), Spalerosophis diadema (Crowned Snake), Tantilla nigriceps (Plains Blackhead Snake), Thelotornis capensis (Southern Twig Snake), Thelotornis kirtlandii (Northern Twig Snake), Toxicodryas blandingii (Branding Tree Snake), Trimorphodon lambda (Sonoran Lyre Snake), Amphiesma stolatum (Buffalo Kaleback), Natrix Tessellate (dice snake), Rhabdophis subminiatus (red-necked keelback), Rhabdophis tigrinus (Japanese keelback), Thamnophis elegans (Western terrestrial garter snake), Thamnophis sirtalis (common garter snake), Ahaetulla nasuta (long-nosed whip snake), Atractaspis bibronii (bibronze's burrowing asp), Atractaspis dahomeyensis (Dahomey's burrowing asp), Atractaspis engaddensis (Palestinian mole snake), Atractaspis microlepidota (small-scaled burrowing asp), Malpolon monspessulanus (Montpellier's snake), Acanthophis antarcticus (common death adder), Aipysurus laevis (olive brown sea snake), Aipysurus duboisii (Dubois sea snake), Austrelaps superbus (lowland pit viper), Cryptophis nigrescens (small ice snake), Demansiaolivacea (Olive Whip Snake), Emydocephalus annulatus (Spotted Sea Snake), Furina tristis (Stevens' Striped Snake), Hydrophis melanocephalus (Black-headed Sea Snake), Hydrophis curtus (Spiny Sea Snake), Hydrophis gracilis (Slender Sea Snake), Hydrophis elegans (Elegant Sea Snake), Hydrophis jerdonii (Corn-noseed Sea Snake), Hydrophis klossi (Slangor Sea Snake), Hydrophis peronii (Horned Sea Snake), Hydrophis belcheri (Belcher's Sea Snake), Hydrophis stricticollis (Bengal Sea Snake), Hydrophis major (Olive-headed Sea Snake), Hydrophis stokesii (large-headed sea snake), Hydrophis melanosoma (black-banded robust sea snake), Hydrophis hardwickii (dorsal-ventral sea snake), Hydrophis cyanocinctus (spotted sea snake), Hydrophis spiralis (narrow-banded sea snake), Hydrophis nigrocinctus (black-banded sea snake), Hydrophis platurus (black-backed sea snake), Hydrophis ornatus (black-spotted sea snake), Hydrophis viperinus (venomous sea snake), Hydrophis schistosus (wart sea snake), Notechis scutatus (mainland tiger snake), Oxyuranus scutellatus (coastal taipan), Oxyuranus temporalis (central range taipan), Pseudechis australis (Margas snake), Pseudechis butleri (Butler's black snake), Pseudechis colletti (Colette's black snake), Pseudechis guttatus (Blue-bellied black snake), Pseudechis papuanus (Papua black snake), PseudechisPorphyriacus (Red-bellied Black Snake), Pseudonaja affinis (Dugait's Snake), Pseudonaja guttata (Speckled Brown Snake), Pseudonaja inframacula (Peninsula Brown Snake), Pseudonaja nuchalis (Western Brown Snake), Pseudonaja textilis (Eastern Brown Snake), Tropidechis carinatus (Clarence River Snake), Aspidelaps lubricus (Cape Coral Snake), Aspidelaps scutatus (Shield-nose Snake), Bungarus fasciatus (Round-tailed Krait), Bungarus caeruleus (Indoor Krait), Bungarus candidus (Blue Krait), Bungarus flaviceps (Red-headed Crate), Bungarus multicinctus (Krait), Dendroaspis viridis (Western Green Mamba), Dendroaspis angusticeps (Eastern Green Mamba), Dendroaspis jamesoni (Jameson's Mamba), Dendroaspis polylepis (Black Mamba), Elapsoidea sundevallii (Sandebaru African Garter Snake), Hemachatus haemachatus (Lincalus), Laticauda colubrina (Blue-spotted Sea Snake), Laticauda laticaudata (Broad-skinned Sea Snake), Laticauda semifasciata (Erabu Sea Snake), Micrurus obscurus (Bolivian Coral Snake), Micrurus frontalis (Southern Coral Snake), Micrurus alleni (Allen's Coral Snake), Micrurus altirostris (Uruguayan Coral Snake), Micrurus clarki (Clark's Coral Snake), Micrurus corallinus (painted coral snake), Micrurus distans (Western Mexican coral snake), Micrurus dumerilii (Dumeril's coral snake), MicrurusMicrurus fulvius (Eastern coral snake), Micrurus hemprichii (Hemprich's coral snake), Micrurus ibiboboca (Kaachinga coral snake), Micrurus lemniscatus (South American coral snake), Micrurus mipartitus (Red-tailed coral snake), Micrurus mosquitensis (Costa Rican coral snake), Micrurus multifasciatus (Striped coral snake), Micrurus nigrocinctus (Central American coral snake), Micrurus pyrrhocryptus (Argentine coral snake), Micrurus spixii (Amazonian coral snake), Micrurus surinamensis (Aquatic coral snake), Micrurus tener (Texas coral snake), Micrurus tschudii (Desert coral snake), Naja siamensis (Indochinese spitting cobra), Naja Naja annulata (Ringed Water Cobra), Naja annulifera (Long-nosed Cobra), Naja ashei (Giant Poisonous Cobra), Naja atra (Taiwanese Cobra), Naja christyi (Conglomerate Water Cobra), Naja haje (Egyptian Cobra), Naja kaouthia (Thai Cobra), Naja katiensis (Western African Poisonous Cobra), Naja melanoleuca (Forest Cobra), Naja mossambica (Mozambican Poisonous Cobra), Naja (Indian Cobra), Naja nigricollis (Black-necked Poisonous Cobra), Naja nivea (Cape Cobra), Naja nubiae (Nubian Poisonous Cobra), Naja oxiana (Caspian Cobra), Naja pallida (Red Poisonous Cobra), Naja philippinensis (Northern Philippine Cobra), Naja Samarensis (Samara cobra), Naja sputatrix (Indonesian spitting cobra), Naja sumatrana (Sumatran spitting cobra), Ophiophagus hannah (King cobra), Walterinnesia aegyptia (Western desert cobra), Homalopsisbuccata (Linnaeus's water snake), Myrrophis chinensis (Chinese mud snake), Subsessor bocourti (Bocourt's water snake), Azemiops feae (cobra viper), Agkistrodon bilineatus (patterned pit viper), Agkistrodon contortrix (American pit viper), Agkistrodon piscivorus (swamp pit viper), Agkistrodon taylori (castellana), Agkistrodon laticinctus (broadband copperhead), Atropoides picadoi (Picado's jumping pit viper), Bothriechis lateralis (sidestriped palm pit viper), Bothriechis nigroviridis (black spotted palm pit viper), Bothriechis schlegelii (eyelash palm pit viper), Bothrops diporus (chacolance head), Bothrops erythromelas (Curtin Lancehead), Bothrops insularis (Golden Lancehead Viper), Bothrops jararaca (Jararaka), Bothrops neuwiedi (Neuwiedi Lancehead), Bothrops pauloensis (Blackfaced Lancehead), Bothrops asper (Terciopero), Bothrops atrox (Kaikasa), Bothrops ayerbei (Ayerbei Lancehead), Bothrops caribbaeus (Saint Lucia Lancehead), Bothrops jararacussu (Jararacus), Bothrops lanceolatus (Martiny Lancehead), Bothrops leucurus (Whitetail Lancehead), Bothrops moojeni (Brazil Lancehead), Bothrops alternatus (Urutu), Bothrops cotiara (Cotiara), Bothrops fonsecai (Lance Head of Fonseca), Bothrops itapetiningae (Lance Head of São Paulo), Bothropstaeniatus (Speckled Forest Pit Viper), Bothrops mattogrossensis (Matogrosso Lanzen Otter), Calloselasma rhodostoma (Malayan Pit Viper), Cerrophidion godmani (Godman Mountains Pit Viper), Cerrophidion sasai (Costa Rican Mountains Pit Viper), Crotalus viridis (Prairie Rattlesnake) Crotalus atrox (Western diamondback rattlesnake), Crotalus adamanteus (Eastern diamondback rattlesnake), Crotalus basiliscus (Mexican west coast rattlesnake), Crotalus catalinensis (Santa Catalina Island rattlesnake), Crotalus cerastes (Leftback rattlesnake), Crotalus Cerberus (Arizona black rattlesnake), Crotalus durissus (South American rattlesnake), Crotalus enyo (Baja California rattlesnake), Crotalus horridus (Wood rattlesnake), Crotalus lepidus (Mottled rock rattlesnake), Crotalus mitchellii (San Lucan spotted rattlesnake), Crotalus molossus (Northern black-tailed rattlesnake), Crotalus oreganus (North Pacific rattlesnake), Crotalus Pricei (Western Twin-Spotted Rattlesnake), Crotalus pusillus (Tacita Land Rattlesnake), Crotalus ravus (Mexican Pygmy Rattlesnake), Crotalus ruber (Red Diamond Rattlesnake), Crotalus scutulatus (Mojave Rattlesnake), Crotalus simus (Central American Rattlesnake), Crotalus tigris (Tiger Rattlesnake), Crotalus totonacus (Totonacan Rattlesnake), Crotalus tzabcan (Yucatan Neotropical Rattlesnake), Crotalus willardi (Arizona Ridgenose Rattlesnake), Crotalus Pyrrhus (Southwestern Speckled Rattlesnake), Crotalus vegrandis (Uracoan Rattlesnake), Deinagkistrodon acutus (Hyappoda), Gloydius intermedius (intermediate pit viper), Gloydius blomhoffii (pit viper), Gloydius brevicaudus (long-tailed pit viper), Gloydius halys (Siberian pit viper), Gloydius shedaoensis (snake island pit viper), Gloydiusussuriensis (Ussuri pit viper), Hypnale (Knobby pit viper), Lachesis melanocephala (Blackhead bushmaster), Lachesis muta (Atlantic bushmaster), Ovophis okinavensis (Omehabu), Porthidium nasutum (Tropical rainforest pit viper), Porthidium ophryomegas (Slender pit viper), Protobothrops elegans (Sakishima pit viper), Protobothrops flavoviridis (Okinawa pit viper), Protobothrops mangshanensis (Manshan pit viper), Protobothrops mucrosquamatus (Taiwan pit viper), Protobothrops tokarensis (Tokara pit viper), Sistrurus catenatus (Masasogai), Sistrurus miliarius (Pygmy rattlesnake), Trimeresurus stejnegeri (Taiwanese Green Pit Viper), Trimeresurus albolabris (White-lipped Green Pit Viper), Trimeresurus erythrurus (Burmese Green Pit Viper), Trimeresurus gramineus (Green Pit Viper), Trimeresurus labialis (Nicobar Bamboo Pit Viper), Trimeresurus macrops (Large-eyed Pit Viper), Trimeresurus malabaricus (Malabar Pit Viper), Trimeresurus popeiorum (Pope Green Pit Viper), Trimeresurus purpureomaculatus (Mangrove Pit Viper), Trimeresurus sumatranus (Sumatran Pit Viper), Tropidolaemus wagleri (Wagler Pit Viper), Metlapilcoatlus mexicanus (Mexican Jumping Pit Viper), Metlapilcoatlus nummifer (Central American Jumping Pit Viper), Atheris squamigera (lizard bush viper), Bitis arietans (puff adder), Bitis Atropos (Cape mountain adder), Bitiscaudalis (horned puff adder), Bitis cornuta (western branched horned adder), Bitis gabonica (Central African Gabon viper), Bitis nasicornis (rhinoceros adder), Bitis parviocula (Ethiopian adder), Bitis rhinoceros (West African Gabon viper), Causus rhombeatus (common night adder), Cerastes (horned viper), Cerastes gasperettii (Arabian horned viper), Cerastes vipera (Saharan sand viper), Daboia palaestinae (Palestinian viper), Daboia russelii (Russell's viper), Daboia siamensis (Eastern Russell's viper), Daboia mauritanica (Moorish viper), Echis ocellatus (West African carpet viper), Echis carinatus (sawtooth viper), Echis coloratus (painted sawtooth viper), Echis pyramidum (Egyptian sawtooth viper), Macrovipera schweizeri (Milos viper), Macrovipera lebetinus (Levant viper), Montivipera bornmuelleri (Lebanese mountain viper), Montivipera latifii (Latific viper), Montivipera raddei (Armenian mountain viper), Montivipera xanthine (Ottoman viper), Pseudocerastes fieldi (Field horned viper), Pseudocerastes persicus (Persian horned viper), Vipera ammodytes (European horned viper), Vipera aspis (Asp viper), Vipera berus (European viper), Vipera latastei (Latast's viper), Vipera lotievi (Caucasian wild viper), Vipera seoanei (Basque viper), Vipera ursinii (Wild viper), DiadophisThe toxins are conjugated from one or more snake species selected from the group consisting of *Cranius punctatus* (collared snake), *Hydrodynastes gigas* (water cobra), *Hypsiglena torquata* (night snake), *Hypsiglena jani* (San Luis Toposi night snake), *Leptodeira ashmeadii*, *Philodryas olfersii* (Liechtenstein green racer), and combinations thereof. In one case, the venom injection comes from a sawtooth snake, water moccasin, lancehead, rattlesnake, Russell's viper, puff adder, or a combination thereof. In another case, the venom injection comes from a sawtooth snake, water moccasin, lancehead, rattlesnake, Russell's viper, and puff adder. In other cases, the venom injections were from the krait (Bungarus caeruleus), black mamba (Dendroaspis polylepsis), coastal taipan (Oxyuranus scutatellus), Cape cobra (Naja nivea), or a combination of these.

[0038] Antibodies or antigen-binding fragments that are broad-spectrum neutralizing antibodies against three-finger toxins (3FTx) are provided herein. In one embodiment, 3FTx is a long-chain neurotoxin. The antibody or antigen-binding fragment can neutralize alpha-bungarotoxin (krait), alpha-ellapitoxin (mamba), pseudonajatoxin (brown snake), alpha-cobratoxin (cobra), toxin B (king cobra), or a combination thereof. In one example, the antibody or antigen-binding fragment broadly neutralizes toxins from two, three, four, five or more of these long-chain neurotoxins. In a preferred embodiment, the antibody or antigen-binding fragment neutralizes the effect of the long-chain neurotoxin. In a preferred embodiment, the antibody or antigen-binding fragment conjugates one or more three-finger toxins in a manner that prevents the toxin from binding to human nicotinic acetylcholine receptors (nAChRs). In some embodiments, the antibody or antigen-binding fragment binds to at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the toxin-exposed area that would be filled when the toxin binds to nAChR. The antibody or antigen-binding fragment may be, for example, SNEURO_P01_D09. In one instance, SNEURO_P01_D09 includes VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 80, and VL CDR3 of SEQ ID NO: 101. In another instance, SNEURO_P01_D09 includes VH having the amino acid sequence of SEQ ID NO: 242, and VL having the amino acid sequence of SEQ ID NO: 273.

[0039] A method for identifying broad-spectrum neutralizing antibodies or antigen-binding fragments that selectively bind to two or more snake venom toxins belonging to a family of homologous antigens is provided herein, comprising the steps of (a) immunizing a subject with two or more homologous antigens; and (b) performing an iterative selection process to specifically identify cross-reactive antibodies or antigen-binding fragments from the B cell repertoire of the subject, wherein the iterative selection process down-selects possible candidates using two or more homologous antigens. The subject may be, for example, a mammal (e.g., human). In some embodiments, the family of homologous antigens includes, but is not limited to, three-finger toxins (3FTX), including long-chain neurotoxins. The iterative selection process includes immunoprecipitation of antibodies from serum obtained from the subject, fluorescence-activated cell sorting (FACS) of B cells obtained from the subject, panning of libraries derived from B cell RNA obtained from the subject presented in phages or yeast, or any combination thereof.

[0040] In some cases, subjects are immunized once or multiple times with multiple homologous antigens from the same family. In some cases, subjects are immunized with 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more different snake venom toxins. In some cases, the method further includes a step of screening the identified antibody or antigen-binding fragment for binding to multiple homologous antigens. The screening may be any suitable assay, including but not limited to affinity assays, kinetic assays, or combinations thereof. In some cases, the affinity assay or kinetic assay may include enzyme-linked immunosorbent assay (ELISA), Octet HTX assay, Biacore assay, or combinations thereof.

[0041] An antibody or antigen-binding fragment described herein that selectively binds to one or more snake toxins, comprising VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3, wherein (i) said VH CDR1 comprises the amino acid sequence FX2X3X4X5X6DX8H (where X2 is selected from N, T, and S; X3 is selected from F, L; X4 is selected from R, G, and S; X5 is selected from N and T; X6 is selected from Y, F, L; X8 is selected from M, I) (SEQ ID NO: 525); (ii) said VH CDR2 comprises the amino acid sequence X1X2X3X4X5X6X7GX9X 11 , 11 X 11 X 12 (where X1 is selected from P, S, and H; X2 is selected from V, G, and T; X3 is selected from V, F, L, and I; X4 is selected from D, G, and A; X5 is selected from Y, L, and H; X6 is selected from N, C, R, and T, and S; X7 is selected from V, F, and G; X9 is selected from A, E; X 10 is selected from Q, H; X 11 is selected from S, D, Y, and H; X 12 is selected from A and E) (SEQ ID NO: 526); (iii) said VH CDR3 comprises the amino acid sequence CX2RGTLYHYTSGSYX 15 SDAFDIW (where X2 is selected from V and A; X 15 is selected from Y and C) (SEQ ID NO: 527); (iv) said VL CDR1 comprises the amino acid sequence X1ASX4X5IX7X8X9LX 11 (where X1 is selected from Q and R; X4 is selected from Q and E; X5 is selected from D, G, T, and S; X7 is selected from R and S; X8 is selected from S, D, N, and I; X9 is selected from N, F, Y, W, and D; X 11(v) The VL CDR2 comprises the amino acid sequence X1ASX4X5X6X7 (wherein X1 is selected from G and A; X4 is selected from N, T and S; X5 is selected from L and S; X6 is selected from Q, L and E; X7 is selected from M and S) (Sequence ID 529); (vi) The VL CDR3 comprises the amino acid sequence CQQSYSTX8TF (wherein X8 is selected from I and H) (Sequence ID 530), or VL This specification provides an antibody or antigen-binding fragment in which CDR3 comprises the amino acid sequence CQQX4X5X6X7PX9TF (wherein X4 is selected from A and S; X5 is selected from N and Y; X6 is selected from I, T, and S; X7 is selected from P, F, and T; X9 is selected from P, Y, L, and W) (SEQ ID NO: 531).

[0042] Embedding by reference All published documents, patents, and patent applications referenced herein are incorporated by reference to the same extent as each individual published document, patent, or patent application is specifically and individually indicated as being incorporated by reference.

[0043] Novel features of the present invention are specifically described in the appended claims. A better understanding of the features and advantages of this disclosure will be obtained by referring to the subsequent detailed description illustrating explanatory embodiments in which the principles of the present invention are utilized, and to the appended drawings: [Brief explanation of the drawing]

[0044] [Figure 1A] Figure 1A provides an illustrative image of the SNEURO_P01_D09 light chain (D09LC) and SNEURO_P01_D09 heavy chain (D09HC) bound to toxin B.

[0045] [Figure 1B]Figure 1B provides an image diagram illustrating the α-bungarotoxin / nAChR complex (PDB:4UY2), in which the α9 ECD of the nicotinic acetylcholine receptor (nAChR) interacts with α-bungarotoxin in loop C.

[0046] [Figure 1C] Figure 1C provides an illustrative image of the footprint overlap between toxin B and D09 (dotted line) or nAChR (solid line).

[0047] [Figure 2] Figure 2 is a graph illustrating SNEURO_P01_D09 binding to Mamba-AVI-HIS at 0.8, 4, 20, 100, or 500 nM using Octect HTX BL1.

[0048] [Figure 3] Figure 3 is a graph illustrating SNEURO_P01_D09 binding to Taipan-AVI-HIS at 0.8, 4, 20, 100, or 500 nM using Octect HTX BL1.

[0049] [Figure 4] Figure 4 is a graph illustrating SNEURO_P01_D09 binding to Cobra-AVI-HIS at 0.8, 4, 20, 100, or 500 nM using Octect HTX BL1.

[0050] [Figure 5] Figure 5 is a graph illustrating SNEURO_P01_D09 binding to Mamba-AVI-HIS at 25°C at 0.8, 4, 20, or 100 nM. The KD was determined to be <74 pM.

[0051] [Figure 6] Figure 6 is a graph illustrating SNEURO_P01_D09 binding to Taipan-AVI-HIS at 25°C at 0.8, 4, 20, or 100 nM. The KD was determined to be 490 pM.

[0052] [Figure 7] Figure 7 is a graph illustrating SNEURO_P01_D09 binding to Cobra-AVI-HIS at 0.8, 4, or 20 nM at 25°C. The KD was determined to be <37 pM.

[0053] [Figure 8] Figure 8 is a graph illustrating SNEURO_P01_D09 binding to Mamba-AVI-HIS at 37°C at 0.8, 4, 20, or 100 nM. The KD was determined to be <57 pM.

[0054] [Figure 9] Figure 9 is a graph illustrating SNEURO_P01_D09 binding to Taipan-AVI-HIS at 37°C at 0.8, 4, or 20 nM. The KD was determined to be 960 pM.

[0055] [Figure 10] Figure 10 is a graph illustrating SNEURO_P01_D09 binding to Cobra-AVI-HIS at 37°C at 0.8, 4, or 20 nM. The KD was determined to be <37 pM.

[0056] [Figure 11] Figure 11 illustrates the production of universal antitoxins from toxins derived from just four snake venoms.

[0057] [Figure 12] Figure 12 illustrates the progress in the development of broad-spectrum monoclonal antibody candidates against alpha-neurotoxin, dendrotoxin, and PLA2.

[0058] [Figure 13A]Figures 13A–13F provide structures of the neurotoxin B / Centi-LNX-D9 Fab complex at 2.1 Å, illustrating the mechanism of neutralization. X-ray crystals of Centi-LNX-D9 against coastal taipan (Figure 13A), Cape cobra (Figure 13B), mamba (Figure 13C), and krait (Figure 13D) were obtained. The structures illustrate a broad-spectrum neutralization mechanism by molecular simulation of the native ligand epitope by VH CDR3 and supporting residues (Figure 13E). High-resolution epitope mapping (Figure 13F) provides the basis for the broad-spectrum neutralization capability of the Centi-LNX-D9 antibody observed from kinetics and in vivo defense. [Figure 13B] Figures 13A–13F provide structures of the neurotoxin B / Centi-LNX-D9 Fab complex at 2.1 Å, illustrating the mechanism of neutralization. X-ray crystals of Centi-LNX-D9 against coastal taipan (Figure 13A), Cape cobra (Figure 13B), mamba (Figure 13C), and krait (Figure 13D) were obtained. The structures illustrate a broad-spectrum neutralization mechanism by molecular simulation of the native ligand epitope by VH CDR3 and supporting residues (Figure 13E). High-resolution epitope mapping (Figure 13F) provides the basis for the broad-spectrum neutralization capability of the Centi-LNX-D9 antibody observed from kinetics and in vivo defense. [Figure 13C] Figures 13A–13F provide structures of the neurotoxin B / Centi-LNX-D9 Fab complex at 2.1 Å, illustrating the mechanism of neutralization. X-ray crystals of Centi-LNX-D9 against coastal taipan (Figure 13A), Cape cobra (Figure 13B), mamba (Figure 13C), and krait (Figure 13D) were obtained. The structures illustrate a broad-spectrum neutralization mechanism by molecular simulation of the native ligand epitope by VH CDR3 and supporting residues (Figure 13E). High-resolution epitope mapping (Figure 13F) provides the basis for the broad-spectrum neutralization capability of the Centi-LNX-D9 antibody observed from kinetics and in vivo defense. [Figure 13D]Figures 13A–13F provide structures of the neurotoxin B / Centi-LNX-D9 Fab complex at 2.1 Å, illustrating the mechanism of neutralization. X-ray crystals of Centi-LNX-D9 against coastal taipan (Figure 13A), Cape cobra (Figure 13B), mamba (Figure 13C), and krait (Figure 13D) were obtained. The structures illustrate a broad-spectrum neutralization mechanism by molecular simulation of the native ligand epitope by VH CDR3 and supporting residues (Figure 13E). High-resolution epitope mapping (Figure 13F) provides the basis for the broad-spectrum neutralization capability of the Centi-LNX-D9 antibody observed from kinetics and in vivo defense. [Figures 13E-13F] Figures 13A–13F provide structures of the neurotoxin B / Centi-LNX-D9 Fab complex at 2.1 Å, illustrating the mechanism of neutralization. X-ray crystals of Centi-LNX-D9 against coastal taipan (Figure 13A), Cape cobra (Figure 13B), mamba (Figure 13C), and krait (Figure 13D) were obtained. The structures illustrate a broad-spectrum neutralization mechanism by molecular simulation of the native ligand epitope by VH CDR3 and supporting residues (Figure 13E). High-resolution epitope mapping (Figure 13F) provides the basis for the broad-spectrum neutralization capability of the Centi-LNX-D9 antibody observed from kinetics and in vivo defense.

[0059] [Figure 14A-14B] Figures 14A–14B illustrate the results of recombinant toxin challenge in mice. Figure 14A provides data demonstrating that Centi-D09 (D9 in the figure) provided complete broad-area neutralizing protection against recombinant long-chain neurotoxins from black mamba, Cape cobra, coastal taipan, and indoor krait for 2 hours. Note: All lines for D9 treated animals overlap at 100% survival rate. Figure 14B provides data demonstrating complete protection lasting 25 hours against black mamba, coastal taipan, and Cape cobra.

[0060] [Figure 15]Figure 15 illustrates the toxin survival rate with bnAb Centi-D09. After obtaining LD100 for multiple toxins in mice, in vivo challenge studies demonstrated that Centi-D09 (Centi-D09 in the figure) provides complete broad-spectrum neutralization protection against all toxins from the Thai cobra (Naja kaouthia), black mamba (Dendroaspis polylepsis), and Cape cobra (Naja nivea). Note: All lines for D9 treated animals overlap at 100% survival rate.

[0061] [Figure 16] Figure 16 illustrates the ability of Centi-D09 to bind to and neutralize 3FTX snake venom toxins from diverse species (phylogeny shown in the left panel). Centi-D09 binds to 3FTX long-chain neurotoxins from all species tested (right panel, left column). Centi-D09 defended against in vivo challenges with isolated long-chain neurotoxins from various species (right panel, center column). Finally, Centi-D09 defended against all toxin challenges in vivo from diverse species, and in several cases where the defense was incomplete, its effect was shown to be synergistic with the PLA2 inhibitor valesprazib (right panel, right column). [Modes for carrying out the invention]

[0062] Detailed explanation Universal antitoxin compositions and methods for using universal antitoxin compositions to treat venomous snake bites are provided herein. A universal antitoxin composition may comprise one or more antibodies or antigen-binding fragments. The antibodies or antigen-binding fragments selectively bind to the snake toxins described herein and include VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3.

[0063] Typical VH CDR3 sequence The antibody or antigen-binding fragment may contain a VH CDR3 having an amino acid (AA) sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 1. [Table 1-1] [Table 1-2] [Table 1-3]

[0064] Typical VH CDR1 sequences The antibody or antigen-binding fragment may contain a VH CDR1 having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 2. [Table 2-1] [Table 2-2] [Table 2-3]

[0065] Typical VH CDR2 sequences The antibody or antigen-binding fragment may contain a VH CDR2 having an amino acid sequence that is at least 20%, 24%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 3. [Table 3-1] [Table 3-2] [Table 3-3]

[0066] Typical VL CDR1 sequences The antibody or antigen-binding fragment may contain a VL CDR1 having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 4. [Table 4-1] [Table 4-2]

[0067] Typical VL CDR2 sequences The antibody or antigen-binding fragment may contain a VL CDR2 having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 5. [Table 5-1] [Table 5-2]

[0068] Typical VL CDR3 sequences The antibody or antigen-binding fragment may contain a VL CDR3 having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 6. [Table 6-1] [Table 6-2] [Table 6-3]

[0069] Typical combinations of CDRs In one case, an antibody or antigen-binding fragment that selectively binds to a snaketoxin or combination of snaketoxins contains one of the following combinations of CDRs: [Table 7-1] [Table 7-2]

[0070] Antibodies or antigen-binding fragments can be optimized to increase their binding affinity. In one embodiment, such an antibody or antigen-binding fragment may have the heavy chain CDR combinations shown in Table 7A: [Table 7A]

[0071] Antibodies or antigen-binding fragments can be optimized to increase their binding affinity. In one embodiment, such an antibody or antigen-binding fragment may have the light chain CDR combinations shown in Table 7B: [Table 7B]

[0072] Typical FW-H1 array The antibody or antigen-binding fragment may contain a VH framework (FW)1 (FW-H1) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 8. [Table 8-1] [Table 8-2]

[0073] Typical FW-H2 array The antibody or antigen-binding fragment may contain a VH framework (FW)2 (FW-H2) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 9. [Table 9-1] [Table 9-2]

[0074] Typical FW-H3 arrangement The antibody or antigen-binding fragment may contain a VH framework (FW)3 (FW-H3) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 10. [Table 10-1] [Table 10-2]

[0075] Typical FW-H4 array The antibody or antigen-binding fragment may contain a VH framework (FW)4 (FW-H4) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 11. [Table 11-1] [Table 11-2]

[0076] Typical FW-L1 array The antibody or antigen-binding fragment may contain a VL framework (FW)1 (FW-L1) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 12. [Table 12-1] [Table 12-2]

[0077] Typical FW-L2 array The antibody or antigen-binding fragment may contain a VL framework (FW)2 (FW-L2) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 13. [Table 13-1] [Table 13-2]

[0078] Typical FW-L3 array The antibody or antigen-binding fragment may contain a VL framework (FW)3 (FW-L3) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 14. [Table 14-1] [Table 14-2]

[0079] Typical FW-L4 array The antibody or antigen-binding fragment may contain a VL framework (FW)4 (FW-L4) having an amino acid sequence that is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 15. [Table 15-1] [Table 15-2]

[0080] Typical VH sequence The antibody or antigen-binding fragment may contain a VH having an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 16. [Table 16-1] [Table 16-2] [Table 16-3] [Table 16-4] [Table 16-5] [Table 16-6] [Table 16-7] [Table 16-8]

[0081] Typical VL sequences The antibody or antigen-binding fragment may contain a VL having an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 100% identical to any one of the following sequences in Table 17. [Table 17-1] [Table 17-2] [Table 17-3] [Table 17-4] [Table 17-5] [Table 17-6]

[0082] Example antibody The antibodies or antigen-binding fragments described herein may include the variable heavy chain (VH) amino acid sequences and variable light chain (VL) amino acid sequences described herein.

[0083] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 224 and VL (PLA2_P01_A04) having the amino acid sequence of SEQ ID NO: 249.

[0084] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 225 and VL (PLA2_P01_A08) having the amino acid sequence of SEQ ID NO: 250.

[0085] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 226 and VL (PLA2_P01_B02) having the amino acid sequence of SEQ ID NO: 251.

[0086] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 227 and VL (PLA2_P01_B08) having the amino acid sequence of SEQ ID NO: 252.

[0087] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 228 and VL (PLA2_P01_C02) having the amino acid sequence of SEQ ID NO: 253.

[0088] The antibody or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 229 and VL (PLA2_P01_C03) having the amino acid sequence of SEQ ID NO: 254.

[0089] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 230 and VL (PLA2_P01_C04) having the amino acid sequence of SEQ ID NO: 255.

[0090] The antibody or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 231 and VL (PLA2_P01_D06.1) having the amino acid sequence of SEQ ID NO: 256.

[0091] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 231 and VL (PLA2_P01_D06.2) having the amino acid sequence of SEQ ID NO: 257.

[0092] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 232 and VL (PLA2_P01_G05) having the amino acid sequence of SEQ ID NO: 258.

[0093] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 233 and VL (PLA2_P01_G07) having the amino acid sequence of SEQ ID NO: 259.

[0094] The antibody or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 224 and VL (PLA2_P01_G12) having the amino acid sequence of SEQ ID NO: 249.

[0095] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 234 and VL (KUNITZ_P01_A03.1) having the amino acid sequence of SEQ ID NO: 260.

[0096] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 234 and VL (KUNITZ_P01_A03.2) having the amino acid sequence of SEQ ID NO: 261.

[0097] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 235 and VL (KUNITZ_P01_A07) having the amino acid sequence of SEQ ID NO: 262.

[0098] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 236 and VL (KUNITZ_P01_D07) having the amino acid sequence of SEQ ID NO: 263.

[0099] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 237 and VL (KUNITZ_P01_H05.1) having the amino acid sequence of SEQ ID NO: 264.

[0100] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 237 and VL (KUNITZ_P01_H05.2) having the amino acid sequence of SEQ ID NO: 265.

[0101] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 238 and VL (KUNITZ_P01_H07.1) having the amino acid sequence of SEQ ID NO: 266.

[0102] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 238 and VL (KUNITZ_P01_H07.2) having the amino acid sequence of SEQ ID NO: 267.

[0103] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 239 and VL (SNEURO_P01_A04) having the amino acid sequence of SEQ ID NO: 268.

[0104] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 240 and VL (SNEURO_P01_A08) having the amino acid sequence of SEQ ID NO: 269.

[0105] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 241 and VL (SNEURO_P01_A12) having the amino acid sequence of SEQ ID NO: 270.

[0106] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 239 and VL (SNEURO_P01_B05) having the amino acid sequence of SEQ ID NO: 271.

[0107] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 239 and VL (SNEURO_P01_B11) having the amino acid sequence of SEQ ID NO: 272.

[0108] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 242 and VL (SNEURO_P01_D09) having the amino acid sequence of SEQ ID NO: 273.

[0109] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 243 and VL (SNEURO_P01_E06) having the amino acid sequence of SEQ ID NO: 274.

[0110] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 243 and VL (SNEURO_P01_E09) having the amino acid sequence of SEQ ID NO: 275.

[0111] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 244 and VL (SNEURO_P01_F07) having the amino acid sequence of SEQ ID NO: 276.

[0112] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 239 and VL (SNEURO_P01_F09) having the amino acid sequence of SEQ ID NO: 277.

[0113] The antibody or antigen-binding fragments described herein include VH having the amino acid sequence of SEQ ID NO: 245 and VL (SNEURO_P01_H02) having the amino acid sequence of SEQ ID NO: 278.

[0114] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 246 and VL (VENM_M03_B12) having the amino acid sequence of SEQ ID NO: 279.

[0115] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 247 and VL (VENM_M12_G05) having the amino acid sequence of SEQ ID NO: 280.

[0116] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 248 and VL (VENM_M03_A06) having the amino acid sequence of SEQ ID NO: 281.

[0117] The antibodies or antigen-binding fragments described herein may include VH having the amino acid sequence of SEQ ID NO: 235 and VL (SNEURO_P01_H06) having the amino acid sequence of SEQ ID NO: 282.

[0118] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 429 and the VL of any one of SEQ ID NOs: 501 to 512.

[0119] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 430 and the VL of any one of SEQ ID NOs: 501 to 512.

[0120] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 431 and the VL of any one of SEQ ID NOs: 501 to 512.

[0121] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 432 and the VL of any one of SEQ ID NOs: 501 to 512.

[0122] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 433 and the VL of any one of SEQ ID NOs: 501 to 512.

[0123] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 434 and the VL of any one of SEQ ID NOs: 501 to 512.

[0124] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 435 and the VL of any one of SEQ ID NOs: 501 to 512.

[0125] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 436 and the VL of any one of SEQ ID NOs: 501 to 512.

[0126] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 437 and the VL of any one of SEQ ID NOs: 501 to 512.

[0127] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 438 and the VL of any one of SEQ ID NOs: 501 to 512.

[0128] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 439 and the VL of any one of SEQ ID NOs: 501-512.

[0129] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 440 and the VL of any one of SEQ ID NOs: 501 to 512.

[0130] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 441 and the VL of any one of SEQ ID NOs: 501 to 512.

[0131] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 442 and the VL of any one of SEQ ID NOs: 501 to 512.

[0132] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 443 and the VL of any one of SEQ ID NOs: 501 to 512.

[0133] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 444 and the VL of any one of SEQ ID NOs: 501 to 512.

[0134] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 445 and the VL of any one of SEQ ID NOs: 501 to 512.

[0135] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 446 and the VL of any one of SEQ ID NOs: 501 to 512.

[0136] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 447 and the VL of any one of SEQ ID NOs: 501 to 512.

[0137] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 448 and the VL of any one of SEQ ID NOs: 501 to 512.

[0138] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 449 and the VL of any one of SEQ ID NOs: 501 to 512.

[0139] The antibody or antigen-binding fragments described herein may include the VH of SEQ ID NO: 450 and the VL of any one of SEQ ID NOs: 501 to 512.

[0140] The antibody or antigen-binding fragments used herein may include VH of SEQ ID NO: 513 and VL of SEQ ID NO: 518. (SNEURO_P01_D09 VH Germline type)

[0141] The antibody or antigen-binding fragments used herein may include VH of SEQ ID NO: 514 and VL of SEQ ID NO: 519. (SNEURO_P01_D09 VK germline type)

[0142] The antibody or antigen-binding fragments used herein may include VH of SEQ ID NO: 515 and VL of SEQ ID NO: 520. (SNEURO_P01_D09 FW Germline Type)

[0143] The antibody or antigen-binding fragments used herein may include VH of SEQ ID NO: 516 and VL of SEQ ID NO: 521. (SNEURO_P01_D09 VK L3 NS->QS)

[0144] The antibody or antigen-binding fragments used herein may include VH of SEQ ID NO: 517 and VL of SEQ ID NO: 522. (SNEURO_P01_D09 VK L3 NS->NT)

[0145] The antibody or antigen-binding fragments used herein may include VH of SEQ ID NO: 523 and VL of SEQ ID NO: 524. (Centi-DTX-B03)

[0146] In one embodiment, the antibody or antigen-binding fragments herein comprise VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3, wherein (i) VH CDR1 comprises the amino acid sequence FX2X3X4X5X6DX8H (wherein X2 is selected from N, T, and S; X3 is selected from F, L; X4 is selected from R, G, and S; X5 is selected from N and T; X6 is selected from Y, F, and L; X8 is selected from M, I) (SEQ ID NO: 525); and (ii) VH CDR2 comprises the amino acid sequence X1X2X3X4X5X6X7GX9X 10 X 11 X 12(In the formula, X1 is selected from P, S, and H; X2 is selected from V, G, and T; X3 is selected from V, F, L, and I; X4 is selected from D, G, and A; X5 is selected from Y, L, and H; X6 is selected from N, C, R, T, and S; X7 is selected from V, F, and G; X9 is selected from A and E; X 10 is selected from Q, H; X 11 is selected from S, D, Y, and H; X 12 (iii)VH CDR3 is selected from A and E) (Sequence ID 526) and includes; (iii)VH CDR3 is amino acid sequence CX2RGTLYHYTSGSYX 15 SDAFDIW(wherein X2 is selected from V and A; X 15 (iv)VL CDR1 contains (selected from Y and C) (SEQ ID NO: 527); (iv)VL CDR1 has the amino acid sequence X1ASX4X5IX7X8X9LX 11 (In the formula, X1 is selected from Q and R; X4 is selected from Q and E; X5 is selected from D, G, T, and S; X7 is selected from R and S; X8 is selected from S, D, N, and I; X9 is selected from N, F, Y, W, and D; X 11 (v) VL CDR2 contains the amino acid sequence X1ASX4X5X6X7 (wherein X1 is selected from G and A; X4 is selected from N, T, and S; X5 is selected from L and S; X6 is selected from Q, L, and E; X7 is selected from M and S) (Sequence ID 529); (vi) VL CDR3 contains the amino acid sequence CQQSYSTX8TF (wherein X8 is selected from I and H) (Sequence ID 530), or VL CDR3 contains the amino acid sequence CQQX4X5X6X7PX9TF (wherein X4 is selected from A and S; X5 is selected from N and Y; X6 is selected from I, T, and S; X7 is selected from P, F, and T; X9 is selected from P, Y, L, and W) (SEQ ID NO: 531).

[0147] In some embodiments, the antibody or antigen-binding fragment of the present invention specifically binds to multiple homologous members of the three-finger toxin (3FTX) family found in Elapidae. The antibody or antigen-binding fragment may specifically bind to one or more homologous members of the 3FTX family that are alpha-neurotoxins. The antibody or antigen-binding fragment may specifically bind to one or more homologous members of the 3FTX family that are long-chain neurotoxins. In certain embodiments, the antibody or antigen-binding fragment neutralizes the effect of the long-chain neurotoxin. In certain embodiments, the antibody or antigen-binding fragment binds to one or more three-finger toxins in a manner that prevents the toxin from binding to the human nicotinic acetylcholine receptor (nAChR). In some embodiments, the antibody or antigen-binding fragment binds to at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the toxin exposure area that would be filled if the toxin bound to the nAChR.

[0148] In certain embodiments, the antibody or antigen-binding fragment binds to two or more neurotoxin members of the 3FTX family using the same multispecific paratope (i.e., a single set of CDRs capable of binding to two or more long-chain neurotoxin members of the 3FTX family).

[0149] In some embodiments, the antibody or antigen-binding fragment specifically binds to a member of the long-chain neurotoxin 3FTX family to which the highly immunized subject from whom the antibody was collected has never been exposed.

[0150] Antibodies or antigen-binding fragments may specifically bind to multiple members of the long-chain neurotoxin 3FTX family. Antibodies or antigen-binding fragments may specifically bind to two three-finger toxins sharing less than 90%, 80%, 70%, 60%, or 50% sequence identity. Antibodies or antigen-binding fragments may specifically bind to three non-paired three-finger toxins sharing more than 90%, 80%, 70%, 60%, or 50% sequence identity. Antibodies or antigen-binding fragments may specifically bind to four, five, or six non-paired three-finger toxins sharing more than 90%, 80%, 70%, 60%, or 50% sequence identity.

[0151] In one embodiment, the antibody or antigen-binding fragment binds to alpha-bungarotoxin found in the Indian krait, alpha-ellapitoxin found in the black mamba, and alpha-neurotoxin found in the Thai cobra. In one embodiment, the antibody or antigen-binding fragment may bind to long-chain and short-chain neurotoxins of the 3FTX family.

[0152] In one embodiment, the antibody or antigen-binding fragment binds to two, three or more of the following long-chain neurotoxin three-finger toxins: long-chain neurotoxin three-finger toxins found in mamba species (genus dendroaspis), cobra species (genus naja), taipan species (genus oxyuranus), and krait species (genus bungarus). In one embodiment, the antibody or antigen-binding fragment binds to long-chain neurotoxin three-finger toxins found in at least one mamba species, at least one cobra species, at least one taipan species, and at least one krait species. In one embodiment, the antibody or antigen-binding fragment binds to long-chain neurotoxin three-finger type toxins from two, three or more of the following species: black mamba (dendroaspis polylepis), Thai cobra (naja kaouthia), coastal taipan (oxyuranus scutellatus), and indoor krait (bungarus caeruleus). In a particular embodiment, the antibody or antigen-binding fragment binds to long-chain neurotoxin three-finger type toxins from black mamba (dendroaspis polylepis), Thai cobra (naja kaouthia), coastal taipan (oxyuranus scutellatus), and indoor krait (bungarus caeruleus).

[0153] In one embodiment, the antibody or antigen-binding fragment binds to long-chain neurotoxins from two or more species of cobra (in the genus Naja). In another embodiment, the antibody or antigen-binding fragment binds to long-chain neurotoxins from two or more species of cobra found on different continents. The antibody or antigen-binding fragment may bind to 3FTX found in Asian snake species and 3FTX found in African snake species. For example, the antibody or antigen-binding fragment may bind to the Indian cobra (Naja naja) and the Egyptian cobra (Naja haje). The antibody or antigen-binding fragment may bind to 3FTX found in cobras found in India, Africa, and Indonesia.

[0154] In one embodiment, the antibody or antigen-binding fragment binds to long-chain neurotoxins from one or more species of the Elapidae family found in Australia, one or more species of the Elapidae family found in India, and one or more species of the Elapidae family found in Africa. For example, the antibody or antigen-binding fragment may bind to long-chain neurotoxins found in coastal taipan (Australia), Indian cobra (India), and black mamba (Africa).

[0155] The antibody or antigen-binding fragment may bind to at least one long-chain neurotoxin found in cobras of the genus Naja and at least one long-chain neurotoxin found in cobras of the genus Ophiophagus.

[0156] In one embodiment, the antibody or antigen-binding fragment binds to at least one long-chain neurotoxin found in the toxins of snakes of the family Elapidae but not in the toxins of snakes of the subfamily Hydrophiinae, and to at least one long-chain neurotoxin found in the toxins of snakes of the subfamily Hydrophiinae. In one embodiment, the antibody or antigen-binding fragment binds to a long-chain neurotoxin from the blue-spotted sea snake (Laticauda colubrina) and a long-chain neurotoxin from at least one species of the family Elapidae. In one embodiment, the antibody or antigen-binding fragment binds to two or more of the following long-chain neurotoxins: long-chain neurotoxins found in the species aipysurus, the species hydrophis, and the species laticauda.

[0157] A combination or group of antibodies or antigen-binding fragments as described herein may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more different antibodies. In one example, a group of antibodies or antigen-binding fragments as described herein may include one antibody or antigen-binding fragment as described herein. In one example, a combination or group of antibodies or antigen-binding fragments as described herein may include two different antibodies or antigen-binding fragments. In one example, a combination or group of antibodies or antigen-binding fragments as described herein may include three different antibodies or antigen-binding fragments. In one example, a combination or group of antibodies or antigen-binding fragments as described herein may include four different antibodies or antigen-binding fragments. In one example, a combination or group of antibodies or antigen-binding fragments as described herein may include five different antibodies or antigen-binding fragments. In one example, a combination or group of antibodies or antigen-binding fragments as described herein may include six different antibodies or antigen-binding fragments. In one example, a combination or group of antibodies or antigen-binding fragments as described herein may include seven different antibodies or antigen-binding fragments. In another example, a combination or group of antibodies or antigen-binding fragments as described herein may include eight different antibodies or antigen-binding fragments. In yet another example, a combination or group of antibodies or antigen-binding fragments as described herein may include nine different antibodies or antigen-binding fragments. In yet another example, a combination or group of antibodies or antigen-binding fragments as described herein may include ten different antibodies or antigen-binding fragments. In yet another example, a combination or group of antibodies or antigen-binding fragments as described herein may include more than ten different antibodies or antigen-binding fragments. A combination or group of antibodies or antigen-binding fragments as described herein may include antibodies that bind to snakes from continents, countries, or regions. For example, a combination or group of antibodies or antigen-binding fragments as described herein may be useful for treating snake bites in North, Central America, South America, the Middle East, India, China, Africa, Asia, Australia, and other regions.Alternatively, combinations or populations of antibodies or antigen-binding fragments as described herein may be useful for treating snakebites in a region, for example, the northeastern United States, the southeastern United States, the southwestern United States, or the northwestern United States. Such combinations or populations of antibodies or antigen-binding fragments as described herein are useful when the bitten subject cannot be identified as having been bitten by a specific snake.

[0158] This application also discloses a method for identifying broad-spectrum neutralizing antibodies against a family of homologous antigens (e.g., snake venom toxins). Families of homologous antigens found in snake venom include three-finger toxins (3FTX), long-chain neurotoxins, short-chain neurotoxins, snake venom metalloproteinases (SVMPs), snake venom serine proteases (SVSPs), and phospholipase A2 (PLA2). A family of homologous antigens may be another toxin or a family of toxins present in multiple snake species.

[0159] The method involves immunization of a human or another animal ("highly immunized subject") with two or more homologous antigens. In some embodiments, the method may include immunization with 2, 3, 4, 5, 6, 7, 8, 9, or 10 homologous antigens. The method may include immunization with between 10 and 20 homologous antigens. The method may include immunization with between 20 and 30 homologous antigens. The method may include immunization with between 30 and 40 homologous antigens. In some embodiments, the immunization may include recombinant homologous antigens. These antigens can be selected to sample a diverse subset of homologous antigens. In some embodiments, the immunization may include whole toxins from different species containing homologous toxin antigens. These species can be selected to be phylogenetically diverse.

[0160] Each of these immunizations can be repeated two or more times. In some embodiments, the method may include immunizations repeated two, three, four, five, six, seven, eight, nine, or ten times. The method may include immunizations repeated between 10 and 20 times. The method may include immunizations repeated between 20 and 30 times. The method may include immunizations repeated between 30 and 40 times. The method may include immunizations repeated more than 40 times. For example, an immunization schedule involving three homologous antigens repeated twice may be: immunization with antigen A1, then A2, then A3, then again with A1, A2, and A3.

[0161] The method may further include the identification of monoclonal antibodies from subjects exhibiting cross-reactivity to two or more homologous antigens. In certain embodiments, the method includes repeated selection rounds to identify such cross-reactive monoclonal antibodies. Examples of such methods for selection include immunoprecipitation of antibodies from serum; fluorescence-activated cell sorting (FACS) of B cells; or panning of libraries presented in phages or yeast.

[0162] Regardless of the specific selection technique used, the method of the present invention involves repeated selection using multiple homologous antigens to specifically enrich antibodies isolated from a highly immunized subject that are cross-reactive and broadly neutralizing to multiple members of a homologous antigen family. The repeated selection round may utilize sequences of different homologous antigens to preferentially select cross-reactive antibodies. The repeated selection round may utilize members of the homologous antigen family from which the highly immunized subject has not been immunized to preferentially select cross-reactive antibodies. The selection process may utilize the use of 2, 3, 4, 5, 6, 7, 8, 9, or 10 different members of the homologous antigen family. Clones identified using repeated down-selection are guaranteed to be cross-reactive to all members of the homologous antigen family included in the repeated down-selection method.

[0163] Exemplary methods for identifying cross-reactive antibodies using repeated selection with phage display are given herein. The method may include isolation of PBMCs or B cells from the highly immunized subject. The method may further include RNA extraction. The method may further include reverse transcription of cDNA from the extracted RNA. The method may further include amplification (e.g., by PCR) of the antibody variable domain. The method may further include construction of a DNA library of the antibody variable domain. The method may further include digestion of the DNA library and cloning into a phage display vector. The method may further include panning to one or more toxins. The method may include panning to 2, 3, 4, 5, 6, 7, 8, 9, or 10 different members of a homologous antigen family, so that the retained clones should be cross-reactive to all members utilized in the repeated panning process. The toxins may include homologous toxins to which the highly immunized subject has been previously exposed, in addition to homologous toxins to which the highly immunized subject has not been previously exposed. The method may further include screening antibody clones for binding, for example, by ELISA or kinetics (e.g., Octet HTX or Biacore).

[0164] Modified antibody This disclosure provides modified antibodies. Modified antibodies may include antibodies having one or more modifications that can improve their activity, binding, specificity, selectivity, or other characteristics. In one embodiment, this disclosure provides modified antibodies (which may include heteromultimers) comprising antibodies described herein. References to modified antibodies herein also refer to modified antigen-binding fragments. Modified antibodies may include bispecific modified antibodies or antigen-binding fragments, triplicate modified antibodies or antigen-binding fragments, or quadruplespecific modified antibodies or antigen-binding fragments.

[0165] Modified antibodies may include human modified antibodies. Amino acid sequence variants of modified antibodies, which can be prepared by introducing appropriate nucleotide changes into modified antibody DNA or by synthesizing a desired modified antibody polypeptide, are also included herein. Such variants include, for example, deletions of residues from the amino acid sequences of first and second polypeptides forming the modified antibody, insertions of residues into the amino acid sequence, and / or substitutions of residues within the amino acid sequence. Any combination of deletions, insertions, and substitutions is performed to reach the final construct, provided that the final construct has the desired antigen-binding properties. Amino acid changes can also alter the post-translational processes of the modified antibody, such as changing the number or location of glycosylation sites.

[0166] "Alanine scanning mutagenesis" may be a useful method for identifying specific residues or regions of a modified antibody polypeptide that may be favorable sites for mutagenesis. Here, one or a group of target residues is identified (e.g., charged residues such as Arg, Asp, His, Lys, and Glu), and replaced with a neutral or charged amino acid (e.g., alanine or polyalanine) to affect the interaction between the amino acid and the surrounding aqueous environment inside or outside the cell. The domain demonstrating the sensitivity of the functional group to the substitution is then refined by introducing further or other variants to or from the substitution site. Thus, the site for introducing amino acid sequence mutations is predetermined, but the nature of the mutation itself does not need to be predetermined.

[0167] Typically, substitutions may involve conserved amino acid substitutions in the non-functional region of the modified antibody. Exemplary substitutions are shown below. [Table 18]

[0168] Covalent modification of antibodies, antigen-binding fragments, or modified antibody polypeptides is included within the scope of the present disclosure. Covalent modification of a modified antibody can be introduced into the molecule by reacting a target amino acid residue of the modified antibody or fragment thereof with an organic derivatizing agent capable of reacting with a selected side chain or N- or C-terminal residue. Another type of covalent modification of a modified antibody polypeptide can involve changing the native glycosylation pattern of the polypeptide. As used herein, "changing" can mean deleting one or more sugar chain moieties found in the original modified antibody and / or adding one or more glycosylation sites not present in the original modified antibody. Addition of a glycosylation site to a modified antibody polypeptide can be accomplished by changing the amino acid sequence such that it contains one or more N-linked glycosylation sites. Changes can also be made by addition of one or more serine or threonine residues (to O-linked glycosylation sites) to the original modified antibody sequence or by substitution with said residues. Briefly, the modified antibody amino acid sequence can be changed by changes at the DNA level, specifically by mutating preselected bases of the DNA encoding the modified antibody polypeptide such that codons that will translate to the desired amino acids are generated. Another means of increasing the number of sugar chain moieties on a modified antibody polypeptide is by chemical or enzymatic coupling of a glycoside to the polypeptide. Removal of sugar chain moieties present on a modified antibody can be accomplished chemically or enzymatically.

[0169] Another type of covalent modification of a modified antibody involves linking the modified antibody polypeptide to one of a variety of non-proteinaceous polymers, such as polyethylene glycol, polypropylene glycol, or polyoxyalkylene.

[0170] Methods for complexing a binder or an antibody or an antigen-binding fragment thereof with another agent are known in the art. Such methods can utilize one of several available heterobifunctional reagents used to couple or link molecules.

[0171] In one instance, the Fc portion of an antibody can be modified to increase the half-life of a molecule that circulates in the blood when administered to a subject.

[0172] In addition, antibodies can be produced or expressed such that they do not contain fucose on their complex N-glycosidic-linked sugar chains to increase their effector function. Similarly, antibodies can be attached at the C-terminal side of all or part of the immunoglobulin heavy chains derived from any antibody isotype, such as IgG, IgA, IgE, IgD, and IgM, and isotype subclasses, such as IgG1, IgG2b, IgG2a, IgG3, and IgG4.

[0173] Glycosylation of immunoglobulins has been shown to have a significant impact on their effector function, structural stability, and secretion rate from antibody-producing cells. Antibodies or antigen-binding fragments herein can be glycosylated. Glycosylation at variable domain framework residues can alter the binding interaction between an antibody and an antigen. The present disclosure includes criteria for selecting a limited number of amino acids within the framework or CDR of an immunoglobulin chain to mutate (e.g., by residue substitution, deletion, and / or addition) to increase the affinity of an antibody.

[0174] Linkers for conjugating an antibody to another moiety are within the scope of the present disclosure. Association (binding) of an antibody with a label includes, but is not limited to, covalent and non-covalent interactions, chemical conjugation, and recombinant techniques.

[0175] Antibodies, or their antigen-binding fragments, can be modified for various purposes, for example, by the addition of polyethylene glycol (PEG). PEG modification (PEGylation) can result in one or more of the following: improved circulation time, improved solubility, improved resistance to proteolysis, reduced antigenicity and immunogenicity, improved bioavailability, reduced toxicity, improved stability, and easier formulation.

[0176] Antibodies or antigen-binding fragments can be conjugated to affinity tags (e.g., purification tags), or recombination operations can be performed using such tags. For example, affinity tags such as the His6 tag (His-His-His-His-His-His) (SEQ ID NO: 283) are described.

[0177] Since predicting the properties of variant-modified antibodies in advance is often difficult, it will be understood that some screening of recovered variants may be necessary to select the optimal variant. An exemplary screening method for recovered variants is described in the examples below.

[0178] Methods for expressing antibodies Methods for producing either these antibodies or polypeptides are also provided herein. Polypeptides can be produced by proteolytic or other degradation of antibodies, by recombinant methods as described above (i.e., single or fusion polypeptides), or by chemical synthesis. Antibody polypeptides, in particular shorter polypeptides of up to approximately 50 amino acids, can be produced by chemical synthesis. Methods for chemical synthesis are commercially available. For example, antibodies can be produced using a solid-phase method with an automated polypeptide synthesizer.

[0179] Antibodies can be produced by recombination, which involves first isolating the antibody and antibody-producing cells from a host animal, obtaining the gene sequence, and then recombinantly expressing the antibody in host cells (e.g., CHO cells) using that gene sequence. Another possible method involves expressing the antibody sequence in plants (e.g., tobacco) or transgenic milk. Methods for recombinantly expressing antibodies in plants or milk are disclosed. Methods for producing antibody derivatives, such as single chains, are also within the scope of this disclosure.

[0180] As used herein, “host cell” includes individual cells or cell cultures that may or may have been recipients of a vector for the incorporation of a polynucleotide insert. A host cell includes offspring of a single host cell, which may not necessarily be completely identical (in terms of morphology or genomic DNA complement) to the original parent cell due to spontaneous, accidental, or intentional mutations. A host cell includes cells transfected with the polynucleotides of this disclosure.

[0181] Antibody-encoding DNA can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that can specifically bind to the genes encoding the heavy and light chains of monoclonal antibodies). Hybridoma cells can serve as a source of such DNA. Once isolated, the DNA can be placed in one or more expression vectors (e.g., the expression vectors disclosed in PCT Publication No. WO87 / 04462), and these can then be transfected into host cells that would otherwise not produce immunoglobulin proteins, such as E. coli cells, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells, to achieve the synthesis of monoclonal antibodies in recombinant host cells. The DNA can also be modified, for example, by substituting the coding sequences of human heavy and light chain constant domains for homologous mouse sequences, or by covalently linking all or part of the coding sequence of a non-immunoglobulin polypeptide to the immunoglobulin coding sequence. In this way, “chimeric” or “hybrid” antibodies having the antibody binding specificity described herein are prepared.

[0182] Vectors encoding one or more antibodies or antigen-binding fragments are contemplated herein. As used herein, “vector” means a construct capable of delivering and possibly expressing one or more genes or sequences of interest in a host cell. Examples of vectors include, but are not limited to, viral vectors; naked DNA or RNA expression vectors; plasmids, cosmids, or phage vectors; DNA or RNA expression vectors with cationic condensants; liposome-encapsulated DNA or RNA expression vectors; and certain eukaryotic cells, e.g., producing cells.

[0183] As used herein, “expression regulatory sequence” means a nucleic acid sequence that directs the transcription of a nucleic acid. An expression regulatory sequence may be a promoter, such as a constitutive or inductive promoter, or an enhancer. The expression regulatory sequence is operably ligated to the nucleic acid sequence to be transcribed. An expression vector can be used to direct the expression of an antibody. An expression vector can be administered to achieve the expression of an exogenous protein in vivo.

[0184] A widely used mammalian expression system for high-level production utilizes Lonza's GS Gene Expression System (trademark). This system uses a viral promoter and glutamate metabolism-based selection to generate stable mammalian cell lines in high yield.

[0185] A widely used mammalian expression system for alternative high-level production utilizes gene amplification in dihydrofolate reductase-deficient ("dhfr") Chinese hamster ovary cells. This system is based on the dihydrofolate reductase "dhfr" gene, which encodes the DHFR enzyme that catalyzes the conversion of dihydrofolate to tetrahydrofolate. To achieve high production, dhfr-CHO cells are transfected with an expression vector containing the functional DHFR gene along with the gene encoding the desired protein. In this case, the desired protein is a recombinant antibody heavy chain and / or light chain.

[0186] By increasing the amount of the competitive DHFR inhibitor methotrexate (MTX), recombinant cells develop resistance by amplifying the dhfr gene. Under typical circumstances, the amplification units utilized are much larger than the size of the dhfr gene, resulting in co-amplification of the antibody heavy chain.

[0187] When large-scale production of proteins such as antibody chains is desired, both the expression level and stability of the cells that will be used must be considered.

[0188] This application provides one or more isolated polynucleotides (nucleic acids) encoding an antibody or antigen-binding fragment herein, vectors containing such polynucleotides, and host cells and expression systems for transcribing and translating such polynucleotides into polypeptides.

[0189] This application also provides constructs in the form of plasmids, vectors, transcription or expression cassettes that include at least one of the above polynucleotides.

[0190] This application also provides recombinant host cells containing one or more of the above constructs. Nucleic acids encoding any antibody or antigen-binding fragment herein form aspects of this application, and methods of producing an antibody also form aspects of this application, which methods include expression from the encoding nucleic acid therefrom. Expression can be achieved by culturing a recombinant host cell containing the nucleic acid under suitable conditions. After production by expression, the antibody or a portion thereof can be isolated and / or purified using any suitable technique and then used as appropriate.

[0191] Systems for cloning and expression of polypeptides in a variety of different host cells are contemplated herein for use.

[0192] Further aspects provide host cells containing the nucleic acids herein using any suitable method. Still further aspects provide methods that include the step of introducing such nucleic acids into a host cell. Following introduction, expression from the nucleic acid can be caused or enabled, for example, by culturing the host cell under conditions for expression of the gene.

[0193] One or more polynucleotides encoding an antibody or antigen-binding fragment can be prepared by recombination / synthetic means, in addition to cloning, or without cloning. In further embodiments, the recombinant DNA molecule of the antibody or antigen-binding fragment described herein, or the entire DNA sequence of the cloned gene, can be operably ligated to an expression control sequence that can be introduced into a suitable host using any preferred method.

[0194] Nucleic acid sequences can be expressed by operably ligating them to expression regulatory sequences within an appropriate expression vector, and by transforming appropriate host cells using that expression vector. Nucleic acid sequences can be expressed using any of the wide variety of expression regulatory sequences—sequences that control the expression of the nucleic acid sequences to which they are operably ligated—in these vectors.

[0195] A wide variety of host / expression vector combinations can be used to express the nucleic acid sequences of this disclosure. It will be understood that not all vectors, expression regulatory sequences, and hosts will function equally well to express the nucleic acid sequences. Not all hosts will function equally well in the same expression system. In some embodiments, when selecting a vector, the host is considered in such a way that the vector can function within it. The copy number of the vector, its ability to control its copy number, and the expression of any other proteins encoded by the vector, such as antibiotic markers, may also be considered. In some particular embodiments, when selecting a vector, the host is considered in such a way that the vector can function within it. The copy number of the vector, its ability to control its copy number, and the expression of any other proteins encoded by the vector, such as antibiotic markers, may also be considered.

[0196] The present invention also provides a method that includes using the above-described construct in an expression system to express the above-described antibody (or a portion thereof). Considering these and other factors, various vector / expression regulatory sequence / host combinations can be constructed that can express nucleic acid sequences using fermentation or in large-scale animal cultures.

[0197] The simultaneous incorporation of nucleic acids encoding antibodies (or a portion thereof) and changes in selected amino acid positions can be achieved by various suitable methods, such as recombination and chemical synthesis.

[0198] A method for expressing an antibody or antigen-binding fragment (e.g., an antibody or antigen-binding fragment) in a subject is provided herein, comprising the step of administering to the subject a composition comprising a polynucleotide (e.g., mRNA) encoding the antibody or antigen-binding fragment.

[0199] In some cases, the steps for administering polynucleotides may include enteral, gastrointestinal, oral, transdermal, intracutaneous, intradermal, subcutaneous, nasal, intravenous, intraperitoneal, intra-arterial, intramuscular, intraosseous, mucosal, aerated, or sublingual administration. In some cases, polynucleotides may be administered by one or more routes.

[0200] Antibodies or antigen-binding fragments can be synthesized in a target based at least in part on a polynucleotide encoding the antibody or antigen-binding fragment. For example, the polynucleotide can enter the target cell, and the antibody or antigen-binding fragment can be synthesized at least in part using the target cell's transcription and / or translation mechanisms. In some cases, for example, if the polynucleotide is an mRNA molecule, the antibody or antigen-binding fragment can be synthesized at least in part using the target cell's translation mechanisms (e.g., ribosomes, tRNA, etc.). In some cases, the antibody or antigen-binding fragment can be transported from the cell to the target plasma after translation.

[0201] composition Compositions comprising antibodies or antigen-binding fragments as described herein can be prepared for storage in the form of lyophilized formulations or aqueous solutions by mixing antibodies or antigen-binding fragments of desired purity with pharmaceutically acceptable carriers, excipients, or stabilizers as needed (Remington, The Science and Practice of Pharmacy 20th Ed. Mack Publishing (2000)).

[0202] As used herein, “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” includes any material that, when combined with the active ingredient, enables the ingredient to retain its biological activity and is non-reactive with the target immune system. Examples include, but are not limited to, any of the standard pharmaceutical carriers, e.g., phosphate-buffered saline solution, water, emulsions, e.g., oil / water emulsions, and various types of wetting agents. Preferred diluents for aerosol or parenteral administration are phosphate-buffered saline or physiological (0.9%) saline. Compositions containing such carriers are formulated by well-known conventional methods (see, for example, Remington's Pharmaceutical Sciences, 18th edition, A. Gennaro, Ed., Mack Publishing Co., Easton, Pa., 1990; and Remington, The Science and Practice of Pharmacy 20th Ed. Mack Publishing, 2000).

[0203] Acceptable carriers, excipients, or stabilizers are non-toxic to the recipient at the dosage and concentration used, and include buffers, e.g., phosphates, citrates, and other organic acids; salts, e.g., sodium chloride; antioxidants, including ascorbic acid and methionine; preservatives (e.g., octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkylparabens, e.g., methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); low molecular weight (less than approximately 10 residues) polypeptides; proteins, e.g. The materials may also include serum albumin, gelatin, or immunoglobulin; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents, e.g., EDTA; sugars, e.g., sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, e.g., sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, e.g., TWEEN®, PLURONICS®, or polyethylene glycol (PEG).

[0204] The compositions to be used for in vivo administration may be sterilized. This can be achieved, for example, by filtration with a sterile filtration membrane or by any other sterilization method recognized in the art. The antibody composition is generally placed in a container having a sterile outlet, such as an intravenous solution bag or vial with a stopper that can be punctured with a subcutaneous needle. Other methods for sterilization and filtration are known in the art and are contemplated herein.

[0205] In some embodiments of this disclosure, compositions are formulated to be pyrogen-free so that they are acceptable for administration to a subject.

[0206] The compositions according to this disclosure may be in unit dosage forms for intravenous, oral, parenteral, or rectal administration, or administration by inhalation or inhalation, such as solutions or suspensions, tablets, pills, capsules, powders, granules, or suppositories.

[0207] The phrase "pharmaceutically acceptable" refers to molecular entities and compositions that, when administered to a subject, are physiologically tolerable and do not typically cause allergic reactions or similar adverse reactions, such as stomach upset or dizziness.

[0208] In some cases, antibodies or antigen-binding fragments can be conjugated to one or more carriers. The carriers may be active and / or inactive. Examples of well-known carriers include polypropylene, polystyrene, polyethylene, dextran, nylon, amylase, glass, natural and modified cellulose, polyacrylamide, agarose, and magnetite. For the purposes of this disclosure, the properties of the carriers may be either soluble or insoluble. Those skilled in the art will know of, or can determine, other carriers suitable for antibody conjugation, for example, by using routine experiments.

[0209] Various embodiments envision the use of antibodies or antigen-binding fragments to manufacture pharmaceuticals for treating the conditions, diseases, or disorders described herein. Pharmaceuticals can be formulated based on the physical characteristics of the subject requiring treatment, and can be formulated into single or multiple formulations based on the stage of the condition, disease, or disorder. Pharmaceuticals can be packaged in suitable packaging with appropriate labeling for distribution to hospitals and clinics, the labeling being for the treatment of subjects having the diseases described herein. Pharmaceuticals can be packaged as a single unit or as multiple units. Instructions for use regarding the dosage and administration of the composition can be included with the package as described below. This disclosure further relates to pharmaceuticals of antibodies or antigen-binding fragments and pharmaceutically acceptable carriers.

[0210] kit Kits comprising one or more antibodies or antigen-binding fragments are provided herein. Container means comprising one or more antibodies or antigen-binding fragments are also provided herein. The container means may include, but are not limited to, vials, syringes, bottles, intravenous (IV) bags, ampoules, or any other suitable containers, and may be any suitable container capable of holding a liquid or lyophilized composition. Syringes may hold any volume of a liquid suitable for injection into a subject, including but not limited to 0.5 cc, 1 cc, 2 cc, 5 cc, 10 cc or more. In some embodiments, the antibodies or antigen-binding fragments are lyophilized, and the kit includes one or more suitable buffers for reconstitution before injection.

[0211] The kit may include one or more instructions for use describing the use of one or more antibodies or antigen-binding fragments. The kit may include one or more labels describing the contents and use of one or more antibodies. Treatment method

[0212] This disclosure provides a method for preventing or treating a subject suffering from poison injection, comprising administering an antibody or antigen-binding fragment as defined herein to the subject. In one case, the subject to be treated exhibits symptoms prior to the administration of the antibody. In another case, the subject to be treated does not exhibit symptoms prior to the administration of the antibody. "Subject" as used herein includes, but is not limited to, humans, rodents, primates, etc. The treatment may result in partial or complete treatment of one or more symptoms of poison injection.

[0213] The antibodies or antigen-binding fragments described herein may be administered to a subject in an amount that achieves at least partial or complete relief of one or more symptoms. Relief may be, for example, a reduction of about 5% or more of one or more symptoms compared to before treatment. For administration to human patients, the compositions may be formulated by methodologies known to those skilled in the art. The amount of antibody required to bring about therapeutic treatment of a snake bite is not inherently fixed. The amount of antibody administered may vary depending on the extent of the disease and the size of the humans suffering from snake bites. In one case, treatment reduces the infection rate within a population of subjects. Treatment may also result in shorter recovery time, fewer symptoms, or less severe symptoms, or a combination thereof, compared to an untreated subject with a snake bite.

[0214] "Administering" is defined herein as providing one or more compositions to a patient such that the compositions are consequently present in the patient's body. Such administration may be by any route, including, but not limited to, local, local or systemic, subcutaneous, intradermal, intravenous, intra-arterial, intraperitoneal or intramuscular administration (e.g., injection). In one instance, administration is by intradermal injection. In another instance, administration is by subcutaneous injection. In one embodiment, the subject is administered one or more times one of the antibodies or antigen-binding fragments specified herein. In another embodiment, the subject is administered one or more times two of the antibodies or antigen-binding fragments specified herein. In another embodiment, the subject is administered one or more times three of the antibodies or antigen-binding fragments specified herein. In another embodiment, the subject is administered one or more times four of the antibodies or antigen-binding fragments specified herein. Administration may be via any available route, including but not limited to enteral, gastrointestinal, oral, transdermal, intracutaneous, intradermal, subcutaneous, nasal, intravenous, intraperitoneal, intra-arterial, intramuscular, intraosseous, mucosal, aerated, or sublingual administration.

[0215] The actual dosage level of the antibody can be varied to obtain an amount of the active ingredient that is effective in achieving the desired therapeutic response without becoming toxic to the patient. The selected dosage level will depend on a variety of factors, including the activity of the particular antibody used, the route of administration, the number of doses, the excretion rate of the particular antibody to be used, the duration of treatment, other drugs, compounds and / or materials used in combination with the particular composition to be used, the age, sex, weight, condition, overall health and medical history of the patient to be treated, and similar factors.

[0216] The antibodies or antigen-binding fragments described herein can be administered at various dosages over various time frames.

[0217] A physician or veterinarian can easily determine and prescribe the required effective dose (ED50) of the antibody. For example, a physician or veterinarian may start with a dose of the antibody used in the composition at a level lower than what is needed to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. Alternatively, the dose may remain constant.

[0218] The antibody can be administered to the patient via any convenient route, such as those described above. Regardless of the chosen route of administration, the antibody of this disclosure, which can be used in a preferred hydrated form and / or composition, can be formulated into an acceptable dosage form.

[0219] The toxicity and therapeutic efficacy of a compound are, for example, determined by the LD50 (lethal dose for 50% of the population) and ED. 50The effective dose (for treatment in 50% of the population) can be determined by standard procedures in cell cultures or experimental animals. The dose-to-toxicity ratio is the therapeutic index, which can be expressed as the LD50 / ED50 ratio. Compounds that exhibit toxic side effects can be used, but care must be taken to design a delivery system that targets such compounds to the affected tissue in order to minimize potential damage to healthy cells and thereby reduce side effects.

[0220] Data obtained from cell culture assays and / or animal studies can be used to formulate dosage ranges for human use. Doses of such compounds preferably lie within a circulating concentration range containing an ED50 with little to no toxicity. Doses may vary within this range depending on the dosage form and route of administration used. For any compound, the therapeutically effective dose can first be estimated from a cell culture assay. The dose to achieve the circulating plasma concentration containing the IC50 (i.e., the concentration of the test compound that achieves semi-maximal inhibition) determined in cell culture can then be formulated in an animal model. Plasma levels can be measured, for example, by high-performance liquid chromatography. Using such information, a more accurate determination of a useful dose in humans can be made.

[0221] It will be understood that one or more administrations of antibodies or antigen-binding fragments as described herein may be supplemented with one or more additional therapies or drugs, such as nonsteroidal anti-inflammatory drugs (NSAIDs). The most well-known NSAIDs are aspirin, acetaminophen, ibuprofen, and naproxen, all of which are available over-the-counter (OTC) in most countries. Additional, non-exclusive, examples of NSAIDs include, but are not limited to, diflunisal, dex ibuprofen, fenoprofen, ketoprofen, dex ketoprofen, flurbiprofen, oxaprozin, loxoprofen, indomethacin, tolmetin, sulindac, etodolac, ketorolac, diclofenac, aceclofenac, bromfenac, nabumetone, piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, naproxen sodium, acetylsalicylic acid, mefenamic acid, celecoxib, and phenylbutazone. One or more additional therapies or drugs may include, for example, a PLA2 inhibitor. PLA2 inhibitors include, but are not limited to, valesprazib, methylvalesprazib, or a combination thereof.

[0222] Exemplary definition When used herein, the term "about" generally refers to a range (±) that is 2%, 5%, 10%, or 15% greater or less than a number stated in the context of a particular usage. For example, "about 10" would include the range from 8.5 to 11.5. When used herein, the terms "about" and "approximately" are used to modify a number or range of numbers, indicating that deviations of up to about 0.2%, about 0.5%, about 1%, about 2%, about 5%, about 7.5%, or about 10% (or any integer between about 1% and 10%) above and below the value or range remain within the intended meaning of the listed value or range.

[0223] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include multiple subjects unless explicitly indicated otherwise by the context. Thus, for example, a reference to “a method” includes one or more methods and / or steps of the type described herein and / or of the type that would become apparent to those skilled in the art by reading this disclosure.

[0224] An antigen can be a molecule that can be recognized by the immune system, such as a snake toxin. For example, an antigen can be a foreign substance that can induce an immune response. In some cases, the induced immune response may involve the production of antibodies. In some cases, antibodies can recognize, attach to, or bind to the antigen.

[0225] Where used herein, "snake" refers to any venomous snake, including but not limited to: Boiga irregularis (southern giant snake), Boiga cyanea (green giant snake), Boiga dendrophila (mangrove snake), Dispholidus typus (boomslang), Salvadora grahamiae (mountain patchnose snake), Spalerosophis diadema (crowned snake), Tantilla nigriceps (plains blackhead snake), Thelotornis capensis (southern twig snake), Thelotornis kirtlandii (northern twig snake), Toxicodryas blandingii (branding tree snake), Trimorphodon lambda (Sonoran lyre snake), Amphiesma stolatum (buff kaleback), Natrix Tessellate (dice snake), Rhabdophis subminiatus (red-necked keelback), Rhabdophis tigrinus (Japanese keelback), Thamnophis elegans (Western terrestrial garter snake), Thamnophis sirtalis (common garter snake), Ahaetulla nasuta (long-nosed whip snake), Atractaspis bibronii (bibronze's burrowing asp), Atractaspis dahomeyensis (Dahomey's burrowing asp), Atractaspis engaddensis (Palestinian mole snake), Atractaspis microlepidota (small-scaled burrowing asp), Malpolon monspessulanus (Montpellier's snake), Acanthophis antarcticus (common death adder), Aipysurus laevis (olive brown sea snake), Aipysurus duboisii (Dubois sea snake), Austrelaps superbus (lowland pit viper), Cryptophis nigrescens (small ice snake), Demansiaolivacea (Olive Whip Snake), Emydocephalus annulatus (Spotted Sea Snake), Furina tristis (Stevens' Striped Snake), Hydrophis melanocephalus (Black-headed Sea Snake), Hydrophis curtus (Spiny Sea Snake), Hydrophis gracilis (Slender Sea Snake), Hydrophis elegans (Elegant Sea Snake), Hydrophis jerdonii (Corn-noseed Sea Snake), Hydrophis klossi (Slangor Sea Snake), Hydrophis peronii (Horned Sea Snake), Hydrophis belcheri (Belcher's Sea Snake), Hydrophis stricticollis (Bengal Sea Snake), Hydrophis major (Olive-headed Sea Snake), Hydrophis stokesii (large-headed sea snake), Hydrophis melanosoma (black-banded robust sea snake), Hydrophis hardwickii (dorsal-ventral sea snake), Hydrophis cyanocinctus (spotted sea snake), Hydrophis spiralis (narrow-banded sea snake), Hydrophis nigrocinctus (black-banded sea snake), Hydrophis platurus (black-backed sea snake), Hydrophis ornatus (black-spotted sea snake), Hydrophis viperinus (venomous sea snake), Hydrophis schistosus (wart sea snake), Notechis scutatus (mainland tiger snake), Oxyuranus scutellatus (coastal taipan), Oxyuranus temporalis (central range taipan), Pseudechis australis (Margas snake), Pseudechis butleri (Butler's black snake), Pseudechis colletti (Colette's black snake), Pseudechis guttatus (Blue-bellied black snake), Pseudechis papuanus (Papua black snake), PseudechisPorphyriacus (Red-bellied Black Snake), Pseudonaja affinis (Dugait's Snake), Pseudonaja guttata (Speckled Brown Snake), Pseudonaja inframacula (Peninsula Brown Snake), Pseudonaja nuchalis (Western Brown Snake), Pseudonaja textilis (Eastern Brown Snake), Tropidechis carinatus (Clarence River Snake), Aspidelaps lubricus (Cape Coral Snake), Aspidelaps scutatus (Shield-nose Snake), Bungarus fasciatus (Round-tailed Krait), Bungarus caeruleus (Indoor Krait), Bungarus candidus (Blue Krait), Bungarus flaviceps (Red-headed Crate), Bungarus multicinctus (Krait), Dendroaspis viridis (Western Green Mamba), Dendroaspis angusticeps (Eastern Green Mamba), Dendroaspis jamesoni (Jameson's Mamba), Dendroaspis polylepis (Black Mamba), Elapsoidea sundevallii (Sandebaru African Garter Snake), Hemachatus haemachatus (Lincalus), Laticauda colubrina (Blue-spotted Sea Snake), Laticauda laticaudata (Broad-skinned Sea Snake), Laticauda semifasciata (Erabu Sea Snake), Micrurus obscurus (Bolivian Coral Snake), Micrurus frontalis (Southern Coral Snake), Micrurus alleni (Allen's Coral Snake), Micrurus altirostris (Uruguayan Coral Snake), Micrurus clarki (Clark's Coral Snake), Micrurus corallinus (painted coral snake), Micrurus distans (Western Mexican coral snake), Micrurus dumerilii (Dumeril's coral snake), MicrurusMicrurus fulvius (Eastern coral snake), Micrurus hemprichii (Hemprich's coral snake), Micrurus ibiboboca (Kaachinga coral snake), Micrurus lemniscatus (South American coral snake), Micrurus mipartitus (Red-tailed coral snake), Micrurus mosquitensis (Costa Rican coral snake), Micrurus multifasciatus (Striped coral snake), Micrurus nigrocinctus (Central American coral snake), Micrurus pyrrhocryptus (Argentine coral snake), Micrurus spixii (Amazonian coral snake), Micrurus surinamensis (Aquatic coral snake), Micrurus tener (Texas coral snake), Micrurus tschudii (Desert coral snake), Naja siamensis (Indochinese spitting cobra), Naja Naja annulata (Ringed Water Cobra), Naja annulifera (Long-nosed Cobra), Naja ashei (Giant Poisonous Cobra), Naja atra (Taiwanese Cobra), Naja christyi (Conglomerate Water Cobra), Naja haje (Egyptian Cobra), Naja kaouthia (Thai Cobra), Naja katiensis (Western African Poisonous Cobra), Naja melanoleuca (Forest Cobra), Naja mossambica (Mozambican Poisonous Cobra), Naja naja (Indian Cobra), Naja nigricollis (Black-necked Poisonous Cobra), Naja nivea (Cape Cobra), Naja nubiae (Nubian Poisonous Cobra), Naja oxiana (Caspian Cobra), Naja pallida (Red Poisonous Cobra), Naja philippinensis (Northern Philippine Cobra), Naja Samarensis (Samara cobra), Naja sputatrix (Indonesian spitting cobra), Naja sumatrana (Sumatran spitting cobra), Ophiophagus hannah (King cobra), Walterinnesia aegyptia (Western desert cobra), Homalopsisbuccata (Linnaeus's water snake), Myrrophis chinensis (Chinese mud snake), Subsessor bocourti (Bocourt's water snake), Azemiops feae (cobra viper), Agkistrodon bilineatus (patterned pit viper), Agkistrodon contortrix (American pit viper), Agkistrodon piscivorus (swamp pit viper), Agkistrodon taylori (castellana), Agkistrodon laticinctus (broadband copperhead), Atropoides picadoi (Picado's jumping pit viper), Bothriechis lateralis (sidestriped palm pit viper), Bothriechis nigroviridis (black spotted palm pit viper), Bothriechis schlegelii (eyelash palm pit viper), Bothrops diporus (chacolance head), Bothrops erythromelas (Curtin Lancehead), Bothrops insularis (Golden Lancehead Viper), Bothrops jararaca (Jararaka), Bothrops neuwiedi (Neuwiedi Lancehead), Bothrops pauloensis (Blackfaced Lancehead), Bothrops asper (Terciopero), Bothrops atrox (Kaikasa), Bothrops ayerbei (Ayerbei Lancehead), Bothrops caribbaeus (Saint Lucia Lancehead), Bothrops jararacussu (Jararacus), Bothrops lanceolatus (Martiny Lancehead), Bothrops leucurus (Whitetail Lancehead), Bothrops moojeni (Brazil Lancehead), Bothrops alternatus (Urutu), Bothrops cotiara (Cotiara), Bothrops fonsecai (Lance Head of Fonseca), Bothrops itapetiningae (Lance Head of São Paulo), Bothropstaeniatus (Speckled Forest Pit Viper), Bothrops mattogrossensis (Matogrosso Lanzen Otter), Calloselasma rhodostoma (Malayan Pit Viper), Cerrophidion godmani (Godman Mountains Pit Viper), Cerrophidion sasai (Costa Rican Mountains Pit Viper), Crotalus viridis (Prairie Rattlesnake) Crotalus atrox (Western diamondback rattlesnake), Crotalus adamanteus (Eastern diamondback rattlesnake), Crotalus basiliscus (Mexican west coast rattlesnake), Crotalus catalinensis (Santa Catalina Island rattlesnake), Crotalus cerastes (Leftback rattlesnake), Crotalus Cerberus (Arizona black rattlesnake), Crotalus durissus (South American rattlesnake), Crotalus enyo (Baja California rattlesnake), Crotalus horridus (Wood rattlesnake), Crotalus lepidus (Mottled rock rattlesnake), Crotalus mitchellii (San Lucan spotted rattlesnake), Crotalus molossus (Northern black-tailed rattlesnake), Crotalus oreganus (North Pacific rattlesnake), Crotalus Pricei (Western Twin-Spotted Rattlesnake), Crotalus pusillus (Tacita Land Rattlesnake), Crotalus ravus (Mexican Pygmy Rattlesnake), Crotalus ruber (Red Diamond Rattlesnake), Crotalus scutulatus (Mojave Rattlesnake), Crotalus simus (Central American Rattlesnake), Crotalus tigris (Tiger Rattlesnake), Crotalus totonacus (Totonacan Rattlesnake), Crotalus tzabcan (Yucatan Neotropical Rattlesnake), Crotalus willardi (Arizona Ridgenose Rattlesnake), Crotalus Pyrrhus (Southwestern Speckled Rattlesnake), Crotalus vegrandis (Uracoan Rattlesnake), Deinagkistrodon acutus (Hyappoda), Gloydius intermedius (intermediate pit viper), Gloydius blomhoffii (pit viper), Gloydius brevicaudus (long-tailed pit viper), Gloydius halys (Siberian pit viper), Gloydius shedaoensis (snake island pit viper), Gloydiusussuriensis (Ussuri pit viper), Hypnale hypnale (Knobby pit viper), Lachesis melanocephala (Blackhead bushmaster), Lachesis muta (Atlantic bushmaster), Ovophis okinavensis (Omehabu), Porthidium nasutum (Tropical rainforest pit viper), Porthidium ophryomegas (Slender pit viper), Protobothrops elegans (Sakishima pit viper), Protobothrops flavoviridis (Okinawa pit viper), Protobothrops mangshanensis (Manshan pit viper), Protobothrops mucrosquamatus (Taiwan pit viper), Protobothrops tokarensis (Tokara pit viper), Sistrurus catenatus (Masasogai), Sistrurus miliarius (Pygmy rattlesnake), Trimeresurus stejnegeri (Taiwanese Green Pit Viper), Trimeresurus albolabris (White-lipped Green Pit Viper), Trimeresurus erythrurus (Burmese Green Pit Viper), Trimeresurus gramineus (Green Pit Viper), Trimeresurus labialis (Nicobar Bamboo Pit Viper), Trimeresurus macrops (Large-eyed Pit Viper), Trimeresurus malabaricus (Malabar Pit Viper), Trimeresurus popeiorum (Pope Green Pit Viper), Trimeresurus purpureomaculatus (Mangrove Pit Viper), Trimeresurus sumatranus (Sumatran Pit Viper), Tropidolaemus wagleri (Wagler Pit Viper), Metlapilcoatlus mexicanus (Mexican Jumping Pit Viper), Metlapilcoatlus nummifer (Central American Jumping Pit Viper), Atheris squamigera (lizard bush viper), Bitis arietans (puff adder), Bitis Atropos (Cape mountain adder), Bitiscaudalis (horned puff adder), Bitis cornuta (western branched horned adder), Bitis gabonica (Central African Gabon viper), Bitis nasicornis (rhinoceros adder), Bitis parviocula (Ethiopian adder), Bitis rhinoceros (West African Gabon viper), Causus rhombeatus (common night adder), Cerastes cerastes (horned viper), Cerastes gasperettii (Arabian horned viper), Cerastes vipera (Saharan sand viper), Daboia palaestinae (Palestinian viper), Daboia russelii (Russell's viper), Daboia siamensis (Eastern Russell's viper), Daboia mauritanica (Moorish viper), Echis ocellatus (West African carpet viper), Echis carinatus (sawtooth viper), Echis coloratus (painted sawtooth viper), Echis pyramidum (Egyptian sawtooth viper), Macrovipera schweizeri (Milos viper), Macrovipera lebetinus (Levant viper), Montivipera bornmuelleri (Lebanese mountain viper), Montivipera latifii (Latific viper), Montivipera raddei (Armenian mountain viper), Montivipera xanthine (Ottoman viper), Pseudocerastes fieldi (Field horned viper), Pseudocerastes persicus (Persian horned viper), Vipera ammodytes (European horned viper), Vipera aspis (Asp viper), Vipera berus (European viper), Vipera latastei (Latast's viper), Vipera lotievi (Caucasian wild viper), Vipera seoanei (Basque viper), Vipera ursinii (Wild viper), Diadophispunctatus (collared snake), Hydrodynastes gigas (water cobra), Hypsiglena torquata (night snake), Hypsiglena jani (San Luis Stoposi night snake), Leptodeira ashmeadii, or Philodryas olfersii (Liechtenstein green racer).

[0226] Toxins (e.g., snake venom) may be components of modified saliva that contain animal toxins that can aid in the immobilization and / or digestion of prey or defense against dangerous entities. Toxins may be introduced into a target, for example, by biting or spitting.

[0227] The toxin may include a protein or polypeptide. In some embodiments, the toxin may include a mixture of proteins, enzymes, or other substances that are toxic or lethal and may affect the subject. The effects of the toxin on the subject may include effects on biological functions such as blood clotting, blood pressure regulation, or the transmission of nerve or muscle impulses.

[0228] Proteins can constitute 90–95% of the dry weight of a toxin and may be responsible for its biological effects. Toxins may include toxins (e.g., neurotoxins), non-toxic proteins (which may also have pharmacological properties), and / or enzymes, particularly hydrolyzable ones. Enzymes can constitute 80–90% of viper venom and 25–70% of cobra venom and may include digestive hydrolases, L-amino acid oxidases, phospholipases, thrombin-like procoagulants, and kallikrein-like serine proteases or metalloproteinases (hemorrhagic toxins), which can damage vascular endothelium. Polypeptide toxins may include cytotoxins, cardiotoxins, and postsynaptic neurotoxins (e.g., α-bungarotoxin and α-cobratoxin), which can bind to acetylcholine receptors, for example, at neuromuscular junctions. Compounds with low molecular weight (e.g., up to 1.5 kDa) include metals, peptides, lipids, nucleosides, carbohydrates, amines, or oligopeptides that inhibit angiotensin-converting enzyme (ACE) or enhance bradykinin (BPP). Interspecies and intraspecies differences in toxin chemical composition may be geographical and / or ontogenetic. Phosphodiesterases may interfere with the cardiac system, for example, by lowering blood pressure. Phospholipase A2 may cause hemolysis by lysing the phospholipid cell membrane of red blood cells. Amino acid oxidases and proteases may be used in digestion. Amino acid oxidases may induce other enzymes and may be responsible for the yellow color of toxins in some species. Hyaluronidases may increase tissue permeability, accelerating the absorption of other enzymes into tissues. Some snake venoms may contain fasciculins, e.g., mamba (Dendroaspis), which can inhibit cholinesterase, leading to loss of muscle control in prey. Examples of snake venom enzymes are provided in Table 19. [Table 19-1] [Table 19-2]

[0229] Toxins, such as those found in snake venom, can differ in their function. Classes of toxins that may be found in snake venom may include neurotoxins, hemotoxins, cytotoxins, myotoxins, dendrotoxins, cardiotoxins, fasciculins, neutrotoxins, saraphotoxins, and hemorrhagic toxins. A non-limiting list of toxin types found in snake venom and other toxins can be found in Table 20. [Table 20]

[0230] antibody As used herein, the term “antibody” refers to immunoglobulin (Ig), which refers to a polypeptide or protein having an antigen-binding domain or a binding domain homologous thereto. This term further includes “antigen-binding fragment,” and other substitutable terms for similar binding fragments, such as those described below. Native antibodies and native immunoglobulins (Ig) are typically heterotetrameric glycoproteins of about 150,000 daltons, consisting of two identical light chains and two identical heavy chains. Each light chain is typically linked to a heavy chain by one covalent disulfide bond, although the number of disulfide linkages differs between heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has interchain disulfide bridges arranged at regular intervals. Each heavy chain has a variable domain ("V") at one end. H It has ) followed by several constant domains ("C H Each light chain has a variable domain ("V") at one end. L "), and a constant domain ("C") at the other end. L The constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the variable domain of the light chain is aligned with the variable domain of the heavy chain. Certain amino acid residues are thought to form the boundary between the variable domain of the light chain and the variable domain of the heavy chain.

[0231] In some cases, the antibody or its antigen-binding fragment includes an isolated antibody or its antigen-binding fragment, a purified antibody or its antigen-binding fragment, a recombinant antibody or its antigen-binding fragment, a modified antibody or its antigen-binding fragment, or a synthetic antibody or its antigen-binding fragment. It will be understood that the antibodies described herein may be modified as described below or as known in the art.

[0232] Antibodies and antigen-binding fragments described herein can be produced partially or completely by synthesis. Antibodies or antigen-binding fragments may be polypeptides or proteins having a binding domain, which may be an antigen-binding domain or homologous thereto. In one example, an antibody or its antigen-binding fragment can be produced in a suitable in vivo animal model, and then isolated and / or purified.

[0233] Depending on the amino acid sequence of the constant domain of its heavy chain, immunoglobulins (Ig) can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The subunit structures and three-dimensional arrangements of the different classes of immunoglobulins are well known in the art. Ig or a portion thereof may, in some cases, be human Ig. In some cases, C H 3. The domains are derived from immunoglobulins. In some cases, the chain or part of the antibody or its antigen-binding fragment, modified antibody or its antigen-binding fragment, or binder may be derived from Ig. In such cases, Ig may be IgG, IgA, IgD, IgE, or IgM. If Ig is IgG, it may be a subtype of IgG, and the subtypes of IgG may include IgG1, IgG2a, IgG2b, IgG3, and IgG4. In some cases, C HThe three domains may be derived from immunoglobulins selected from the group consisting of IgG, IgA, IgD, IgE, and IgM.

[0234] The "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two distinct types, called kappa ("κ" or "K") or lambda ("λ"), based on the amino acid sequence of their constant domains.

[0235] In this disclosure, where necessary, the following abbreviations (in parentheses) are used in accordance with practice: heavy chain (H chain), light chain (L chain), heavy chain variable region (VH), light chain variable region (VL), complementarity-determining region (CDR), first complementarity-determining region (CDR1), second complementarity-determining region (CDR2), third complementarity-determining region (CDR3), first heavy chain complementarity-determining region (VH CDR1), second heavy chain complementarity-determining region (VH CDR2), third heavy chain complementarity-determining region (VH CDR3), first light chain complementarity-determining region (VL CDR1), second light chain complementarity-determining region (VL CDR2), third light chain complementarity-determining region (VL CDR3), the first heavy chain framework region (FW-H1), the second heavy chain framework region (FW-H2), the third heavy chain framework region (FW-H3), the fourth heavy chain framework region (FW-H4), the first light chain framework region (FW-L1), the second light chain framework region (FW-L2), the third light chain framework region (FW-L3), and the fourth light chain framework region (FW-L4).

[0236] The "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either individually or in combination. Each variable region of the heavy and light chains consists of four framework regions (FWs) connected by three CDRs, also known as hypervariable regions. The CDRs within each chain are held together with CDRs from other chains, in close proximity by the FWs, and contribute to the formation of the antibody's antigen-binding site.

[0237] In relation to antibodies, the term "variable domain" refers to the variable domain of an antibody used for the binding and specificity of each particular antibody to its specific antigen. However, variability is not evenly distributed throughout the antibody's variable domain. Rather, it is concentrated in three segments called hypervariable regions (also known as CDRs) in both the light chain and heavy chain variable domains. The more highly conserved portion of the variable domain is called the "framework region" or "FW". Each of the unmodified heavy and light chain variable domains contains four FRs (FW1, FW2, FW3, and FW4), primarily adopting a β-sheet configuration in which three CDRs are scattered, forming a connected loop and, in some cases, part of a β-sheet structure. The CDRs in each chain are held together with CDRs from other chains, very close together by the FWs, and contribute to the formation of the antibody's antigen-binding site.

[0238] The terms “hypervariable region” and “CDR” as used herein refer to amino acid residues of an antibody involved in antigen binding. A CDR comprises amino acid residues from three sequence regions, which bind to the antigen in a complementary manner. H and V L Each of the chains is known as CDR1, CDR2, and CDR3. It is understood that the CDRs of different antibodies may contain inserts and therefore have different amino acid numbering. The CDR sequences of antibodies and their antigen-binding fragments are provided herein below.

[0239] As used herein, “framework region,” “FW,” or “FR” refers to framework amino acid residues that form part of the antigen-binding pocket or groove. In some embodiments, framework residues form loops that are part of the antigen-binding pocket or groove, and amino acid residues within the loops may or may not be in contact with the antigen. The framework region generally includes the region between the CDR. Loop amino acids of the FR can be evaluated and determined by examining the three-dimensional structure of the antibody heavy chain and / or antibody light chain. The three-dimensional structure can be analyzed for amino acid positions exposed to the solvent, because such positions are more likely to form loops and / or cause contact with the antigen within the antibody variable domain. Some positions exposed to the solvent can tolerate diversity in amino acid sequences, while others (e.g., structural positions) generally have less diversity. The three-dimensional structure of the antibody variable domain can be derived from crystal structure or protein modeling.

[0240] The term “Fc region” is used to define the C-terminal region of an immunoglobulin heavy chain. The “Fc region” may be the native sequence Fc region or a variant Fc region. While the boundaries of the Fc region of an immunoglobulin heavy chain vary, the human IgG heavy chain Fc region is typically defined as extending from the amino acid residue at position Cys226 or Pro230 to its carboxyl terminus. The numbering of residues within the Fc region is based on EU index numbering, as seen in Kabat et al., (Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991). The Fc region of an immunoglobulin generally consists of two constant domains, C H 2 and C H Includes 3.

[0241] In one instance, an antibody or antigen-binding fragment may contain a variable region. The variable region may be either a variable region of the antibody light chain or a variable region of the antibody heavy chain, either alone or in combination. Each variable region of the heavy and light chains consists of four framework regions (FRs) connected by three complementarity-determining regions (CDRs), also known as hypervariable regions. The CDRs within each chain are held together with CDRs from other chains, in close proximity via FRs, and contribute to the formation of the antibody's antigen-binding site.

[0242] In one case, an antibody or antigen-binding fragment may include a light chain region, a heavy chain region, or both a light chain and a heavy chain region that confer specific binding, for example, to a toxin. In another case, an antibody or antigen-binding fragment may include a constant region. The constant region may include the constant region of the antibody light chain, either alone or in combination with the constant region of the antibody heavy chain.

[0243] In this disclosure, “antibodies” are useful but are not limited to monoclonal antibodies, polyclonal antibodies, chimeric antibodies, bispecific antibodies, multispecific antibodies, heteroconjugate antibodies, humanized antibodies, human antibodies, deimmunized antibodies, mutants thereof, fusions thereof, immunoconjugates thereof, antigen-binding fragments thereof, and / or any other modified configuration of immunoglobulin molecules containing antigen-recognizing sites of the required specificity, including glycosylated variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies.

[0244] As used herein, “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies constituting this population are identical except for naturally occurring mutations that may be present in small amounts. In contrast to polyclonal antibody preparations, which typically contain different antibodies against different determinants (epitopes), each monoclonal antibody is against a single determinant on an antigen (epitope). The modifier “monoclonal” indicates a characteristic of the antibody that it is obtained from a substantially homogeneous population of antibodies, and this modifier should not be interpreted as requiring the production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with this disclosure can be produced by the hybridoma method first described by Kohler and Milstein, 1975, Nature, 256:495, or by recombinant DNA methods such as those described in U.S. Patent No. 4,816,567. Monoclonal antibodies can also be isolated from phage libraries generated using techniques such as those described in McCafferty et al., 1990, Nature, 348:552-554. Other methods are known in the art and are intended for use herein.

[0245] As used herein, the term "humanized" antibody refers to a form of non-human (e.g., mouse) antibody that is a specific chimeric immunoglobulin, immunoglobulin chain, or fragment thereof containing a minimal sequence derived from a non-human immunoglobulin. In most cases, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's complementarity-determining region (CDR) are replaced by residues from the CDR of a non-human species (donor antibody), such as mouse, rat, or rabbit, having the desired antibody specificity, affinity, and bioactivity. In some cases, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, a humanized antibody may include residues not found in the recipient antibody or the transferred CDR and framework sequence, but which are included to further refine and optimize antibody performance. Generally, a humanized antibody will contain at least one, usually two, substantially all of the variable domains, where all or substantially all of the CDR region corresponds to that of a non-human immunoglobulin, and all or substantially all of the FR region corresponds to that of the human immunoglobulin consensus sequence. Humanized antibodies will also include at least a portion of the constant region or domain (Fc) of an immunoglobulin, typically that of human immunoglobulin. The antibody may have a modified Fc region, for example, as described in WO99 / 58572. Other forms of humanized antibodies have one or more CDRs (one, two, three, four, five, or six) that are modified relative to the original antibody, and these CDRs are also referred to as one or more CDRs "derived" from one or more CDRs from the original antibody.

[0246] If necessary, the antibodies or antigen-binding fragments described herein can be evaluated for immunogenicity and, if necessary, deimmunized (i.e., made less immunoreactive by modifying one or more T cell epitopes). As used herein, “deimmunized antibody” means that one or more T cell epitopes in the antibody sequence have been modified such that the T cell response after administration of the antibody to a subject is reduced compared to an unimmunized antibody. Analysis of the immunogenicity of the antibodies and antigen-binding fragments described herein and the T cell epitopes present therein can be performed using software and specific databases known in the art. Illustrative software and databases include iTope®, developed by Antitope of Cambridge, England. iTope® is an in silico technology for the analysis of peptides that bind to human MHC class II alleles. The iTope® software predicts peptides that bind to human MHC class II alleles, thereby performing an initial screening for the location of such “potential T cell epitopes.” iTope® software predicts favorable interactions between the amino acid side chains of peptides and specific binding pockets within the binding grooves of 34 human MHC class II alleles. The critical binding residue locations are obtained by in silico generation of 9mer peptides with a single amino acid duplicate across the variable region sequence of the test antibody. Each 9mer peptide can be tested against each of the 34 MHC class II allotypes and scored based on their potential "fit" and interaction with the MHC class II binding groove. Peptides that produce a high average binding score (>0.55 in iTope® scoring function) for >50% of the MHC class II alleles are considered potential T cell epitopes.In such regions, the core 9 amino acid sequence for peptide bonding within the MHC class II groove is analyzed to determine MHC class II pocket residues (P1, P4, P6, P7, and P9) and potential T cell receptor (TCR) contact residues (P-1, P2, P3, P5, and P8). After identifying any T cell epitopes, amino acid residue changes, substitutions, additions, and / or deletions can be introduced to remove the identified T cell epitope. Such changes can be made to preserve antibody structure and function but still remove the identified epitope. Exemplary changes may include, but are not limited to, conservative amino acid changes.

[0247] Antibodies may be human antibodies. As used herein, “human antibody” means an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human, and / or produced using any of the techniques for producing human antibodies known in the art or disclosed herein. This definition of a human antibody includes antibodies comprising at least one human heavy chain polypeptide or at least one human light chain polypeptide. One such example is an antibody comprising a mouse light chain and a human heavy chain polypeptide. Human antibodies can be produced using a variety of techniques known in the art. In one embodiment, human antibodies are selected from a phage library, which expresses human antibodies (Vaughan et al., 1996, Nature Biotechnology, 14:309-314; Sheets et al., 1998, PNAS USA, 95:6157-6162; Hoogenboom and Winter, 1991, J. Mol. Biol., 227:381; Marks et al., 1991, J. Mol. Biol., 222:581). Human antibodies can also be produced by introducing human immunoglobulin loci into transgenic animals, such as mice, in which endogenous immunoglobulin genes are partially or completely inactivated. This approach is described in U.S. Patents No. 5,545,807, 5,545,806, 5,569,825, 5,625,126, 5,633,425, and 5,661,016. Alternatively, human antibodies can be prepared by immortalizing human B lymphocytes that produce antibodies against a target antigen (such B lymphocytes may be recovered from a subject or immunized in vitro). See, for example, Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., 1991, J. Immunol., 147 (1):86-95; and U.S. Patent No. 5,750,373.

[0248] Bispecific antibodies are antibodies that have binding specificity to at least two different antigens and can be prepared using the antibodies disclosed herein. Methods for producing bispecific antibodies are known in the art (see, for example, Suresh et al., 1986, Methods in Enzymology 121:210). Traditionally, recombinant production of bispecific antibodies has been based on the co-expression of two immunoglobulin heavy-light chain pairs using two heavy chains with different specificities (Millstein and Cuello, 1983, Nature, 305, 537-539). Bispecific antibodies may consist of a hybrid immunoglobulin heavy chain having a first binding specificity in one arm and a hybrid immunoglobulin heavy-light chain pair (providing a second binding specificity) in the other arm. This asymmetric structure, in which only half of the bispecific molecule has an immunoglobulin light chain, facilitates the separation of desired bispecific compounds from undesirable immunoglobulin chain combinations. This approach is described in PCT Publication No. WO94 / 04690.

[0249] One approach to producing bispecific antibodies involves fusing an antibody variable domain having the desired binding specificity (antibody-antigen synthesis site) to an immunoglobulin constant domain sequence. Preferably, the fusion is with an immunoglobulin heavy chain constant domain that includes at least a portion of the hinge, CH2, and CH3 regions. It is preferable that at least one of the fusions has a first heavy chain constant region (CH1) containing the site required for light chain binding. The immunoglobulin heavy chain fusion and, if desired, the DNA encoding the immunoglobulin light chain are inserted into separate expression vectors and co-transfected into a suitable host organism. This provides great flexibility in adjusting the relative proportions of the three polypeptide fragments when a non-uniform ratio of the three polypeptide chains used in construction yields the optimal yield. However, when the expression of at least two polypeptide chains in the same ratio results in a high yield, or when the ratio is not particularly significant, it is possible to insert the coding sequences of two or all three polypeptide chains into a single expression vector.

[0250] Heteroconjugate antibodies, which contain two antibodies covalently linked together, are also within the scope of this disclosure. Such antibodies have been used to target immune system cells to undesirable cells (U.S. Patent No. 4,676,980). Heteroconjugate antibodies can also be prepared using any convenient crosslinking method. Suitable crosslinking agents and techniques are known in the art and are described, for example, in U.S. Patent No. 4,676,980.

[0251] The “chimeric” form of non-human (e.g., mouse) antibodies includes chimeric antibodies that contain a minimal sequence derived from non-human Ig. In most cases, chimeric antibodies are typically mouse antibodies in which at least a portion of the immunoglobulin constant region (Fc) of a human immunoglobulin is inserted in place of the mouse Fc.

[0252] Chimeric or hybrid antibodies can also be prepared in vitro using known methods of synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using disulfide exchange reactions or by forming thioether bonds. Examples of suitable reagents for this purpose include iminothiolates and methyl-4-mercaptobutylimidate.

[0253] Antibodies and their antigen-binding fragments, modified antibodies and their antigen-binding fragments, and binders are provided herein, which specifically bind to one or more epitopes on one or more target antigens. In one case, the binder specifically binds to an epitope on a single antigen. In another case, the binder is bivalent and specifically binds to two distinct epitopes on a single antigen, or to two distinct epitopes on two distinct antigens. In yet another case, the binder is polyvalent (i.e., trivalent, tetravalent, etc.) and binds to three or more distinct epitopes on a single antigen, or to three or more distinct epitopes on two or more (multiple) antigens.

[0254] Functional fragments of any of the antibodies described herein are also intended. The terms “antigen-binding moiety,” “antigen-binding fragment,” “antigen-binding domain,” “antibody fragment,” or “functional fragment of an antibody” are used herein synonymously to refer to one or more fragments of an antibody that possess the ability to specifically bind to an antigen. Representative antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, scFv, dsFv, variable heavy chain domain, variable light chain domain, variable NAR domain, bispecific scFv, bispecific Fab2, tripspecific Fab3, AVIMER®, minibody, diabody, maxibody, camelid, VHH, intrabody, fusion protein containing an antibody moiety (e.g., domain antibody), single-chain bound polypeptide, scFv-Fc, or Fab-Fc.

[0255] The "F(ab')2" and "Fab" portions can be produced by treating Ig with proteases such as pepsin and papain, and include antigen-binding moieties generated by digesting immunoglobulin near the disulfide bond present between the hinge regions in each of the two heavy chains. For example, papain cleaves IgG upstream of the disulfide bond present between the hinge regions in each of the two heavy chains, V L and C L The light chain and V are composed of (the light chain constant region). H and C Hγ1 Pepsin generates two homologous antibody fragments, each consisting of a heavy chain fragment (the γ1 region in the constant region of the heavy chain) and connected by a disulfide bond at their C-terminal regions. Each of these two homologous antibody fragments is called Fab'. Pepsin also cleaves IgG downstream of the disulfide bond between the hinge regions in each of the two heavy chains, generating an antibody fragment slightly larger than the two aforementioned Fab' fragments connected at the hinge region. This antibody fragment is called F(ab')2.

[0256] The Fab fragment consists of a constant domain of the light chain and a first constant domain (C) of the heavy chain. H 1) also contains. The Fab' fragment is a heavy chain C containing one or more cysteines from the antibody hinge region. H It differs from the Fab fragment in the addition of a few residues at the carboxyl terminus of one domain. Fab'-SH is the herein designation for Fab' in which the cysteine ​​residue of the constant domain has a free thiol group. The F(ab')2 antibody fragment was originally produced as a pair of Fab' fragments having a hinged cysteine ​​between them. Other chemical couplings of antibody fragments are also known.

[0257] As used herein, "Fv" refers to an antibody fragment containing a complete antigen recognition and antigen-binding site. This region consists of a dimer of one heavy chain and one light chain variable domain closely associated non-covalently or covalently (disulfide-linked Fv is described in the Art, Reiter et al. (1996) Nature Biotechnology 14:1239-1245). In this structure, the three CDRs of each variable domain interact to form V H -V L Define the antigen-binding site on the surface of the dimer. In summary, V H and V L One or more combinations of CDRs from each of the chains confer antigen-binding specificity to the antibody. For example, it can be understood that, for instance, CDRH3 and CDRL3 confer antigen-binding specificity to the recipient antibody or its antigen-binding fragment. H and V L It may be sufficient to confer antigen-binding specificity to the antibody when transferred to the chain, and this combination of CDRs can be tested for binding, specificity, affinity, etc., using any of the techniques described herein. Even a single variable domain (or half of the Fv containing only the three antigen-specific CDRs) has the ability to recognize and bind to the antigen, although it is likely to have lower specificity or affinity than when combined with a second variable domain. Furthermore, the two domains of the Fv fragment (V L and V H Although these are encoded by separate genes, they can be joined using a recombination method with a synthetic linker, and this recombination method makes it possible to create them as a single protein chain, in which case V L Region and V HThe regions pair up to form a monovalent molecule (also known as a single-chain Fv (scFv); Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; and Osbourn et al. (1998) Nat. Biotechnol. 16:778). Such scFvs are also intended to be included within the scope of the term "antigen-binding moiety" of an antibody. Any V of a particular scFv H and V L The sequence can be ligated to an Fc region cDNA or genomic sequence to generate an expression vector encoding a complete Ig (e.g., IgG) molecule or other isotype. V can be used to generate Fab, Fv, or other fragments of Ig using either protein chemistry or recombinant DNA technology. H and V L You can also use this.

[0258] "Single-chain Fv" or "sFv" antibody fragments are antibodies of V H and V L It contains domains, and these domains are present in a single polypeptide chain. In some embodiments, the Fv polypeptide contains a polypeptide linker that allows the sFv to form a structure desirable for antigen binding. H Domain and V L Further inclusions are included between domains. For an overview of sFv, see, for example, Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269–315 (1994).

[0259] The term "AVIMER®" refers to a class of human-derived therapeutic proteins independent of antibodies and antibody fragments, comprising several modules called A domains (also known as class A modules, complement-type repeats, or LDL-receptor class A domains) and a reusable binding domain. They were developed from human extracellular receptor domains by in vitro exon shuffling and phage display (Silverman et al., 2005, Nat. Biotechnol. 23:1493-1494; Silverman et al., 2006, Nat. Biotechnol. 24:220). The resulting proteins may contain multiple independent binding domains that can exhibit improved affinity and / or specificity compared to single epitope-binding proteins. Each of the 217 known human A-domains contains approximately 35 amino acids (approximately 4 kDa), and these domains are separated by linkers that average 5 amino acids in length. The native A-domain rapidly and efficiently folds into a uniform, stable structure, primarily mediated by calcium binding and disulfide formation. Only 12 conserved amino acid scaffolding motifs are required for this common structure. The final result is a single protein chain containing multiple domains, each exhibiting a distinct function. Each domain of the protein binds independently, and the energetic contribution of each domain is additive.

[0260] Antigen-binding polypeptides also include heavy chain dimers, such as antibodies from camelids and sharks. Camelid and shark antibodies contain homodimer pairs of two chains (neither of which has a light chain) consisting of a V-like domain and a C-like domain. H Since the region does not need to form hydrophobic interactions with the light chain, the region within the heavy chain that normally comes into contact with the light chain is transformed into a hydrophobic amino acid residue in camelids. H The domain is V HHThis is called a domain. Shark Ig-NAR contains a homodimer of one variable domain (called the V-NAR domain) and five C-like constant domains (C-NAR domains). In camelids, the diversity of the antibody repertoire is V H or V HH Determined by CDR1, 2, and 3 within the region. Camel V HH The CDR3 region within this area is characterized by its relatively long length, averaging 16 amino acids (Muyldermans et al., 1994, Protein Engineering, 7(9): 1129). This is significantly different from the CDR3 region of many other species of antibodies. For example, mouse V H CDR3 has an average of 9 amino acids. A library of camelid-derived antibody variable regions that maintains in vivo diversity of camelid variable regions can be prepared, for example, by the method disclosed in U.S. Patent Application No. 20050037421.

[0261] As used herein, “maxibody” refers to a divalent scFv covalently attached to the Fc region of an immunoglobulin. See, for example, Fredericks et al., Protein Engineering, Design & Selection, 17:95-106 (2004) and Powers et al., Journal of Immunological Methods, 251:123-135 (2001).

[0262] As used herein, "dsFv" may refer to an Fv fragment obtained by introducing Cys residues to suitable sites in the heavy chain variable region and the light chain variable region, respectively, and then stabilizing the heavy chain variable region and the light chain variable region by disulfide bonds. The sites within each chain to which Cys residues should be introduced can be determined based on a higher-order structure predicted by molecular modeling. In this disclosure, for example, the higher-order structure is predicted from the amino acid sequences of the heavy chain variable region and the light chain variable region of the antibody, and then DNA encoding each of the heavy chain variable region and the light chain variable region into which mutations have been introduced based on such predictions is constructed. The DNA constructs are then introduced into a suitable vector and prepared from transformants obtained by transformation with the aforementioned vector.

[0263] Antibody single-chain variable region fragments ("scFv") are described herein. Single-chain variable region fragments can be prepared by ligating light and / or heavy chain variable regions using short-chain linked peptides. Bird et al. (1988) Science 242:423-426. Single-chain variants can be produced by recombination or synthetically. Automated synthesizers can be used for the synthetic production of scFv. For the recombinant production of scFv, a suitable plasmid containing the polynucleotide encoding scFv can be introduced into a suitable host cell, either a eukaryotic cell, e.g., yeast, plant, insect, or mammalian cell, or a prokaryotic cell, e.g., E. coli. The polynucleotide encoding the desired scFv can be prepared by routine manipulation, such as polynucleotide ligation. The resulting scFv can be isolated using standard protein purification techniques known in the art.

[0264] Diabodies can be single-chain antibodies. Diabodies can be bivalent, bispecific antibodies in which the VH and VL domains are expressed on a single polypeptide chain, by pairing a domain with a complementary domain on another chain using a linker that is too short to allow pairing between two domains on the same chain, thereby creating two antigen-binding sites (see, for example, Holliger, P., et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993); and Poljak, RJ, et al., Structure, 2:1121-1123 (1994)).

[0265] As used herein, “mini-body” refers to an scFv fused to CH3 via a peptide linker (hingeless) or via an IgG hinge as described in Olafsen, et al., Protein Eng Des Sel., April 2004; 17(4):315-23.

[0266] As used herein, “intrabody” refers to a single-chain antibody capable of demonstrating intracellular expression and manipulating intracellular protein function (Biocca, et al., EMBO J. 9:101-108, 1990; Colby et al., Proc Natl Acad. Sci. USA. 101:17616-21, 2004). Intrabodies containing a cellular signaling sequence that holds an antibody construct within an intracellular region can be produced as described by Mhashilkar et al., (EMBO J., 14:1542-51, 1995) and Wheeler et al. (FASEB J. 17:1733-5. 2003). A transbody is a cell-permeable antibody in which the transduction domain (PTD) of a protein is fused with a single-chain variable fragment (scFv) antibody (Heng et al., Med Hypotheses. 64:1105-8, 2005).

[0267] The binder can be polymerized using suitable linkers known in the art. A non-limiting example of a linked peptide is (GGGGS)3 (SEQ ID NO: 283), which bridges approximately 3.5 nm between the carboxyl terminus of one variable region and the amino terminus of the other variable region. Linkers of other sequences have been designed and used. Bird et al. (ibid.). The linkers can be further modified for additional functions such as drug adhesion or adhesion to a solid support.

[0268] An "epitope" refers to the portion of an antigen or other macromolecule that can form a binding interaction with the variable region binding pocket of an antibody. Such a binding interaction may manifest as an intermolecular contact with one or more amino acid residues of one or more CDRs. Antigen binding may occur, for example, with CDR3 or a pair of CDR3s, or in some cases, with V H and V L The interaction may include all six CDRs in the chain. Epitopes may be linear peptide sequences ("continuous") or composed of non-adjacent amino acid sequences ("contiguous" or "discontinuous"). Antibodies can recognize one or more amino acid sequences, and therefore, an epitope can define more than one distinct amino acid sequence. Epitopes recognized by antibodies can be determined by peptide mapping and sequence analysis techniques well known to those skilled in the art. Binding interactions manifest as intermolecular contact between the epitope on the antigen and one or more amino acid residues of the CDR.

[0269] Antibodies selectively bind to a target if they bind to that target with greater affinity, avidity, ease, and / or for a longer duration than other substances. For example, an antibody or antigen-binding fragment that selectively binds to, for example, a snake venom toxin is an antibody or antigen-binding fragment that binds to this epitope with greater affinity, avidity, ease, and / or for a longer duration than a protein that is not a snake venom toxin.

[0270] The term "kon," as used herein, is intended to refer to the rate constant for the association of an antibody with an antigen.

[0271] When used herein, the term "Koff" is intended to refer to the rate constant for the dissociation of an antibody from an antibody / antigen complex.

[0272] As used herein, the term “avidity” refers to the resistance of a complex of two or more drugs to dissociation after dilution. Apparent affinity can be determined by methods such as enzyme-linked immunosorbent assay (ELISA) or any other technique well known to those skilled in the art. Avidity can be determined by methods such as scatchard analysis or any other technique.

[0273] As used herein, the term "affinity" refers to the equilibrium constant for the reversible binding of two drugs, where binding affinity (K) is the constant for binding affinity. D It is expressed as ). In some cases, K D k can refer to the rate constant for the dissociation of an antibody from an antibody or antigen-binding fragment / antigen complex. off k can refer to the rate constant for the association of an antibody or antigen-binding fragment with an antigen. on Binding affinity can be expressed as a ratio to . Binding affinity can be determined using methods known in the art, including, for example, surface plasmon resonance (SPR; Biacore), Kinexa Biocensor, scintillation proximity assay, enzyme-linked immunosorbent assay (ELISA), ORIGEN immunoassay (IGEN), fluorescence quenching, fluorescence transfer, yeast display, or any combination thereof. Binding affinity can also be screened using a suitable bioassay. Binding affinity (K) of the antibody or antigen-binding fragment described herein D) may be less than: 600nM, 590nM, 580nM, 570nM, 560nM, 550nM, 540nM, 530nM, 520nM, 510nM, 500nM, 490nM, 480nM, 470nM, 460nM, 450nM, 440nM, 430nM, 420nM, 410nM, 400nM, 390nM, 380nM, 370nM, 360nM, 350nM, 340nM, 330nM, 320nM, 310nM, 300nM, 290nM, 280nM, 270nM, 260nM, 250nM, 240nM, 230nM, 220nM, 210 nM, 200nM, 190nM, 180nM, 170nM, 160nM, 150nM, 140nM, 130nM, 120nM, 110nM , 100nM, 90nM, 80nM, 70nM, 50nM, 50nM, 49nM, 48nM, 47nM, 46nM, 45nM, 44nM, 4 3nM, 42nM, 41nM, 40nM, 39nM, 38nM, 37nM, 36nM, 35nM, 34nM, 33nM, 32nM, 31n M, 30nM, 29nM, 28nM, 27nM, 26nM, 25nM, 24nM, 23nM, 22nM, 21nM, 20nM, 19nM, 1 8nM, 17nM, 16nM, 15nM, 14nM, 13nM, 12nM, 11nM, 10nM, 9nM, 8nM, 7nM, 6nM, 5n M, 4nM, 3nM, 2nM, 1nM, 990pM, 980pM, 970pM, 960pM, 950pM, 940pM, 930pM, 920 pM, 910pM, 900pM, 890pM, 880pM, 870pM, 860pM, 850pM, 840pM, 830pM, 820pM , 810pM, 800pM, 790pM, 780pM, 770pM, 760pM, 750pM, 740pM, 730pM, 720pM, 71 0pM, 700pM, 690pM, 680pM, 670pM, 660pM, 650pM, 640pM, 630pM, 620pM, 610p M, 600pM, 590pM, 580pM, 570pM, 560pM, 550pM, 540pM, 530pM, 520pM, 510pM, 5 00pM, 490pM, 480pM, 470pM, 460pM, 450pM, 440pM, 430pM, 420pM, 410pM, 400 pM, 390pM, 380pM, 370pM, 360pM, 350pM, 340pM, 330pM, 320pM, 310pM, 300pM,290pM, 280pM, 270pM, 260pM, 250pM, 240pM, 230pM, 220pM, 210pM, 200pM, 190pM, 180pM, 170pM, or any integer between these.

[0274] The terms “polypeptide,” “oligopeptide,” “peptide,” and “protein” are used synonymously herein and generally refer to polymers of amino acids of any length. Polymers may be linear or branched, may contain modified amino acids, and may have non-amino acid intercalations. These terms also encompass amino acid polymers modified naturally or by intervention, e.g., disulfide bond formation, glycosylation, lipidization, acetylation, phosphorylation, or any other manipulation or modification, e.g., conjugation with a labeling component. This definition also includes, for example, polypeptides containing one or more analogs of amino acids (including, e.g., non-natural amino acids), as well as polypeptides containing other suitable modifications. Because the polypeptides of this disclosure are antibody-based, it is understood that polypeptides may exist as single chains or as associated chains.

[0275] In this specification, the terms “polynucleotide” and “nucleic acid” as used synonymously generally refer to polymers of nucleotides of any length, including DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or analogs thereof, or any substrate that can be incorporated into the polymer by DNA or RNA polymerase. Polynucleotides may include modified nucleotides, such as methylated nucleotides and their analogs. If present, modifications to the nucleotide structure may be conjugated before or after the polymer is assembled. Non-nucleotide components may also be intercalated in the nucleotide sequence. Polynucleotides may be further modified after polymerization, such as by conjugation with labeling components. Other types of modifications include, for example, "caps," substitutions of one or more naturally occurring nucleotides in analogs, internucleotide modifications, such as those in uncharged linkages (e.g., methyl phosphonate, phosphotryester, phosphoramidate, carbamate, etc.) and charged linkages (e.g., phosphorothioate, phosphorodithioate, etc.), pendant portions, such as those containing proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those containing intercalators (e.g., acridine, psoralen, etc.), those containing chelating agents (e.g., metals, radiometals, boron, metal oxides, etc.), those containing alkylating agents, those having modified linkages (e.g., alpha-anomeric nucleic acids, etc.), and the unmodified form of polynucleotides.

[0276] Furthermore, any of the hydroxyl groups originally present in the sugar can be replaced with, for example, a phosphonic acid group or a phosphate group, protected with a standard protecting group, activated to prepare additional linkages to additional nucleotides, or conjugated to a solid support. The 5' and 3' terminal OH groups can be phosphorylated or replaced with amines or organic capping group moieties having 1 to 20 carbon atoms. Other hydroxyls can also be derivatized with standard protecting groups. The polynucleotide may also contain similar forms of ribose or deoxyribose sugars, such as 2'-O-methyl-, 2'-O-allyl, 2'-fluoro- or 2'-azid-ribose, carbocyclic sugar analogs, alpha-anomeric sugars, epimeric sugars, e.g., arabinose, xylose or lyxose, pyranose sugars, furanose sugars, sedoheptulose, acyclic analogs, and debasalized nucleoside analogs, e.g., methylribose. One or more phosphodiester linkages can be replaced with alternative linkage groups. These alternative linking groups include, but are not limited to, embodiments in which the phosphate is replaced with P(O)S ("thioate"), P(S)S ("dithioate"), (O)NR2 ("amidate"), P(O)R, P(O)OR', CO, or CH2 ("formacetal"), where R or R' is independently H or a substituted or unsubstituted alkyl (1-20C) containing, as may be, an ether (--O--) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl, or aralkyl. Not all linkages in the polynucleotide need to be identical. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

[0277] As used herein, “identity” may mean the percentage of identical nucleotide or amino acid residues at corresponding positions in two or more sequences when sequences are aligned (taking into account gaps and insertions) to maximize sequence matching. Identity can be readily calculated by known methods, including but not limited to those described in Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988). The method for determining identity is designed to yield the maximum match between the sequences being tested. Furthermore, the method for determining identity is systematized in a publicly available computer program.Computer programming methods for determining the identity between two sequences include, but are not limited to, the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1): 387 (1984)), BLASTP, BLASTN, and FASTA (Altschul, SF et al., J. Molec. Biol. 215: 403-410 (1990) and Altschul et al. Nuc. Acids Res. 25: 3389-3402 (1997)). The BLAST X program is available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol. Biol. 215: 403-410 (1990)). Identity can also be determined using the well-known Smith-Waterman algorithm.

[0278] The desired sequence identity range is approximately 80% to approximately 100% and integer values ​​in between. Generally, this disclosure includes sequences having approximately 80%, approximately 81%, approximately 82%, approximately 83%, approximately 84%, approximately 85%, approximately 86%, approximately 87%, approximately 88%, approximately 89%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identity with any sequence provided herein. The letters "X" or "Xaa," when used in amino acid sequences herein, are intended to indicate that any of the 20 standard amino acids can be substituted at this position unless otherwise specified. [Examples]

[0279] The Application can be better understood by referring to the following non-limiting embodiments provided as exemplary embodiments of the Application. These embodiments are presented to better illustrate the embodiments, but should not be considered to limit the scope of the Application in any way.

[0280] (Example 1) Dynamics of antitoxin antibodies Dynamic measurements were performed to calculate the binding affinity of two antibodies to recombinant alpha-neurotoxins from mamba, taipan, and cobra snakes. Alpha-neurotoxins were expressed and purified by affinity chromatography. Antibodies isolated from subjects previously bitten by snakes were incubated with alpha-neurotoxins, and the dynamics of alpha-neurotoxin binding to the antibodies were measured. Dynamic data are provided in Tables 21A and 21B. [Table 21A] [Table 21B]

[0281] (Example 2) Toxicity of toxins in mice The toxicity of different doses of neurotoxins in mice was determined. Recombinant alpha-neurotoxins from various snake species were expressed and purified by affinity chromatography. Intraperitoneal doses greater than the LD50 (the dose at which 50 percent of mice die after the compound) of each toxin were administered to mice (5 mice per toxin group). In addition to general observations, the body weight and rectal temperature of each mouse were measured before toxin administration, again every 20 minutes after administration up to 2 hours, and then 24 hours after administration. Mice that recovered or were unaffected by the given dose of toxin showed complete recovery within approximately 2–4 hours after administration and remained healthy beyond 24 hours post-administration. For the given doses described, if the given toxin dose was lethal, the mice died within 60 minutes after administration or were euthanized. Survival data are presented in Table 22: [Table 22]

[0282] (Example 3) Protective effect of antibodies against toxins in mice Individual antibody clones of the antibodies tested in Example 1 were tested for their ability to neutralize the toxin tested in Example 2 by pre-mixing the antibodies with neurotoxin in an Eppendorf tube and incubating at room temperature for 1 hour. The mixture was then administered to a group of mice (n=5) and lethality was observed as described above. Table 23 shows the neutralization of lethal snake neurotoxin by the antibody clones. [Table 23]

[0283] The mice showed no signs of disease. The molar ratio of the toxin to the antibody (considering bivalentity) is shown. (Example 4) Information on snakes, toxins, and sequences [Table 24-1] [Table 24-2] [Table 25-1] [Table 25-2] [Table 25-3] [Table 26-1] [Table 26-2] [Table 26-3] [Table 26-4] [Table 26-5] [Table 26-6] [Table 26-7]

[0284] (Example 5) Example treatment method A randomized, double-blind, controlled trial was conducted to compare the effects of low-dose and high-dose treatments. Patients presenting hematological or neurological evidence of systemic venom injection within 24 hours of a snake bite were included in the study. Patients were randomly assigned to receive either a high-dose or low-dose treatment with a composition comprising one or more antibodies or antigen-binding fragments as described herein.

[0285] The primary endpoint measures, but not limited to, include (1) a composite endpoint: the number of patients who die, require assisted ventilation, or exhibit worsening neurotoxicity as defined by the appearance of two new neurotoxic signs or the appearance of a severe neurotoxic sign (e.g., loss of pharyngeal reflex or paradoxical breathing), and / or the number of patients who have a serious adverse event.

[0286] (Example 6) Exemplary mouse model to establish MTD to demonstrate the effectiveness of the toxin The toxicity / tolerance to toxins from each species was determined in mice by delivering intraperitoneal doses sufficient to achieve a dose greater than the lethal dose 50 (LD50: 50% of mice died after compound administration).

[0287] Body weight and rectal temperature were measured in each mouse before toxin administration. The toxin was administered intraperitoneally to each mouse (5 mice per group). Rectal temperature measurements and general observations were performed every 20 minutes after administration, up to 2 hours, and then 24 hours after administration. For the given toxins tested, mice that recovered or were not affected by the given dose of toxin showed complete recovery 2 to 4 hours after administration and remained healthy beyond 24 hours. If the given dose of toxin was lethal due to a rapid drop in rectal temperature (>2 degrees from baseline) for the given dose, the mouse died within 60 minutes after administration or was euthanized. Table 27 provides a description of the treatment groups. [Table 27]

[0288] (Example 7) In vivo effects of antitoxin antibodies Experiments were conducted to determine the >LD50 of purified recombinant snake alphaneurotoxin when treated with (1) S-neurotoxin + SNEURO_P01_B11, (2) S-neurotoxin + SNEURO_P01_D9, or (3) an antibody-free control (S-neurotoxin). Survival rates are shown as mice that did not have any significant changes in body temperature or whose body temperature returned to normal up to 2 hours and beyond (Table 28). [Table 28]

[0289] (Example 8) Antibody Library This example demonstrates the production of an antibody library from highly immunized subjects. Highly Immunized Subjects (HiS). The inventors collected B-cell samples from human subjects with a 17-year history of immunization to snake venom by self-administering diluted doses of toxins from taipan, cobra, rattlesnake, coral snake, krait, tiger snake, eastern brown snake, and mamba. These subjects gradually increased the toxin concentration and acquired sufficient antibody titers to withstand all snake bites without succumbing to death or amputation. These HiS provide evidence of administration of over 700 escalating doses of toxin and over 200 raw bites.

[0290] Antitoxin antibody library construction. In the passive research plan, 20 ml of peripheral blood was collected on day 0 and 28 days prior to the predetermined immunization schedule used by highly immunized subjects, which had been established over a 17-year period to maintain their high level of immunity. PBMCs were isolated by Ficol separation, and cDNA was extracted. Antibody variable domain (VH), Vκ, and Vλ diversity was extracted using a primer set designed for simultaneous amplification of the human repertoire for both high-throughput sequencing and antibody presentation library construction. Barcoded library samples were sequenced using MiSeq 2×300 paired-end reads to a depth of 500,000 reads per library. The same material was digested and cloned into scFv-pIII fusion presentation vectors, converting to a final library size of 2e9. The converted libraries were also sequenced by NGS to confirm that they had a distribution of >1e8 heavy chains and >1e5 light chains. The heavy and light chains were cloned separately, but the library size was such that any starting clone with an occurrence frequency higher than 5e-4 would have a native pair present in the resulting library. The library was confirmed to contain all human native V genes. 2 × 10⁻⁶ 9 The transform was obtained during the library transformation.

[0291] A set of 25 homologs was selected for toxin-Fc-AviTag and toxin-10 His-AviTag fusions. Fc tags and 10 His tags (His-His-His-His-His-His; SEQ ID NO: 326) were provided to assist in toxin purification, and AviTag was provided to enable site-directed biotinylation to magnetic beads for autosomal phase panning. When expression was induced in HEK293 cells, 21 of the 25 toxins produced measurable titers. All recombinant toxins were then exposed to serum from highly immunized donors, and 15 of the 21 toxins, including several species that the donors had never been directly immunized before, showed strong reactivity to the serum. These results demonstrate that (1) highly immunized subjects are receptive to these specific toxins, and (2) toxins are produced in a form recognizable by donor antibodies.

[0292] Isolation of a broadly neutralizing anti-alpha-neurotoxin antibody. The inventors demonstrated that their approach technique can successfully extract the crucial proof of principle for a fully human, broadly neutralizing antibody (Centi-D09) against alpha-neurotoxin, one of the "Big Four" toxins in toxins. The molecular basis of the broadly neutralizing antitoxin antibody was established through in vivo defense models in multiple snake species, crystallography of bnAb in complex with alpha-neurotoxin from multiple snake species, and dynamic and biophysical characterization of bnAb.

[0293] In certain embodiments, the novelty of the embodiments described herein lies in three features of the next-generation antitoxin: 1) the inventors' antibody supply source; 2) technological development; and 3) storage of the final product.

[0294] The technology described herein utilizes a novel, diverse immunotherapy library collected from middle-aged men who have undergone dose-escalating autoimmunization (over 700 boosts and 200 raw bites) over the past 19 years from 17 diverse snake species, including the Western diamond, Mojave, coral snake, black mamba, Western green mamba, Eastern green mamba, Jameson's mamba, Thai cobra, Egyptian cobra, forest cobra, water cobra, Cape cobra, Indian krait, round-tailed krait, coastal taipan, PNG taipan, tiger snake, and Eastern brown snake. These antibodies are safe and effective in the inventors' middle-aged men and offer advantages compared to humanizing antibodies from animal sources or naive human libraries. Furthermore, their single-domain nature may result in a shortened half-life compared to complete human IgG. Repeated injections are justified due to their small size, which allows for renal excretion if not adequately neutralized with a single dose.

[0295] Robotic Interspecies Phage Panning. The technique described herein utilizes a unique next-generation sequencing (NGS) to guide the amplification and tracking of an antibody variable domain repertoire from blood samples taken before and 28 days after toxicology immunization in male subjects, followed by deep sequencing and antibody phage display to enable tracing downstream toxin-binding factors back to the source blood samples, thereby enabling the generation of an immunotherapy library. This antibody library contains a high concentration of high-affinity mature antitoxin antibodies selected for a broad range of snakes and demonstrating the ability to protect against high doses of lethal snake venom in vivo. In addition, our robotic homolog cross-panning protocol enables the rapid isolation of broad-spectrum antitoxin neutralizing antibodies. We have already demonstrated the value of this combination of library and robotic cross-panning in the recovery of the in vivo protective long-chain neurotoxin bnAb Centi-D9.

[0296] Heat-stabilized lyophilized antibodies. Standard antitoxins require continuous refrigeration at 2–8°C to maintain stability. This poses a significant challenge to arid villages, often inhabited by locally endemic toxic species, which may lack the infrastructure to properly store antitoxins. In certain cases, the antibodies described herein are evaluated in relation to the room-temperature stability of lyophilized or high-concentration formulation antitoxins. This application describes heat-stabilized antibodies with less stringent storage requirements and longer shelf lives, which would greatly expand the types of medical centers where they can be stored and made available, enabling on-site deployment of life-saving drugs at poisoning sites.

[0297] The experiments described herein involved (1) identifying and characterizing a pool of cross-reactive, high-affinity antitoxin antibody candidates; (2) exploring mechanisms of action through epitope characterization from the entire toxin pool; and (3) determining specific defense patterns through in vivo challenge in mice.

[0298] The inventors have demonstrated the existence of broad-spectrum neutralizing antibodies (bnAbs) against snake venom. Centi-D9, a broad-spectrum, ultra-high-affinity antibody against long-chain neurotoxins, was able to neutralize long-chain neurotoxins from cobras, taipans, and mambas, and provide in vivo protection against them.

[0299] The inventors also demonstrated that the biophysical properties of the most promising clone, Centi-D9, can be optimized by mutations in Centi-D9 residues into germline residues at specific locations; and by the removal of biochemical modification sites such as the N-linked glycosylation motif NS. The thermal stability and aggregation of these variants were tested for the modified clones based on the parent Centi-D9 shown in Table 29. Table 21B demonstrates that these modified clones retain affinity for several long-chain neurotoxins derived from the venom of snakes of the family Elapidae. [Table 29]

[0300] Identification of antibody candidates against three toxins: The inventors identified 193, 97, and 190 antibody clones against neurotoxin, phospholipase A2 (PLA2), and E-dendrotoxin, respectively, by ELISA screening (Figure 12). Long-chain neurotoxins were screened by ELISA. Thirteen clones were selected by ranked offrate of periplasmic extracts (PPE) using Octet HTX, then reformatted and expressed IgG. K values ​​ranging from 2.75e-09 to 2.63e-10 and 3.3e08 to 1.75e09 were found for various neurotoxins. D High-resolution kinetic data were generated for two candidates, Centi-D09 and Centi-B11, for three of four toxins (long-chain neurotoxins from taipan, cobra, mamba, and krait), each possessing a specific characteristic. Evaluated at both 25°C and 37°C using Octet HTX and BiaCORE 8K, Centi-D9 is a very high-affinity (picomolecular) bnAb for the three snake species (taipan, cobra, and mamba) (490 pM, 37 pM, and 74 pM, respectively). For dendrotoxins, 10 of the first 190 epsilon(E) dendrotoxin-positive clones were also ELISA-positive for A and D-dendrotoxins. Six of these 10, which had the broadest spectrum against the three different dendrotoxins, were reformatted to IgG. The binding kinetics of reformatted dendrotoxin antibody candidates were characterized, and one clone (Centi-DTX-B03) showed strong binding activity to dendrotoxins from four different species, demonstrating the neutralizing width of its antibody. Crystallographic evidence of the complex was observed for one clone. Anti-PLA2 and anti-Kunitz clones were identified and characterized for their width by ELISA.

[0301] The following were achieved: 1) generation of kinetic data for each antibody tested against each toxin; 2) reformatting of antibody candidates to whole human IgG; and 3) characterization of the binding affinity of the reformatted IgG antibodies. Since the PLA2 enzyme was found to be toxic to mammalian cells expressing recombinant proteins, the inventors adapted their protocol to the use of biotinylated PLA2 isolated from whole snake venom, confirmed to be active by phospholipase enzyme assay, and used for panning. As a result, this product was successfully used for panning and screening.

[0302] The dynamics and binding mechanism of the antibody Centi-D9 (SNEURO_P01_D09). Crystal structures were generated to define its binding orientation and parameters (Figure 13). Four crystals mapped Centi-D09 to complexes with long-chain neurotoxins from the krait (Bungarus caeruleus) (called alpha-bungarotoxin in this species), the black mamba (Dendroaspis polylepsis), the coastal taipan (Oxyuranus scutatellus), and the Cape cobra (Naja nivea). The crystal structures demonstrated that Centi-D9 binds to the long-chain neurotoxins from all four species in a similar manner, thereby preventing the binding of long-chain neurotoxins to nicotinic acetylcholine receptors (nAChRs). This is demonstrated by the binding footprints of Centi-D9 and nAChR, which almost completely overlap on the surface of long-chain neurotoxins (Figure 13F). Both Centi-D9 and Centi-B11 (SNEURO_P01_B11) bind to toxins from these four elapidated species, and Centi-D9 exhibits favorable kinetics compared to Centi-B11 in whole Biacore800 and Octet HTX kinetic assays. The crystal structure of Centi-D9 in complex with these long-chain neurotoxins clearly demonstrates a conserved binding mode / motif across these diverse toxins (there are no pairs among them with more than 65% sequence identity): the VH CDR3 of Centi-D9 contacts at least one residue in “finger” loop I and at least one residue in “finger” loop II.

[0303] Centi-D9 in in vivo challenge studies using mouse models. Using a Centi-D9 mouse model for in vivo challenge with toxin injection, the lethal dose ranges for four elapidated species (Table 30) were established, and the functional activity and broad-spectrum efficacy of Centi-D9 against recombinant toxins and whole snake venom in vivo were determined. [Table 30]

[0304] 1 LD50 Derived from experiments on four total snake venoms tested during dose-range studies to establish the LD100. LD100 .

[0305] Mice exposed to lethal doses of Thai cobra, black mamba, and Cape cobra venom were completely protected from death by broad-spectrum neutralizing activity after pretreatment with Centi-D9, which significantly exceeded expectations in terms of protection against the total toxin challenge as a single antibody (as opposed to exposure to the cocktail) (Figure 15). Based on abundant evidence indicating that long-chain neurotoxins are the primary drivers of lethality in most Elapidae snakes, and therefore the crucial basis for a universal antitoxin cocktail in Elapidae snakes, we chose to focus on Elapidae. Centi-D09 was demonstrated to be a broad-spectrum neutralizing antibody that provides in vivo protection against the total toxins of six Elapidae species (Figure 16).

[0306] The presence of Centi-D09 provided evidence that broad-spectrum neutralizing antibodies against homologous snake toxins exist and can be isolated as the basis for universal antitoxins. Furthermore, the identification of Centi-D09 demonstrates that broad-spectrum neutralizing antibodies against snake venom toxins from various species have been successfully isolated from highly immune subjects with a history of repeated onset of humoral immune responses to various such toxins.

[0307] Broad-spectrum neutralizing antitoxins may require combinations of agents that neutralize separate components found in two major classes of venomous snakes: Elapidae and Viperidae (Table 31). For neurotoxic Elapidae snakes, venom virulence is driven by long-chain neurotoxins (3FT) and PLA2. Anti-PLA2 bnAbs from a panel of candidate binding factors can be combined with the anti-long-chain neurotoxin bnAb Centi-D9 to create a two-component cocktail that provides protection against Elapidae snakes. [Table 31]

[0308] 1 The toxins listed are the primary toxins of the result, and their relative abundances are indicated by the number of checkmarks, although their abundances can vary considerably among the specific toxins of individual snake species from both families.

[0309] The single 3FT bnAb, Centi-D09, was found to act against the black mamba (Dendroaspis polylepsis), Cape cobra (Naja nivea), and Thai cobra (Naja kaouthia). Further investigation revealed that Centi-D09 also acts against other members of the Elapidae family: the Egyptian cobra (Naja haje), Indian cobra (Naja naja), and king cobra (Ophiphagus hannah). Centi-D09 was further shown to bind to the 3FTX long-chain neurotoxin in toxins obtained from the tiger snake (notechis scutatus scutatus), the spotted sea snake (laticauda colubrina), the western green mamba (dendoapsis viridis), the Indonesian poison cobra (naja sputatrix), the inland taipan (oxyuranus microlepidotus), and the eastern brown snake (pseudonaja textilis) by Gator BLI assay (Figure 16).

[0310] In some cases, Centi-D09 can be combined with an anti-PLA2 inhibitor. For example, Centi-D09 can be combined with valesprazib, a small molecule PLA2 inhibitor for coastal taipan. Alternatively, the small molecule inhibitor can be replaced with a PLA2 bnAb. In other cases, Centi-D09 can be combined with an antibody against a dendrotoxin (kunitz-like peptide) specific to the genus dendroaspis.

[0311] Production of recombinant toxins. The EXPi-293 mammalian expression system is used for the expression of toxin antigens. Toxin proteins are purified using Protein G DYNABEADS™, biotinylated, and quality control is performed for biotinylation and binding to control antibodies. Some toxins are difficult to produce using this method because the toxin is lethal to protein-producing cells. In these cases (e.g., PLA2), the whole snake venom is obtained and fractionated using HPLC to obtain purified toxin for biotinylation.

[0312] Panning of phage libraries against toxin antigens. Immunological library (2 × 10⁻⁶) 9 The mixture is heated at 72°C for 10 minutes to deselect for Protein G DYNABEADS (INVITROGEN®), M-280 streptavidin DYNABEADS (INVITROGEN®), histone (Sigma) from calf thymus, human IgG (Sigma), ssDNA-biotin NNK (Integrated DNA Technologies), and DNA-biotin NNK (Integrated DNA Technologies). The library is then sequentially panned against the toxic antigens captured by M-280 streptavidin DYNABEADS (INVITROGEN®) using an automated protocol with Kingfisher FLEX (THERMO FISHER SCIENTIFIC®). Selected phages are eluted from the beads with acid and neutralized using Tris-HCl pH 7.9 (TEKNOVA®). ER2738 cells were infected with a neutralized phage pool at a ratio of 1:10 with OD600=0.5. After incubation at 37°C and 100 rpm for 40 minutes, the phage pool was centrifuged and incubated overnight on agar at 30°C with antibiotic selection. Rescued phages were precipitated with PEG and subjected to three additional rounds of automated soluble phase panning. The resulting antibody clones were screened by ELISA for binding activity to recombinant proteins or whole snake venom.

[0313] Preliminary ELISA-positive kinetic characterization by SPR. The kinetic characteristics of a pool of ELISA-positive antibodies against all snake venoms and fractionated toxins are characterized using the CARTERRA® LSA (CSLA), a high-throughput SPR platform. Each antibody is tested against all toxins or fractionated toxins for kinetic characterization and cross-reactivity binding. Antibodies are further characterized for thermal stability and aggregation using fluorescence and light scattering techniques with Unchained Labs (UNcle). Selection of clones with the fastest on-rate (ka) and slowest off-rate (kd) minimizes trauma to victims, as the toxin should be rapidly neutralized and not released.

[0314] The binding kinetics of our ELISA-positive antibody candidates are tested using CLSA. The data suggest that these candidates bind to the snake venom they pan. Their binding affinity is characterized to determine their potential functionality in neutralizing target toxins. Since cross-reactive molecules are used to achieve broad-spectrum antitoxin treatment, it is determined whether any of our candidates can bind to ≥2 toxin species or toxins. SPR technology enables phage display, and kinetic data (ka [association rate], kd [dissociation rate], K) are obtained from bacterial supernatant expressing our antibody candidates without purification. Currently expressed using a marked scFv phagemide, phage display is possible. D It becomes possible to accurately determine the [equilibrium dissociation constant]). Using CARTERRA® software, which uses the maximum resonance ([A]Rmax) (KD=[A]Rmax / Req) of the antigen at a specific concentration bound to scFv on the chip, divided by resonance equilibrium, K DThe CLSA platform advantageously tests ≥4 specific antigens on 384 antibody candidates in ≤18 hours (h). This high-throughput technology determines the relative affinity of all candidates to the panned original toxin and ≥20 different whole toxins / fractionated peptide toxins in <7 days. The candidate with the highest affinity (<10 nM) to the toxin is reformatted as IgG in our mammalian expression system. The IgG is purified and tested for thermal stability and aggregation tendency. scFv expressed by the bacterial cell line ER2738 is captured using anti-V5 antibody coupled to the CLSA chip. The Centivax phagemide vector has a C-terminal V5 tag inserted, thereby enabling downstream scFv capture. Whole snake venom and toxins are flowed onto any captured clone using an 8-point dilution scheme at a maximum concentration of 3 μM and a minimum concentration of 450 pM to determine binding kinetics and the presence of cross-reactivity. Anti-Her2 scFv is captured as a negative control for all toxins.

[0315] The CLSA platform is a versatile, high-throughput monoclonal antibody characterization method that combines patented continuous-flow microfluidics with array SPR detection, providing high-volume data generation in reduced time and consumable costs. We are the first to use this technique to determine antibody binding or cross-reactivity to snake species, as most snake bite laboratories lack access to such SPR equipment.

[0316] For low-affinity binding factors, millions of variants of these clones can be generated by CDRH3 mutagenesis and light chain shuffling using marked-affinity matured POLISH technology. These sets of clones are screened for higher affinity (<10 nM) using CLSA. If poor cross-reactivity occurs, NGS data from the panning round can be used to express specific clones enriched through panning but not present during the initial ELISA screening, and these can be tested with ELISA and CARTERRA® in addition to the initially positive ones.

[0317] Generate kinetic data for each antibody tested against each toxin. <10nM K D The acceptable affinity is determined by the on-rate [ka] of 10⁴ ¹ / Ms to 10⁵ ¹ / Ms and the off-rate [kd] of 10⁻⁴ ¹ / s to 10⁻⁶ ¹ / s. The clone with the highest affinity (<10 nM) for all toxins or toxins ≥1, and / or the cross-reactive antibody (indicated by binding to species ≥1), is selected for IgG reformatting.

[0318] Reformatting of antibody candidates to human IgG. Identified antibody candidates are reformatted from scFv to full human IgG by PCR. The goal is to bind to and neutralize toxins, rather than to induce a downstream inflammatory immune response, so IgG backlines with minimal effector function (e.g., LALA mutations) are used. Designed primers are fitted with suitable ends for cloning to the mammalian expression vector pTT5. Restriction sites suitable for the pTT5 vector are added by PCR to each scFv candidate for the heavy-chain V gene and light-chain V gene (previously characterized by SS). The resulting products are digested and cloned into DH5-alpha bacterial cells. Proper cloning is confirmed by Sanger sequencing (Eton Bioscience). Cloned DNA is midiprepped using a commercially available kit (ZYMO RESEARCH®) and transiently transfected into HEK293 cells for mammalian expression. The supernatant from each cell is collected 5 days after transfection. IgG is purified and quantified using Octet QK with Protein A at a minimum expression level of 50 μg / mL. Expression purity is analyzed by SDS-PAGE. The pTT5 vector is used for reformatting and expression of antibody candidates.

[0319] Stability and kinetic characterization of IgG antibody candidates. The binding affinity of reformatted antibody candidates in IgG format at CLSA is determined against previously tested toxins / peptides. While scFv contains the antibody domain that determines antigen binding, affinity may change when complexed with the constant region of complete IgG. Complete IgG contains two binding sites compared to one on scFv or Fab, so binding efficiency may be increased due to avidity. The stability of IgG candidates is characterized using UNchained Labs (UNcle). UNcle uses dynamic light scattering, static light scattering, and fluorescence to determine the protein's melting temperature, aggregation tendency, and isothermal stability, which are critical to determining the overall stability of the IgG candidate and the quality of downstream antitoxins.

[0320] IgG is thermally unstable and prone to aggregation. To correct this, the IgG complementarity-determining region (CDR) can be mutated and grafted onto a more thermally stable human germline framework. By altering the scaffold to which the CDR is loaded, the CDR can be maintained, creating a more thermally stable molecule. This can be achieved by using a known thermally stable human germline scaffold containing the CDR of a selected antibody candidate, synthesized by IDT or Twist and cloned into the inventors' pTT5 vector. The binding dynamics are retested to determine the retention of binding.

[0321] IgG can be bound with the same or higher affinity as when bound in scFv format (<10 nM), which has improved thermal stability and aggregation tendency due to UNcle. Only IgG that is a 95% monomer with a melting temperature (Tm) of 68°C or higher and a KD of <10 nM will be used for subsequent purposes.

[0322] The antibody / antibody cocktail of the present invention can be lyophilized. The formulation for lyophilization may contain histidine, methionine, and polysorbate 80, for example, 50 mM histidine, 20 mM methionine, 0.05% polysorbate 80 (w / v), pH 6.8. The lyophilized formulation may further contain a combination of bulking agents / lyophilization stabilizers (including, but not limited to, sorbitol, mannitol, sucrose, dextrose, and glycine). The lyophilized dosage formulation is stored at 50°C and analyzed at regular time points over 28 days by the following methods: appearance, SEC-HPLC, optical density, pH, dynamic light scattering, T melting, T aggregation, particulate matter, water content, or any combination thereof.

[0323] In certain cases, if a common set of lyophilization parameters allows all antibodies to be processed stably together, multiple antibodies can be lyophilized in a single cocktail. Alternatively, the antibodies can be lyophilized separately, the freeze-dried powders mixed by weight, and the combined powder reconstituted for downstream analysis.

[0324] In one embodiment, a cocktail of monoclonal antibodies against lethal toxins in snake venom, i.e., antibodies providing broad protection against vipers and cobras, is described. Each of these antibodies is individually optimized to broadly neutralize the entire family of polymorphic toxins (e.g., a single antibody capable of neutralizing dendrotoxins from cobras worldwide). By combining several broad-spectrum anti-toxin antibodies, snake bite lethality can be eliminated by neutralizing the most potent toxins present in the venom of the world's most dangerous snakes. This unique combination of high-potency and broad-spectrum antibodies and cocktail strategy makes it possible to create snake bite drugs that enable significantly faster treatment without requiring identification of the venom-injecting species. Furthermore, lyophilization of the antibody cocktail creates a drug that is ultrastable and can be stored at ambient temperature for extended periods. Two-chamber auto-injectors for use by untrained individuals or in field rescue settings, which can be easily carried and used by travelers, medical personnel, or military personnel in areas where deadly snake bites are a serious concern, are also described.

[0325] Preferred embodiments of the present invention have been shown and described herein, but it will be apparent to those skilled in the art that such embodiments are provided merely as examples. A great many variations, alterations, and substitutions will now come to mind to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein can be used when carrying out the invention. The scope of the present invention is defined by the following claims, and methods and structures within the scope of these claims, as well as their equivalents, are intended to be covered thereby. In certain embodiments, for example, the following are provided: (Item 1) A method for treating a subject suffering from poison injection, comprising the step of administering to the subject a composition comprising an effective amount of a universal antitoxin comprising one or more antibodies or one or more antigen-binding fragments described herein. (Item 2) The method according to item 1, wherein the injection of venom is carried out by one or more species of snakes. (Item 3) The aforementioned poison injection affected Boiga irregularis (southern giant snake), Boiga cyanea (green giant snake), Boiga dendrophila (mangrove snake), Dispholidus typus (boomslang), Salvadora grahamiae (mountain patchnose snake), Spalerosophis diadema (crowned snake), Tantilla nigriceps (plains blackhead snake), Thelotornis capensis (southern twig snake), Thelotornis kirtlandii (northern twig snake), Toxicodryas blandingii (branding tree snake), Trimorphodon lambda (Sonoran lyre snake), Amphiesma stolatum (buff kaleback), Natrix tessellate (dice snake), and Rhabdophis subminiatus (Red-necked Japanese keelback), Rhabdophis tigrinus (Japanese keelback), Thamnophis elegans (Western terrestrial garter snake), Thamnophis sirtalis (Common garter snake), Ahaetulla nasuta (Long-nosed whip snake), Atractaspis bibronii (Bibrons' burrowing asp), Atractaspis dahomeyensis (Dahomey's burrowing asp), Atractaspis engaddensis (Palestinian mole snake), Atractaspis microlepidota (Small-scaled burrowing asp), Malpolon monspessulanus (Montpellier snake), Acanthophis antarcticus (Common death adder), Aipysurus laevis (Olive brown sea snake), Aipysurus duboisii (Dubois's sea snake), Austrelaps superbus (lowland pit viper), Cryptophis nigrescens (small ice snake), Demansia olivacea (olive whip snake), Emydocephalus annulatus (spotted sea snake), Furinatristis (Stevens' striped snake), Hydrophis melanocephalus (black-headed sea snake), Hydrophis curtus (spiny sea snake), Hydrophis gracilis (slender sea snake), Hydrophis elegans (elegant sea snake), Hydrophis jerdonii (corn-nosed sea snake), Hydrophis klossi (slangor sea snake), Hydrophis peronii (horned sea snake), Hydrophis belcheri (Belcher's sea snake), Hydrophis stricticollis (Bengal sea snake), Hydrophis major (olive-headed sea snake), Hydrophis stokesii (large-headed sea snake), Hydrophis melanosoma (black-banded robust sea snake), Hydrophis hardwickii (dorsal-ventral sea snake), Hydrophis cyanocinctus (spotted sea snake), Hydrophis spiralis (narrow-banded sea snake), Hydrophis nigrocinctus (black-banded sea snake), Hydrophis platurus (black-backed sea snake), Hydrophis ornatus (black-spotted sea snake), Hydrophis viperinus (venomous sea snake), Hydrophis schistosus (wart-tailed sea snake), Notechis scutatus (mainland tiger snake), Oxyuranus scutellatus (coastal taipan), Oxyuranus temporalis (central range taipan), Pseudechis australis (margath snake), Pseudechis butleri (Butler's black snake), Pseudechis Colletti (Colette's black snake), Pseudechis guttatus (blue-bellied black snake), Pseudechis papuanus (Papua black snake), Pseudechis porphyriacus (red-bellied black snake), Pseudonaja affinis (Dugait's black snake), Pseudonajaguttata (Speckled Brown Snake), Pseudonaja inframacula (Peninsula Brown Snake), Pseudonaja nuchalis (Western Brown Snake), Pseudonaja textilis (Eastern Brown Snake), Tropidechis carinatus (Clarence River Snake), Aspidelaps lubricus (Cape Coral Snake), Aspidelaps scutatus (Shield-nose Snake), Bungarus fasciatus (Round-tailed Krait), Bungarus caeruleus (Indoor Krait), Bungarus candidus (Blue Krait), Bungarus flaviceps (Redhead Crate), Bungarus multicinctus (Krait), Dendroaspis viridis (Western Green Mamba), Dendroaspis angusticeps (Eastern Green Mamba), Dendroaspis jamesoni (Jameson's Mamba), Dendroaspis polylepis (Black Mamba), Elapsoidea sundevallii (Sandebaru African Garter Snake), Hemachatus haemachatus (Lincalus), Laticauda colubrina (Blue-spotted Sea Snake), Laticauda laticaudata (Broad-sea Sea Snake), Laticauda semifasciata (Erabu Sea Snake), Micrurus obscurus (Bolivian Coral Snake), Micrurus frontalis (Southern Coral Snake), Micrurus alleni (Allen's Coral Snake), Micrurus altirostris (Uruguayan Coral Snake), Micrurus clarki (Clark's Coral Snake), Micrurus corallinus (Painted Coral Snake), Micrurus Micrurus distans (Western Mexican coral snake), Micrurus dumerilii (Dumeril's coral snake), Micrurus fulvius (Eastern coral snake), Micrurus hemprichii (Hemprich's coral snake), Micrurusibiboboca (Kaachinga coral snake), Micrurus lemniscatus (South American coral snake), Micrurus mipartitus (Red-tailed coral snake), Micrurus mosquitensis (Costa Rican coral snake), Micrurus multifasciatus (Striped coral snake), Micrurus nigrocinctus (Central American coral snake), Micrurus pyrrhocryptus (Argentine coral snake), Micrurus spixii (Amazonian coral snake), Micrurus surinamensis (Aquatic coral snake), Micrurus tener (Texas coral snake), Micrurus tschudii (Desert coral snake), Naja siamensis (Indochinese spitting cobra), Naja annulata (Ringed water cobra), Naja annulifera (Long-nosed cobra), Naja ashei (Giant spitting cobra), Naja atra (Taiwanese cobra), Naja christyi (Congos cobra), Naja haje (Egyptian cobra), Naja kaouthia (Thai cobra), Naja katiensis (Western African spitting cobra), Naja melanoleuca (Forest cobra), Naja mossambica (Mozambican spitting cobra), Naja naja (Indian cobra), Naja nigricollis (Black-necked spitting cobra), Naja nivea (Cape cobra), Naja nubiae (Nubian spitting cobra), Naja oxiana (Caspian cobra), Naja pallida (Red spitting cobra), Naja philippinensis (Northern Philippine cobra), Naja samarensis (Samanthus cobra), Naja Sputatrix (Indonesian spitting cobra), Naja sumatrana (Sumatran spitting cobra), Ophiophagus hannah (King cobra), Walterinnesia aegyptia (Western desert black cobra), Homalopsis buccata (Linnaeus's water snake), Myrrophis chinensis (Chinese mud snake), SubsessorBocourti (Bocourt's water snake), Azemiops feae (cobra viper), Agkistrodon bilineatus (patterned pit viper), Agkistrodon contortrix (American pit viper), Agkistrodon piscivorus (swamp pit viper), Agkistrodon taylori (castellana), Agkistrodon laticinctus (broadband copperhead), Atropoides picadoi (Picado's jumping pit viper), Bothriechis lateralis (sidestriped palm pit viper), Bothriechis nigroviridis (black-spotted palm pit viper), Bothriechis schlegelii (eyelash palm pit viper), Bothrops diporus (chacolance head), Bothrops erythromelas (curting lance head), Bothrops Bothrops insularis (Golden Lancehead Viper), Bothrops jararaca (Jararaka), Bothrops neuwiedi (Neuwiedi Lancehead), Bothrops pauloensis (Blackfaced Lancehead), Bothrops asper (Terciopero), Bothrops atrox (Caicasa), Bothrops ayerbei (Ayerbei Lancehead), Bothrops caribbaeus (Saint Lucia Lancehead), Bothrops jararacussu (Jararacus), Bothrops lanceolatus (Martiny Lancehead), Bothrops leucurus (Whitetail Lancehead), Bothrops moojeni (Brazil Lancehead), Bothrops alternatus (Urutu), Bothrops cotiara (Cotiara), Bothrops fonsecai (Fonseca Lancehead), Bothrops itapetiningae (São Paulo Lance Head), Bothrops taeniatus (Speckled Forest Pit Viper), Bothropsmattogrossensis (Matogrosso otter), Calloselasma rhodostoma (Malayan pit viper), Cerrophidion godmani (Godman Mountains viper), Cerrophidion sasai (Costa Rican mountain viper), Crotalus viridis (prairie rattlesnake), Crotalus atrox (Western diamondback rattlesnake), Crotalus a damanteus (Eastern diamondback rattlesnake), Crotalus basiliscus (Mexican west coast rattlesnake), Crotalus catalinensis (Santa Catalina Island rattlesnake), Crotalus cerastes (sideborne rattlesnake), Crotalus Cerberus (Arizona black rattlesnake), Crotalus durissus (South American rattlesnake), Crotalus enyo (Baja California rattlesnake), Crotalus horridus (wood rattlesnake), Crotalus lepidus (Mottled rock rattlesnake), Crotalus mitchellii (San Lucan spotted rattlesnake), Crotalus molossus (Northern black-tailed rattlesnake), Crotalus oreganus (North Pacific rattlesnake), Crotalus pricei (Western twin-spotted rattlesnake), Crotalus pusillus (Tacita Rattlesnake), Crotalus ravus (Mexican Pygmy Rattlesnake), Crotalus ruber (Red Diamond Rattlesnake), Crotalus scutulatus (Mojave Rattlesnake), Crotalus simus (Central American Rattlesnake), Crotalus tigris (Tiger Rattlesnake), Crotalus totonacus (Totonacan Rattlesnake), Crotalus tzabcan (Yucatan Neotropical Rattlesnake), Crotalus willardi (Arizona Ridgenose Rattlesnake), Crotalus Pyrrhus (Southwestern Speckled Rattlesnake), Crotalus vegrandis (Uracoan Rattlesnake), Deinagkistrodon acutus (Hyappoda), Gloydius intermedius (Intermediate Viper), Gloydius blomhoffii (pit viper), Gloydius brevicaudus (snake pit viper), Gloydius halys (Siberian pit viper), Gloydius shedaoensis (snake island pit viper), Gloydius ussuriensis (ussuri pit viper), Hypnale hypnale (knobby pit viper), Lachesismelanocephala (blackhead bushmaster), Lachesis muta (Atlantic bushmaster), Ovophis okinavensis (Okinawan pit viper), Porthidium nasutum (tropical rainforest pit viper), Porthidium ophryomegas (slender pit viper), Protobothrops elegans (Sakishima pit viper), Protobothrops flavoviridis (Okinawa pit viper), Protobothrops mangshanensis (Manshan pit viper), Protobothrops mucrosquamatus (Taiwanese pit viper), Protobothrops tokarensis (Tokara pit viper), Sistrurus catenatus (Masasogai), Sistrurus miliarius (pygmy rattlesnake), Trimeresurus stejnegeri (Taiwanese green pit viper), Trimeresurus albolabris (white-lipped pit viper), Trimeresurus erythrurus (Burmese pit viper), Trimeresurus gramineus (Japanese pit viper), Trimeresurus labialis (Nicobar bamboo pit viper), Trimeresurus macrops (large-eyed pit viper), Trimeresurus malabaricus (Malabar pit viper), Trimeresurus popeiorum (Pope pit viper), Trimeresurus purpureomaculatus (mangrove pit viper), Trimeresurus sumatranus (Sumatran pit viper), Tropidolaemus wagleri (Wagler's pit viper), Metlapilcoatlus mexicanus (Mexican jumping pit viper), Metlapilcoatlus nummifer (Central American jumping pit viper), Atheris squamigera (lizard bush viper), Bitis arietans (puff adder), Bitis Atropos (Cape Mountain adder), Bitis caudalis (horned puff adder), Bitis cornuta (Western branched horn adder), BitisGabonica (Central African Gabonese viper), Bitis nasicornis (rhinoceros adder), Bitis parviocula (Ethiopian adder), Bitis rhinoceros (West African Gabonese viper), Causus rhombeatus (common night adder), Cerastes cerastes (horned viper), Cerastes gasperettii (Arabian horned viper), Cerastes vipera (Saharan sand viper), Daboia palaestinae (Palestinian viper), Daboia russelii (Russell's viper), Daboia siamensis (Eastern Russell's viper), Daboia mauritanica (Moorish viper), Echis ocellatus (West African carpet viper), Echis carinatus (sawtooth viper), Echis coloratus (painted saw-toothed snake), Echis pyramidum (Egyptian saw-toothed snake), Macrovipera schweizeri (Milo's viper), Macrovipera lebetinus (Levant viper), Montivipera bornmuelleri (Lebanese mountain viper), Montivipera latifii (Latific viper), Montivipera raddei (Armenian mountain viper), Montivipera xanthine (Ottoman viper), Pseudocerastes fieldi (Field's horned viper), Pseudocerastes persicus (Persian horned viper), Vipera ammodytes (European nose-nosed viper), Vipera aspis (Aspian viper), Vipera berus (European viper), Vipera latastei (Latast's viper), Vipera lotievi (Caucasian wild viper), Vipera seoanei (Basque viper), Vipera ursinii (Wild viper), Diadophis punctatus (Ring-necked viper), Hydrodynastes gigas (Water cobra), HypsiglenaThe method described in item 1 or 2, which is one or more snakes selected from the group consisting of torquata (night snake), Hypsiglena jani (San Luis Stoposi night snake), Leptodeira ashmeadii, Philodryas olfersii (Liechtenstein green racer), and combinations thereof. (Item 4) The method described in item 3, wherein the venom injection is from (a) a sawtooth snake, water moccasin, lancehead, rattlesnake, Russell's viper, puff adder, or a combination thereof, or (b) a krait (Bungarus caeruleus), black mamba (Dendroaspis polylepsis), coastal taipan (Oxyuranus scutatellus), Cape cobra (Naja nivea), or a combination thereof. (Item 5) The method according to item 4, wherein the subject is administered one antibody, two antibodies or antigen-binding fragments, three antibodies or antigen-binding fragments, four antibodies or antigen-binding fragments, five antibodies or antigen-binding fragments, six antibodies or antigen-binding fragments, seven antibodies or antigen-binding fragments, eight antibodies or antigen-binding fragments, nine antibodies or antigen-binding fragments, ten antibodies or antigen-binding fragments, or more antibodies or antigen-binding fragments. (Item 6) The method according to any one of items 1 to 5, wherein the one or more antibodies or antigen-binding fragments include, are derived from, or are a combination thereof, IgG, IgM, IgE, IgA, or IgD. (Item 7) The method according to any one of items 4 to 6, wherein the one or more antibodies or antigen-binding fragments include a monoclonal antibody, a graft antibody, a chimeric antibody, a human antibody, a humanized antibody, or a combination thereof. (Item 8) The antigen-binding fragment is Fab, Fab', F(ab') 2 Variable fragment (Fv), triabody, tetrabody, minibody, binocular F(ab') 2 , triple specificity F(ab') 2 The method described in any one of items 1 to 7, including diabodies, bispecific diabodies, single-chain variable fragments (scFv), scFv-Fc, Fab-Fc, VHH, bispecific scFv, or combinations thereof. (Item 9) The method according to any one of items 1 to 8, further comprising the step of administering an additional therapy or drug to the subject. (Item 10) The method according to item 9, wherein the additional therapy or drug comprises a PLA2 inhibitor. (Item 11) The method according to item 10, wherein the PLA2 inhibitor includes valesprazib, methylvalesprazib, or a combination thereof. (Item 12) The method according to any one of items 1 to 10, wherein the administration comprises one or more injections. (Item 13) The method according to any one of items 1 to 12, wherein the one or more antibodies or antigen-binding fragments comprises a VH CDR1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 2. (Item 14) The method according to any one of items 1 to 13, wherein the one or more antibodies or antigen-binding fragments comprises a VH CDR2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 3. (Item 15) The method according to any one of items 1 to 14, wherein the one or more antibodies or antigen-binding fragments include a variable heavy chain (VH) complementarity-determining region 3 (CDR3) having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 1. (Item 16) The method according to any one of items 1 to 15, wherein the one or more antibodies or antigen-binding fragments comprises a variable light chain (VL) CDR1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 4. (Item 17) The method according to any one of items 1 to 16, wherein the one or more antibodies or antigen-binding fragments comprises a VL CDR2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 5. (Item 18) The method according to any one of items 1 to 17, wherein the one or more antibodies or antigen-binding fragments comprises a VL CDR3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 6. (Item 19) The method according to any one of items 1 to 18, wherein the one or more antibodies or antigen-binding fragments comprises FW-H1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 8. (Item 20) The method according to any one of items 1 to 19, wherein the one or more antibodies or antigen-binding fragments comprises FW-H2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 9. (Item 21) The method according to any one of items 1 to 20, wherein the one or more antibodies or antigen-binding fragments comprises FW-H3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 10. (Item 22) The method according to any one of items 1 to 21, wherein the one or more antibodies or antigen-binding fragments comprises FW-H4 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 11. (Item 23) The method according to any one of items 1 to 22, wherein the one or more antibodies or antigen-binding fragments comprises FW-L1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 12. (Item 24) The method according to any one of items 1 to 23, wherein the one or more antibodies or antigen-binding fragments comprises FW-L2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 13. (Item 25) The method according to any one of items 1 to 24, wherein the one or more antibodies or antigen-binding fragments comprises FW-L3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 14. (Item 26) The method according to any one of items 1 to 25, wherein the one or more antibodies or antigen-binding fragments comprises FW-L4 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 15. (Item 27) The method according to any one of items 1 to 26, wherein the one or more antibodies or antigen-binding fragments comprises a VH having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 16. (Item 28) The method according to any one of items 1 to 27, wherein the one or more antibodies or antigen-binding fragments comprises a VL having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 17. (Item 29) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 1, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 68, and VL CDR3 of SEQ ID NO: 84. (Item 30) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 224 and VL of SEQ ID NO: 249. (Item 31) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 18, VH CDR2 of SEQ ID NO: 34, VH CDR3 of SEQ ID NO: 2, VL CDR1 of SEQ ID NO: 47, VL CDR2 of SEQ ID NO: 69, and VL CDR3 of SEQ ID NO: 85. (Item 32) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 225 and VL of SEQ ID NO: 250. (Item 33) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 3, VL CDR1 of SEQ ID NO: 48, VL CDR2 of SEQ ID NO: 70, and VL CDR3 of SEQ ID NO: 86. (Item 34) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 228 and VL of SEQ ID NO: 251. (Item 35) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 18, VH CDR2 of SEQ ID NO: 34, VH CDR3 of SEQ ID NO: 2, VL CDR1 of SEQ ID NO: 49, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 87. (Item 36) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 227 and VL of SEQ ID NO: 252. (Item 37) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 18, VH CDR2 of SEQ ID NO: 34, VH CDR3 of SEQ ID NO: 2, VL CDR1 of SEQ ID NO: 50, VL CDR2 of SEQ ID NO: 70, and VL CDR3 of SEQ ID NO: 88. (Item 38) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 228 and VL of SEQ ID NO: 253. (Item 39) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 19, VH CDR2 of SEQ ID NO: 35, VH CDR3 of SEQ ID NO: 4, VL CDR1 of SEQ ID NO: 51, VL CDR2 of SEQ ID NO: 72, and VL CDR3 of SEQ ID NO: 89. (Item 40) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 229 and VL of SEQ ID NO: 254. (Item 41) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 5, VL CDR1 of SEQ ID NO: 52, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 90. (Item 42) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 230 and VL of SEQ ID NO: 255. (Item 43) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 20, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 6, VL CDR1 of SEQ ID NO: 53, VL CDR2 of SEQ ID NO: 73, and VL CDR3 of SEQ ID NO: 91. (Item 44) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 231 and VL of SEQ ID NO: 256. (Item 45) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 231 and VL of SEQ ID NO: 257. (Item 46) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 21, VH CDR2 of SEQ ID NO: 36, VH CDR3 of SEQ ID NO: 7, VL CDR1 of SEQ ID NO: 54, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 92. (Item 47) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 232 and VL of SEQ ID NO: 258. (Item 48) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 22, VH CDR2 of SEQ ID NO: 37, VH CDR3 of SEQ ID NO: 8, VL CDR1 of SEQ ID NO: 55, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 84. (Item 49) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 233 and VL of SEQ ID NO: 259. (Item 50) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 1, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 68, and VL CDR3 of SEQ ID NO: 84. (Item 51) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 224 and VL of SEQ ID NO: 249. (Item 52) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 23, VH CDR2 of SEQ ID NO: 38, VH CDR3 of SEQ ID NO: 9, VL CDR1 of SEQ ID NO: 56, VL CDR2 of SEQ ID NO: 75, and VL CDR3 of SEQ ID NO: 93. (Item 53) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 234 and VL of SEQ ID NO: 280. (Item 54) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 234 and VL of SEQ ID NO: 281. (Item 55) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 24, VH CDR2 of SEQ ID NO: 39, VH CDR3 of SEQ ID NO: 10, VL CDR1 of SEQ ID NO: 57, VL CDR2 of SEQ ID NO: 76, and VL CDR3 of SEQ ID NO: 94. (Item 56) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 235 and VL of SEQ ID NO: 282. (Item 57) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 25, VH CDR2 of SEQ ID NO: 40, VH CDR3 of SEQ ID NO: 11, VL CDR1 of SEQ ID NO: 58, VL CDR2 of SEQ ID NO: 77, and VL CDR3 of SEQ ID NO: 95. (Item 58) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 236 and VL of SEQ ID NO: 283. (Item 59) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 40, VH CDR3 of SEQ ID NO: 11, VL CDR1 of SEQ ID NO: 59, VL CDR2 of SEQ ID NO: 78, and VL CDR3 of SEQ ID NO: 96. (Item 60) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 237 and VL of SEQ ID NO: 284. (Item 61) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 237 and VL of SEQ ID NO: 285. (Item 62) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 27, VH CDR2 of SEQ ID NO: 40, VH CDR3 of SEQ ID NO: 11, VL CDR1 of SEQ ID NO: 60, VL CDR2 of SEQ ID NO: 79, and VL CDR3 of SEQ ID NO: 97. (Item 63) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 238 and VL of SEQ ID NO: 286. (Item 64) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 238 and VL of SEQ ID NO: 287. (Item 65) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 55, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 84. (Item 66) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 239 and VL of SEQ ID NO: 288. (Item 67) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 55, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 98. (Item 68) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 240 and VL of SEQ ID NO: 289. (Item 69) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 3, VL CDR1 of SEQ ID NO: 61, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 99. (Item 70) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 241 and VL of SEQ ID NO: 270. (Item 71) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 62, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 100. (Item 72) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 239 and VL of SEQ ID NO: 271. (Item 73) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 63, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 92. (Item 74) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 239 and VL of SEQ ID NO: 272. (Item 75) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 80, and VL CDR3 of SEQ ID NO: 101. (Item 76) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 242 and VL of SEQ ID NO: 273. (Item 77) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 64, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 102. (Item 78) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 243 and VL of SEQ ID NO: 274. (Item 79) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 65, VL CDR2 of SEQ ID NO: 81, and VL CDR3 of SEQ ID NO: 103. (Item 80) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 243 and VL of SEQ ID NO: 275. (Item 81) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 54, VL CDR2 of SEQ ID NO: 82, and VL CDR3 of SEQ ID NO: 104. (Item 82) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 244 and VL of SEQ ID NO: 276. (Item 83) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 51, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 105. (Item 84) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 239 and VL of SEQ ID NO: 277. (Item 85) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 29, VH CDR2 of SEQ ID NO: 42, VH CDR3 of SEQ ID NO: 13, VL CDR1 of SEQ ID NO: 65, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 92. (Item 86) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 245 and VL of SEQ ID NO: 278. (Item 87) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 30, VH CDR2 of SEQ ID NO: 43, VH CDR3 of SEQ ID NO: 14, VL CDR1 of SEQ ID NO: 51, VL CDR2 of SEQ ID NO: 72, and VL CDR3 of SEQ ID NO: 106. (Item 88) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 246 and VL of SEQ ID NO: 279. (Item 89) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 31, VH CDR2 of SEQ ID NO: 44, VH CDR3 of SEQ ID NO: 15, VL CDR1 of SEQ ID NO: 66, VL CDR2 of SEQ ID NO: 69, and VL CDR3 of SEQ ID NO: 107. (Item 90) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 247 and VL of SEQ ID NO: 280. (Item 91) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 32, VH CDR2 of SEQ ID NO: 45, VH CDR3 of SEQ ID NO: 16, VL CDR1 of SEQ ID NO: 67, VL CDR2 of SEQ ID NO: 83, and VL CDR3 of SEQ ID NO: 108. (Item 92) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 248 and VL of SEQ ID NO: 281. (Item 93) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 24, VH CDR2 of SEQ ID NO: 39, VH CDR3 of SEQ ID NO: 10, VL CDR1 of SEQ ID NO: 57, VL CDR2 of SEQ ID NO: 76, and VL CDR3 of SEQ ID NO: 94. (Item 94) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 235 and VL of SEQ ID NO: 282. (Item 95) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 349, VH CDR2 of SEQ ID NO: 371, and VH CDR3 of SEQ ID NO: 327. (Item 96) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 350, VH CDR2 of SEQ ID NO: 372, and VH CDR3 of SEQ ID NO: 328. (Item 97) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 351, VH CDR2 of SEQ ID NO: 373, and VH CDR3 of SEQ ID NO: 329. (Item 98) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 352, VH CDR2 of SEQ ID NO: 374, and VH CDR3 of SEQ ID NO: 330. (Item 99) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 353, VH CDR2 of SEQ ID NO: 375, and VH CDR3 of SEQ ID NO: 331. (Item 100) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 354, VH CDR2 of SEQ ID NO: 376, and VH CDR3 of SEQ ID NO: 332. (Item 101) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 355, VH CDR2 of SEQ ID NO: 377, and VH CDR3 of SEQ ID NO: 333. (Item 102) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 356, VH CDR2 of SEQ ID NO: 378, and VH CDR3 of SEQ ID NO: 334. (Item 103) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 357, VH CDR2 of SEQ ID NO: 379, and VH CDR3 of SEQ ID NO: 335. (Item 104) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 358, VH CDR2 of SEQ ID NO: 380, and VH CDR3 of SEQ ID NO: 336. (Item 105) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 359, VH CDR2 of SEQ ID NO: 381, and VH CDR3 of SEQ ID NO: 337. (Item 106) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 360, VH CDR2 of SEQ ID NO: 382, ​​and VH CDR3 of SEQ ID NO: 338. (Item 107) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 361, VH CDR2 of SEQ ID NO: 383, and VH CDR3 of SEQ ID NO: 339. (Item 108) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 362, VH CDR2 of SEQ ID NO: 384, and VH CDR3 of SEQ ID NO: 340. (Item 109) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 363, VH CDR2 of SEQ ID NO: 385, and VH CDR3 of SEQ ID NO: 341. (Item 110) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 364, VH CDR2 of SEQ ID NO: 386, and VH CDR3 of SEQ ID NO: 342. (Item 111) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 365, VH CDR2 of SEQ ID NO: 387, and VH CDR3 of SEQ ID NO: 343. (Item 112) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 366, VH CDR2 of SEQ ID NO: 388, and VH CDR3 of SEQ ID NO: 344. (Item 113) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 367, VH CDR2 of SEQ ID NO: 389, and VH CDR3 of SEQ ID NO: 345. (Item 114) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 368, VH CDR2 of SEQ ID NO: 390, and VH CDR3 of SEQ ID NO: 346. (Item 115) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 369, VH CDR2 of SEQ ID NO: 391, and VH CDR3 of SEQ ID NO: 347. (Item 116) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 370, VH CDR2 of SEQ ID NO: 392, and VH CDR3 of SEQ ID NO: 348. (Item 117) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 393, VL CDR2 of SEQ ID NO: 405, and VL CDR3 of SEQ ID NO: 417. (Item 118) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 394, VL CDR2 of SEQ ID NO: 406, and VL CDR3 of SEQ ID NO: 418. (Item 119) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 395, VL CDR2 of SEQ ID NO: 407, and VL CDR3 of SEQ ID NO: 419. (Item 120) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 396, VL CDR2 of SEQ ID NO: 408, and VL CDR3 of SEQ ID NO: 420. (Item 121) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 397, VL CDR2 of SEQ ID NO: 409, and VL CDR3 of SEQ ID NO: 421. (Item 122) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 398, VL CDR2 of SEQ ID NO: 410, and VL CDR3 of SEQ ID NO: 422. (Item 123) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 399, VL CDR2 of SEQ ID NO: 411, and VL CDR3 of SEQ ID NO: 423. (Item 124) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 400, VL CDR2 of SEQ ID NO: 412, and VL CDR3 of SEQ ID NO: 424. (Item 125) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 401, VL CDR2 of SEQ ID NO: 413, and VL CDR3 of SEQ ID NO: 425. (Item 126) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 402, VL CDR2 of SEQ ID NO: 414, and VL CDR3 of SEQ ID NO: 426. (Item 127) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 403, VL CDR2 of SEQ ID NO: 415, and VL CDR3 of SEQ ID NO: 427. (Item 128) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VL CDR1 of SEQ ID NO: 404, VL CDR2 of SEQ ID NO: 416, and VL CDR3 of SEQ ID NO: 428. (Item 129) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 429 and one VL of any of SEQ ID NOs: 501 to 512. (Item 130) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 430 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 131) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 431 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 132) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 432 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 133) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 433 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 134) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 434 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 135) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 435 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 136) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 436 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 137) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 437 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 138) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 438 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 139) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 439 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 140) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 440 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 141) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 441 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 142) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 442 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 143) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 443 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 144) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 444 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 145) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 445 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 146) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 446 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 147) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 447 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 148) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 448 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 149) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 449 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 150) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments comprise the VH of SEQ ID NO: 450 and the VL of any one of SEQ ID NOs: 501 to 512. (Item 151) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 513 and VL of SEQ ID NO: 518. (Item 152) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 514 and VL of SEQ ID NO: 519. (Item 153) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 515 and VL of SEQ ID NO: 520. (Item 154) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 516 and VL of SEQ ID NO: 521. (Item 155) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 517 and VL of SEQ ID NO: 522. (Item 156) The method according to any one of items 1 to 28, wherein the one or more antibodies or antigen-binding fragments include VH of SEQ ID NO: 523 and VL of SEQ ID NO: 524. (Item 157) A universal antitoxin composition comprising one or more antitoxin antibodies or antigen-binding fragments that selectively bind to one or more toxins from one or more species of snakes. (Item 158) A universal antitoxin composition as described in item 157, comprising a population of antitoxin antibodies or antigen-binding fragments, each comprising one or more antibodies or antigen-binding fragments. (Item 159) The universal antitoxin composition described in item 158, wherein the population of antitoxin antibodies or antigen-binding fragments comprises two antibodies or antigen-binding fragments, three antibodies or antigen-binding fragments, four antibodies or antigen-binding fragments, five antibodies or antigen-binding fragments, six antibodies or antigen-binding fragments, seven antibodies or antigen-binding fragments, eight antibodies or antigen-binding fragments, nine antibodies or antigen-binding fragments, ten antibodies or antigen-binding fragments, or more antibodies or antigen-binding fragments. (Item 160) A universal antitoxin composition according to any one of items 157 to 159, wherein the one or more antibodies or antigen-binding fragments comprises, derived from, or a combination thereof, IgG, IgM, IgE, IgA, or IgD. (Item 161) A universal antitoxin composition according to any one of items 157 to 160, wherein the one or more antibodies or antigen-binding fragments comprises a monoclonal antibody, a graft antibody, a chimeric antibody, a human antibody, a humanized antibody, or a combination thereof. (Item 162) The antigen-binding fragment is Fab, Fab', F(ab') 2 Variable fragment (Fv), triabody, tetrabody, minibody, binocular F(ab') 2 , triple specificity F(ab') 2 A universal antitoxin composition according to any one of items 157 to 161, comprising a diabody, a bispecific diabody, a single-chain variable fragment (scFv), scFv-Fc, Fab-Fc, VHH, bispecific scFv, or a combination thereof. (Item 163) A universal antitoxin composition according to any one of items 157 to 162, wherein the one or more antibodies or antigen-binding fragments comprises a VH CDR3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 1. (Item 164) A universal antitoxin composition according to any one of items 157 to 163, wherein the one or more antibodies or antigen-binding fragments comprises a VH CDR1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 2. (Item 165) A universal antitoxin composition according to any one of items 157 to 164, wherein the one or more antibodies or antigen-binding fragments comprises a VH CDR2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 3. (Item 166) A universal antitoxin composition according to any one of items 157 to 165, wherein the one or more antibodies or antigen-binding fragments comprises a VL CDR1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 4. (Item 167) A universal antitoxin composition according to any one of items 157 to 166, wherein the one or more antibodies or antigen-binding fragments comprises a VL CDR2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 5. (Item 168) A universal antitoxin composition according to any one of items 157 to 167, wherein the one or more antibodies or antigen-binding fragments comprises a VL CDR3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 6. (Item 169) A universal antitoxin composition according to any one of items 157 to 168, comprising FW-H1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 8. (Item 170) A universal antitoxin composition according to any one of items 157 to 169, wherein the one or more antibodies or antigen-binding fragments comprises FW-H2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 9. (Item 171) A universal antitoxin composition according to any one of items 157 to 170, wherein the one or more antibodies or antigen-binding fragments comprises FW-H3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 10. (Item 172) A universal antitoxin composition according to any one of items 157 to 171, wherein the one or more antibodies or antigen-binding fragments comprises FW-H4 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 11. (Item 173) A universal antitoxin composition according to any one of items 157 to 172, wherein the one or more antibodies or antigen-binding fragments comprises FW-L1 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 12. (Item 174) A universal antitoxin composition according to any one of items 157 to 173, wherein the one or more antibodies or antigen-binding fragments comprises FW-L2 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any one of the sequences in Table 13. (Item 175) A universal antitoxin composition according to any one of items 157 to 174, wherein the one or more antibodies or antigen-binding fragments comprises FW-L3 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 14. (Item 176) A universal antitoxin composition according to any one of items 157 to 175, wherein the one or more antibodies or antigen-binding fragments comprises FW-L4 having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 15. (Item 177) A universal antitoxin composition according to any one of items 157 to 176, wherein the one or more antibodies or antigen-binding fragments comprises a VH having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 16. (Item 178) A universal antitoxin composition according to any one of items 157 to 177, wherein the one or more antibodies or antigen-binding fragments comprises a VL having an amino acid sequence that is at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of the sequences in Table 17. (Item 179) The one or more antibodies or antigen-binding fragments mentioned above are found in Boiga irregularis (southern giant snake), Boiga cyanea (green giant snake), Boiga dendrophila (mangrove snake), Dispholidus typus (boomslang), Salvadora grahamiae (mountain patchnose snake), Spalerosophis diadema (crowned snake), Tantilla nigriceps (plains blackhead snake), Thelotornis capensis (southern twig snake), Thelotornis kirtlandii (northern twig snake), Toxicodryas blandingii (branding tree snake), Trimorphodon lambda (Sonoran ryar snake), Amphiesma stolatum (buff kaleback), Natrix tessellate (dice snake), and Rhabdophis subminiatus (Red-necked Japanese keelback), Rhabdophis tigrinus (Japanese keelback), Thamnophis elegans (Western terrestrial garter snake), Thamnophis sirtalis (Common garter snake), Ahaetulla nasuta (Long-nosed whip snake), Atractaspis bibronii (Bibrons' burrowing asp), Atractaspis dahomeyensis (Dahomey's burrowing asp), Atractaspis engaddensis (Palestinian mole snake), Atractaspis microlepidota (Small-scaled burrowing asp), Malpolon monspessulanus (Montpellier snake), Acanthophis antarcticus (Common death adder), Aipysurus laevis (Olive brown sea snake), Aipysurus duboisii (Dubois's sea snake), Austrelaps superbus (lowland pit viper), Cryptophis nigrescens (small ice snake), Demansia olivacea (olive whip snake), EmydocephalusHydrophis annulatus (spotted sea snake), Furina tristis (Stevens' striped snake), Hydrophis melanocephalus (black-headed sea snake), Hydrophis curtus (spiny sea snake), Hydrophis gracilis (slender sea snake), Hydrophis elegans (elegant sea snake), Hydrophis jerdonii (corn-nosed sea snake), Hydrophis klossi (slangor sea snake), Hydrophis peronii (horned sea snake), Hydrophis belcheri (Belcher's sea snake), Hydrophis stricticollis (Bengal sea snake), Hydrophis major (olive-headed sea snake), Hydrophis stokesii (large-headed sea snake), Hydrophis melanosoma (black-banded robust sea snake), Hydrophis hardwickii (dorsal-ventral sea snake), Hydrophis cyanocinctus (spotted sea snake), Hydrophis spiralis (narrow-banded sea snake), Hydrophis nigrocinctus (black-banded sea snake), Hydrophis platurus (black-backed sea snake), Hydrophis ornatus (black-spotted sea snake), Hydrophis viperinus (venomous sea snake), Hydrophis schistosus (wart sea snake), Notechis scutatus (mainland tiger snake), Oxyuranus scutellatus (coastal taipan), Oxyuranus temporalis (central range taipan), Pseudechis australis (margath snake), Pseudechis Butler's black snake, Pseudechis colletti, Pseudechis guttatus, Pseudechis papuanus, Pseudechis porphyriacus, Pseudechis porphyriacusaffinis (Dugait), Pseudonaja guttata (Speckled Brown Snake), Pseudonaja inframacula (Peninsula Brown Snake), Pseudonaja nuchalis (Western Brown Snake), Pseudonaja textilis (Eastern Brown Snake), Tropidechis carinatus (Clarence River Snake), Aspidelaps lubricus (Cape Coral Snake), Aspidelaps scutatus (Shield-nose Snake), Bungarus fasciatus (Round-tailed Krait), Bungarus caeruleus (Indoor Krait), Bungarus candidus (Blue Krait), Bungarus flaviceps (Redhead Crate), Bungarus multicinctus (Krait), Dendroaspis viridis (Western Green Mamba), Dendroaspis angusticeps (Eastern Green Mamba), Dendroaspis jamesoni (Jameson's Mamba), Dendroaspis polylepis (Black Mamba), Elapsoidea sundevallii (Sandebaru African Garter Snake), Hemachatus haemachatus (Lincalus), Laticauda colubrina (Blue-spotted Sea Snake), Laticauda laticaudata (Broad-sea Sea Snake), Laticauda semifasciata (Erabu Sea Snake), Micrurus obscurus (Bolivian Coral Snake), Micrurus frontalis (Southern Coral Snake), Micrurus alleni (Allen's Coral Snake), Micrurus altirostris (Uruguayan Coral Snake), Micrurus clarki (Clark's Coral Snake), Micrurus corallinus (Painted Coral Snake), Micrurus distans (Western Mexican coral snake), Micrurus dumerilii (Dumeril's coral snake), Micrurus Micrurus fulvius (Eastern coral snake), Micrurus hemprichii (Hemprich's coral snake), Micrurus ibiboboca (Kaachinga coral snake), Micrurus lemniscatus (South American coral snake), Micrurus mipartitus (Red-tailed coral snake), Micrurus mosquitensis (Costa Rican coral snake), Micrurus multifasciatus (Striped coral snake), Micrurus nigrocinctus (Central American coral snake), Micrurus pyrrhocryptus (Argentine coral snake), Micrurus spixii (Amazonian coral snake), Micrurus surinamensis (Aquatic coral snake), Micrurus tener (Texas coral snake), Micrurus tschudii (Desert coral snake), Naja siamensis (Indochinese spitting cobra), Naja Naja annulata (Ringed Water Cobra), Naja annulifera (Long-nosed Cobra), Naja ashei (Giant Poisonous Cobra), Naja atra (Taiwanese Cobra), Naja christyi (Conglomerate Water Cobra), Naja haje (Egyptian Cobra), Naja kaouthia (Thai Cobra), Naja katiensis (Western African Poisonous Cobra), Naja melanoleuca (Forest Cobra), Naja mossambica (Mozambican Poisonous Cobra), Naja naja (Indian Cobra), Naja nigricollis (Black-necked Poisonous Cobra), Naja nivea (Cape Cobra), Naja nubiae (Nubian Poisonous Cobra), Naja oxiana (Caspian Cobra), Naja pallida (Red Poisonous Cobra), Naja philippinensis (Northern Philippine Cobra), Naja Samarensis (Samara cobra), Naja sputatrix (Indonesian spitting cobra), Naja sumatrana (Sumatran spitting cobra), Ophiophagus hannah (King cobra), Walterinnesia aegyptia (Western desert cobra), Homalopsisbuccata (Linnaeus's water snake), Myrrophis chinensis (Chinese mud snake), Subsessor bocourti (Bocourt's water snake), Azemiops feae (cobra viper), Agkistrodon bilineatus (patterned pit viper), Agkistrodon contortrix (American pit viper), Agkistrodon piscivorus (swamp pit viper), Agkistrodon taylori (castellana), Agkistrodon laticinctus (broadband copperhead), Atropoides picadoi (Picado's jumping pit viper), Bothriechis lateralis (sidestriped palm pit viper), Bothriechis nigroviridis (black spotted palm pit viper), Bothriechis schlegelii (eyelash palm pit viper), Bothrops diporus (chacolance head), Bothrops erythromelas (Curtin Lancehead), Bothrops insularis (Golden Lancehead Viper), Bothrops jararaca (Jararaka), Bothrops neuwiedi (Neuwiedi Lancehead), Bothrops pauloensis (Blackfaced Lancehead), Bothrops asper (Terciopero), Bothrops atrox (Kaikasa), Bothrops ayerbei (Ayerbei Lancehead), Bothrops caribbaeus (Saint Lucia Lancehead), Bothrops jararacussu (Jararacus), Bothrops lanceolatus (Martiny Lancehead), Bothrops leucurus (Whitetail Lancehead), Bothrops moojeni (Brazil Lancehead), Bothrops alternatus (Urutu), Bothrops cotiara (Cotiara), Bothrops fonsecai (Lance Head of Fonseca), Bothrops itapetiningae (Lance Head of São Paulo), Bothropstaeniatus (Speckled Forest Pit Viper), Bothrops mattogrossensis (Matogrosso Lanzen Otter), Calloselasma rhodostoma (Murray Pit Viper), Cerrophidion godmani (Godman Mountains Pit Viper), Cerrophidion sasai (Costa Rican Mountains Pit Viper), Crotalus viridis (Prairie Rattlesnake), Crotalus atrox (Western Diamondback Rattlesnake), Crotalus adamanteus (Eastern Diamondback Rattlesnake), Crotalus basiliscus (Mexico West Coast Rattlesnake), Crotalus catalinensis (Santa Catalina Island Rattlesnake), Crotalus cerastes (Leafhopper Rattlesnake), Crotalus Cerberus (Arizona Black Rattlesnake), Crotalus durissus (South American Rattlesnake), Crotalus enyo (Baja California rattlesnake), Crotalus horridus (wood rattlesnake), Crotalus lepidus (Mottled rock rattlesnake), Crotalus mitchellii (San Lucan spotted rattlesnake), Crotalus molossus (Northern black-tailed rattlesnake), Crotalus oreganus (North Pacific rattlesnake), Crotalus pricei (Western twin-spotted rattlesnake), Crotalus pusillus (Tacita Landa sky rattlesnake), Crotalus ravus (Mexican pygmy rattlesnake), Crotalus ruber (Red diamond rattlesnake), Crotalus scutulatus (Mojave rattlesnake), Crotalus simus (Central American rattlesnake), Crotalus tigris (Tiger rattlesnake), Crotalus totonacus (Totonakan rattlesnake), Crotalus tzabcan (Yucatan Neotropical rattlesnake), Crotalus willardi (Arizona ridgenose rattlesnake), Crotalus Pyrrhus (Southwestern speckled rattlesnake), CrotalusVegrandis (Uracoan rattlesnake), Deinagkistrodon acutus (Hyappoda), Gloydius intermedius (Intermediate pit viper), Gloydius blomhoffii (Pit viper), Gloydius brevicaudus (Long-nosed pit viper), Gloydius halys (Siberian pit viper), Gloydius shedaoensis (Snake Island pit viper), Gloydius ussuriensis (Ussuri pit viper), Hypnale hypnale (Bumpy-nosed pit viper), Lachesis melanocephala (Blackhead bushmaster), Lachesis muta (Atlantic bushmaster), Ovophis okinavensis (Rubber pit viper), Porthidium nasutum (Tropical rainforest hognose pit viper), Porthidium ophryomegas (Slender hognose pit viper), Protobothrops elegans (Sakishima pit viper), Protobothrops flavoviridis (Okinawa pit viper), Protobothrops mangshanensis (Manshan pit viper), Protobothrops mucrosquamatus (Taiwan pit viper), Protobothrops tokarensis (Tokara pit viper), Sistrurus catenatus (Masasogai pit viper), Sistrurus miliarius (Pygmy rattlesnake), Trimeresurus stejnegeri (Taiwan green pit viper), Trimeresurus albolabris (White-lipped green pit viper), Trimeresurus erythrurus (Burmese green pit viper), Trimeresurus gramineus (Green pit viper), Trimeresurus labialis (Nicobar bamboo pit viper), Trimeresurus macrops (Large-eyed pit viper), Trimeresurus malabaricus (Malabar pit viper), Trimeresurus Popeiorum (Pop's pit viper), Trimeresurus purpureomaculatus (mangrove pit viper), Trimeresurus sumatranus (Sumatran pit viper), TropidolaemusWagleri (Wagler's pit viper), Metlapilcoatlus mexicanus (Mexican jumping pit viper), Metlapilcoatlus nummifer (Central American jumping pit viper), Atheris squamigera (lizard bush viper), Bitis arietans (puff adder), Bitis Atropos (Cape mountain adder), Bitis caudalis (horned puff adder), Bitis cornuta (Western branched horned adder), Bitis gabonica (Central African Gabon viper), Bitis nasicornis (rhinoceros adder), Bitis parviocula (Ethiopian adder), Bitis rhinoceros (West African Gabon viper), Causus rhombeatus (common night adder), Cerastes cerastes (horned viper), Cerastes gasperettii (Arabian horned viper), Cerastes vipera (Saharan sand pit viper), Daboia palaestinae (Palestinian viper), Daboia russelii (Russell's viper), Daboia siamensis (Eastern Russell's viper), Daboia mauritanica (Moorish viper), Echis ocellatus (West African carpet viper), Echis carinatus (sawtooth viper), Echis coloratus (painted sawtooth viper), Echis pyramidum (Egyptian sawtooth viper), Macrovipera schweizeri (Milos viper), Macrovipera lebetinus (Levant viper), Montivipera bornmuelleri (Lebanese mountain viper), Montivipera latifii (latific viper), Montivipera raddei (Armenian mountain viper), Montivipera xanthine (Ottoman viper), Pseudocerastes fieldi (Field horned viper), Pseudocerastes persicus (Persian horned viper), Vipera ammodytes (European nose-nosed viper), Vipera aspis (Aspian viper), Vipera berus (European viper), Vipera latastei (Latast's viper), Vipera lotievi (Caucasian field viper), Vipera seoanei (Basque viper), Vipera ursinii (Field viper), Diadophis punctatus (Ring-necked viper), Hydrodynastes gigas (Water cobra), Hypsiglena A universal antitoxin composition according to any one of items 157 to 178, which binds to one or more epitopes on toxins from one or more snakes selected from the group consisting of torquata (night snake), Hypsiglena jani (San Luis toposi night snake), Leptodeira ashmeadii, Philodryas olfersii (Liechtenstein green racer), and combinations thereof. (Item 180) A universal antitoxin composition according to any one of items 157 to 179, wherein the one or more antibodies or antigen-binding fragments bind to one or more epitopes on a toxin from one or more snakes selected from the group consisting of sawtooth snakes, water mocassin snakes, lancehead snakes, rattlesnakes, Russell's vipers, puff adders, and combinations thereof. (Item 181) A universal antitoxin composition according to any one of items 157 to 179, wherein one or more antibodies or antigen-binding fragments bind to toxins from sawtooth snakes, water mocassin, lancehead snakes, rattlesnakes, Russell's vipers, and puff adders. (Item 182) A universal antitoxin composition according to any one of items 157 to 179, wherein the one or more antibodies or antigen-binding fragments bind to one or more epitopes on a toxin from one or more snakes selected from the group consisting of the krait (Bungarus caeruleus), the black mamba (Dendroaspis polylepsis), the coastal taipan (Oxyuranus scutatellus), the Cape cobra (Naja nivea), and combinations thereof. (Item 183) A universal antitoxin composition according to any one of items 157 to 179, wherein one or more antibodies or antigen-binding fragments bind to toxins from the krait (Bungarus caeruleus), black mamba (Dendroaspis polylepsis), coastal taipan (Oxyuranus scutatellus), and Cape cobra (Naja nivea). (Item 184) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 1, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 68, and VL CDR3 of SEQ ID NO: 84. (Item 185) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 224 and VL of SEQ ID NO: 249. (Item 186) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 18, VH CDR2 of SEQ ID NO: 34, VH CDR3 of SEQ ID NO: 2, VL CDR1 of SEQ ID NO: 47, VL CDR2 of SEQ ID NO: 69, and VL CDR3 of SEQ ID NO: 85. (Item 187) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 225 and VL of SEQ ID NO: 250. (Item 188) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 3, VL CDR1 of SEQ ID NO: 48, VL CDR2 of SEQ ID NO: 70, and VL CDR3 of SEQ ID NO: 86. (Item 189) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 228 and VL of SEQ ID NO: 251. (Item 190) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 18, VH CDR2 of SEQ ID NO: 34, VH CDR3 of SEQ ID NO: 2, VL CDR1 of SEQ ID NO: 49, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 87. (Item 191) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 227 and VL of SEQ ID NO: 252. (Item 192) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 18, VH CDR2 of SEQ ID NO: 34, VH CDR3 of SEQ ID NO: 2, VL CDR1 of SEQ ID NO: 50, VL CDR2 of SEQ ID NO: 70, and VL CDR3 of SEQ ID NO: 88. (Item 193) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 228 and VL of SEQ ID NO: 253. (Item 194) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 19, VH CDR2 of SEQ ID NO: 35, VH CDR3 of SEQ ID NO: 4, VL CDR1 of SEQ ID NO: 51, VL CDR2 of SEQ ID NO: 72, and VL CDR3 of SEQ ID NO: 89. (Item 195) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 229 and VL of SEQ ID NO: 254. (Item 196) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 5, VL CDR1 of SEQ ID NO: 52, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 90. (Item 197) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 230 and VL of SEQ ID NO: 255. (Item 198) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 20, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 6, VL CDR1 of SEQ ID NO: 53, VL CDR2 of SEQ ID NO: 73, and VL CDR3 of SEQ ID NO: 91. (Item 199) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 231 and VL of SEQ ID NO: 256. (Item 200) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 231 and VL of SEQ ID NO: 257. (Item 201) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 21, VH CDR2 of SEQ ID NO: 36, VH CDR3 of SEQ ID NO: 7, VL CDR1 of SEQ ID NO: 54, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 92. (Item 202) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 232 and VL of SEQ ID NO: 258. (Item 203) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 22, VH CDR2 of SEQ ID NO: 37, VH CDR3 of SEQ ID NO: 8, VL CDR1 of SEQ ID NO: 55, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 84. (Item 204) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 233 and VL of SEQ ID NO: 259. (Item 205) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 1, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 68, and VL CDR3 of SEQ ID NO: 84. (Item 206) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 224 and VL of SEQ ID NO: 249. (Item 207) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 23, VH CDR2 of SEQ ID NO: 38, VH CDR3 of SEQ ID NO: 9, VL CDR1 of SEQ ID NO: 56, VL CDR2 of SEQ ID NO: 75, and VL CDR3 of SEQ ID NO: 93. (Item 208) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 234 and VL of SEQ ID NO: 280. (Item 209) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 234 and VL of SEQ ID NO: 281. (Item 210) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 24, VH CDR2 of SEQ ID NO: 39, VH CDR3 of SEQ ID NO: 10, VL CDR1 of SEQ ID NO: 57, VL CDR2 of SEQ ID NO: 76, and VL CDR3 of SEQ ID NO: 94. (Item 211) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 235 and VL of SEQ ID NO: 282. (Item 212) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 25, VH CDR2 of SEQ ID NO: 40, VH CDR3 of SEQ ID NO: 11, VL CDR1 of SEQ ID NO: 58, VL CDR2 of SEQ ID NO: 77, and VL CDR3 of SEQ ID NO: 95. (Item 213) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 236 and VL of SEQ ID NO: 283. (Item 214) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 40, VH CDR3 of SEQ ID NO: 11, VL CDR1 of SEQ ID NO: 59, VL CDR2 of SEQ ID NO: 78, and VL CDR3 of SEQ ID NO: 96. (Item 215) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 237 and VL of SEQ ID NO: 284. (Item 216) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 237 and VL of SEQ ID NO: 285. (Item 217) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 27, VH CDR2 of SEQ ID NO: 40, VH CDR3 of SEQ ID NO: 11, VL CDR1 of SEQ ID NO: 60, VL CDR2 of SEQ ID NO: 79, and VL CDR3 of SEQ ID NO: 97. (Item 218) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 238 and VL of SEQ ID NO: 286. (Item 219) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 238 and VL of SEQ ID NO: 287. (Item 220) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 55, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 84. (Item 221) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 239 and VL of SEQ ID NO: 288. (Item 222) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 55, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 98. (Item 223) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 240 and VL of SEQ ID NO: 289. (Item 224) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 17, VH CDR2 of SEQ ID NO: 33, VH CDR3 of SEQ ID NO: 3, VL CDR1 of SEQ ID NO: 61, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 99. (Item 225) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 241 and VL of SEQ ID NO: 270. (Item 226) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 62, VL CDR2 of SEQ ID NO: 74, and VL CDR3 of SEQ ID NO: 100. (Item 227) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 239 and VL of SEQ ID NO: 271. (Item 228) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 63, VL CDR2 of SEQ ID NO: 71, and VL CDR3 of SEQ ID NO: 92. (Item 229) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ ID NO: 239 and VL of SEQ ID NO: 272. (Item 230) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments include VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 80, and VL CDR3 of SEQ ID NO: 101. (Item 231) A universal antitoxin composition according to any one of items 157 to 183, wherein the one or more antibodies or antigen-binding fragments comprise VH of SEQ...

Claims

1. An antibody or antigen-binding fragment that is a broad-spectrum neutralizing antibody against two or more members of the family of three-finger toxins (3FTx), comprising VH CDR1 of SEQ ID NO: 28, VH CDR2 of SEQ ID NO: 41, VH CDR3 of SEQ ID NO: 12, VL CDR1 of SEQ ID NO: 46, VL CDR2 of SEQ ID NO: 80, and VL CDR3 of SEQ ID NO:

101.

2. The broad-spectrum neutralizing antibody or antigen-binding fragment according to claim 1, wherein the 3FTx is a long-chain neurotoxin.

3. A universal antitoxin composition comprising the broad-spectrum neutralizing antibody or antigen-binding fragment described in Claim 1.

4. A universal antitoxin composition for use in a method of treating a subject suffering from poison injection, the method comprising the step of administering the universal antitoxin composition to the subject.

5. The universal antitoxin composition for use according to claim 4, wherein the venom injection is carried out by one or more species of snakes.

6. The broad-spectrum neutralizing antibody or antigen-binding fragment according to claim 1, wherein the antibody neutralizes alpha-bungarotoxin (krait), alpha-elapitoxin (mamba), pseudonajatoxin (brown snake), alpha-cobratoxin (cobra), toxin B (king cobra), or a combination thereof.

7. The broad-spectrum neutralizing antibody or antigen-binding fragment according to claim 2, wherein the antibody or antigen-binding fragment broadly neutralizes toxins from three or more long-chain neurotoxins.

8. The broad-spectrum neutralizing antibody or antigen-binding fragment according to claim 1, wherein the antibody or antigen-binding fragment comprises a VH having the amino acid sequence of SEQ ID NO:

242.

9. The broad-spectrum neutralizing antibody or antigen-binding fragment according to claim 1, wherein the antibody or antigen-binding fragment comprises a VL having the amino acid sequence of SEQ ID NO:

273.

10. Use of the universal antitoxin composition according to claim 3 in the manufacture of a pharmaceutical for treating a subject suffering from poison injection.

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