Human immunodeficiency virus neutralizing antibody formulations and uses thereof

Abstract

The present application provides a human immunodeficiency virus neutralizing antibody preparation and its use, specifically, the preparation comprises (a) a therapeutically effective amount of HIV neutralizing antibody; (b) a buffer with a concentration of 10-30 mmol / L; (c) a stabilizer; and (d) a surfactant, wherein the pH of the liquid preparation is 5.5-6.5. The preparation of the present application can not only effectively reduce the chemical degradation reaction rate of human immunodeficiency virus neutralizing antibody, improve the physical and chemical stability of the antibody, prolong the shelf life of the product, and reduce the number of drug bottles, but also can eliminate or reduce the side reactions of the injection site of the patient, and improve the drug comfort of the patient.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and more specifically to a human immunodeficiency virus neutralizing antibody preparation and its uses. Background Technology

[0002] AIDS, or Acquired Immunodeficiency Syndrome, is a highly fatal infectious disease caused by the human immunodeficiency virus (HIV). HIV invades the human body, destroying the immune system and gradually eroding the body's resistance to various diseases, ultimately leading to death. Currently, there is no vaccine to prevent it, nor are there any effective drugs or treatments to cure the disease.

[0003] The HIV-neutralizing antibody of this invention was developed by the team of Academician Nussenzweig, a renowned immunologist at Rockefeller University in the United States. It was initially derived from the serum of a 44-year-old Hispanic male HIV patient, whose serum exhibited significant HIV-neutralizing activity. It is one of the most potent broad-spectrum anti-HIV neutralizing antibodies currently available internationally, and the only novel AIDS treatment globally that induces HIV immune recognition. Large-scale production is achieved through gene cloning, transduction, and cell expression purification technologies. Its molecular mechanism of action differs from that of anti-HIV drugs, allowing for synergistic use or combination therapy with other formulations.

[0004] Like other protein drugs, antibodies tend to be unstable at high concentrations. Since the dosage of antibodies is generally large, developing a high-concentration HIV neutralizing antibody formulation would better meet clinical needs and improve patient compliance. Summary of the Invention

[0005] The purpose of this invention is to provide a human immunodeficiency virus neutralizing antibody preparation and its use.

[0006] In a first aspect of the present invention, an antibody liquid formulation is provided, the formulation comprising:

[0007] (a) A therapeutically effective amount of HIV neutralizing antibodies;

[0008] (b) Buffer solutions with a concentration of 10-30 mmol / L;

[0009] (c) stabilizers; and

[0010] (d) Surfactants,

[0011] The concentration of the HIV neutralizing antibody is 90-140 mg / mL.

[0012] The buffer solution is histidine buffer;

[0013] The stabilizer is one or a combination of two or more of sodium chloride, amino acids, and polyols. The amino acid is selected from arginine or histidine, and the polyol is one or a combination of two or more of sucrose, sorbitol, and mannitol. The concentration of sodium chloride is 20-80 mmol / L; the concentration of the amino acid is 10-100 mmol / L; and the concentration of the polyol is 1%-15% by weight, based on the total weight of the liquid formulation.

[0014] The surfactant is Tween 80 or Tween 20, and the concentration of the surfactant is 0.01%-0.08% by weight, based on the total weight of the liquid formulation.

[0015] The pH of the liquid formulation is 5.5-6.5.

[0016] In another preferred embodiment, the heavy chain amino acid sequence of the HIV neutralizing antibody is shown in SEQ ID NO: 1; and the light chain amino acid sequence of the HIV neutralizing antibody is shown in SEQ ID NO: 2.

[0017] In another preferred embodiment, the concentration of the HIV neutralizing antibody is 100-130 mg / mL.

[0018] In another preferred embodiment, the stabilizer is one or a combination of two or more amino acids and polyols.

[0019] In another preferred embodiment, the concentration of sodium chloride is 30-70 mmol / L, more preferably 40-60 mmol / L, and even more preferably 45-55 mmol / L.

[0020] In another preferred embodiment, the concentration of histidine in the histidine buffer is 15-25 mmol / L.

[0021] In another preferred embodiment, the amino acid is arginine, and the concentration of the amino acid is 5-50 mmol / L, more preferably, the concentration of the amino acid is 15-25 mmol / L.

[0022] In another preferred embodiment, the polyol is sucrose, and the concentration of the sucrose is 3%-10% by weight, preferably 4.5%-6.5% by weight.

[0023] In another preferred embodiment, the pH of the liquid formulation is 5.8-6.2.

[0024] In another preferred embodiment, the surfactant is Tween 80, and the concentration of the surfactant is 0.04% to 0.07% by weight.

[0025] In another preferred embodiment, the buffer solution is a histidine buffer solution;

[0026] The amino acid in question is arginine.

[0027] The polyol is sucrose;

[0028] The surfactant is Tween 80.

[0029] In another preferred embodiment, the formulation comprises:

[0030] (a) HIV neutralizing antibodies at a concentration of 100-130 mg / mL;

[0031] (b) 15-25 mmol / L histidine buffer;

[0032] (c) 4.5%-6.5% by weight of sucrose;

[0033] (d) Sodium chloride at a concentration of 45-55 mmol / L;

[0034] (e) 15-25 mmol / L of arginine; and

[0035] (f) 0.04%-0.07% by weight of Tween 80.

[0036] In a second aspect of the invention, a kit is provided, the kit comprising:

[0037] Antibody liquid formulations as described in the first aspect of the present invention; and

[0038] Containers used to hold the liquid formulation.

[0039] In another preferred embodiment, the kit also includes instructions for use.

[0040] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0041] Through extensive and in-depth research, the inventors discovered that a liquid formulation composed of a high concentration of human immunodeficiency virus neutralizing antibody (HIV neutralizing antibody) and a specific concentration of histidine-containing buffer system, stabilizers (such as sucrose, arginine, and sodium chloride), and surfactants (such as Tween) maintains excellent stability in terms of appearance, protein concentration, turbidity, and purity under pH conditions of 5.5-6.5. Furthermore, the chemical stability of the formulation is significantly improved, the rate of charge isomer formation in the formulation is significantly reduced, and the shelf life of the formulation is effectively extended. Based on this, the present invention was completed.

[0042] As used herein, "liquid formulation" means a preparation in a form that allows the biological activity of the active ingredient to be effective and contains no other components that would have unacceptable toxicity to a subject administering the formulation. Subjects include mammals, preferably humans.

[0043] As used herein, "antibody stability" refers to the ability of an antibody to substantially retain its physical and / or chemical stability and / or biological activity after storage. Storage period is generally selected based on the intended shelf life of the formulation. Various analytical techniques for measuring antibody stability are well known in the art.

[0044] Stability can be determined at a selected temperature and for a selected time. Preferably, the formulation is stable for at least one month at room temperature or 30°C to 40°C and / or stable for at least two years at approximately 2-8°C.

[0045] If the antibody in the formulation shows essentially no signs of aggregation, precipitation and / or denaturation when examined by the naked eye for color and / or transparency, or by UV light scattering or size exclusion chromatography, the antibody maintains its physical stability.

[0046] If an antibody is considered to retain its biological activity as defined below at a specific time point due to its chemical stability, then the antibody maintains its chemical stability in the formulation. Chemical stability can be assessed by detecting and quantifying chemical changes in the antibody. Chemical changes may involve size changes (e.g., shearing), which can be assessed, for example, using size exclusion chromatography, SDS-PAGE, and / or matrix-assisted laser desorption / ionization / time-of-flight mass spectrometry (MALDI / TOF MS). Other types of chemical changes include charge changes (e.g., changes due to deamidation), which can be assessed, for example, by ion exchange chromatography.

[0047] If an antibody in a formulation has the biological activity intended for its use, then the antibody retains its biological activity in the formulation. For example, if the biological activity of the antibody in the formulation is within approximately 70% to 130% (within the margin of error of the assay) of the biological activity exhibited when the formulation was prepared, it is considered to have retained its biological activity (e.g., determined by an antigen binding assay).

[0048] As used herein, "therapeutic effective amount" or "effective amount" means, in a pharmacological sense, within the scope of this invention, an effective amount for the prevention or treatment of disease. The antibody is effective for the treatment of the disease. "Treatment" refers to both therapeutic treatment and preventative or defensive measures. Subjects requiring treatment include those already suffering from the relevant condition, as well as those requiring prevention of the relevant disease.

[0049] As used herein, “human immunodeficiency virus neutralizing antibody of the present invention”, “human immunodeficiency virus neutralizing antibody”, “HIV neutralizing antibody of the present invention” and “HIV neutralizing antibody” can be used interchangeably and all refer to antibodies containing the heavy chain and light chain sequences shown in SEQ ID NO: 1 and 2.

[0050] The heavy and light chain sequences of the HIV neutralizing antibody in this invention are as follows:

[0051] Heavy chain (SEQ ID NO: 1):

[0052] QVQLLQSGAAVTKPGASVRVSCEASGYNIRDYFIHWWRQAPGQGLQWVGWINPKTGQPNNPRQFQGRVSLTRHASWDFDTFSFYMDLKALRSDDTAVYFCARQRSDYWDFDVW GSGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0053] Light chain (SEQ ID NO: 2):

[0054] DIQMTQSPSSSLSASVGDTVTITCQANGYLNWYQQRRGKAPKLLIYDGSKLERGVPSRFSGRRWGQEYNLTINNLQPEDIATYFCQVYEFVVPGTRLDLKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0055] Antibody liquid preparation

[0056] The antibody liquid formulation of the present invention mainly comprises:

[0057] (a) A therapeutically effective amount of HIV neutralizing antibodies;

[0058] (b) Buffer solutions with a concentration of 10-30 mmol / L;

[0059] (c) Stabilizers;

[0060] (d) Surfactants.

[0061] The therapeutically effective amount of the antibody present in the formulation of this invention is determined by taking into account the required dose volume and administration method. In this invention, the antibody concentration is 90-150 mg / mL, preferably 100-130 mg / mL. This invention includes a range using any combination of the above values ​​as the upper and / or lower limits.

[0062] The buffer system used in the formulation of the present invention is a buffer system containing histidine. Histidine may exist alone in this buffer system, or in the following forms: histidine hydrochloride, histidine acetate, histidine phosphate, histidine sulfate, etc. In a preferred embodiment, histidine in the buffer system exists alone or in the form of histidine hydrochloride.

[0063] The stabilizer in the formulation of this invention can be selected from one or a combination of two or more of sodium chloride, amino acids, and polyols. The amino acid is selected from arginine or histidine. Preferably, the amino acid is arginine, and its concentration is 5-50 mmol / L, more preferably 15-25 mmol / L.

[0064] The osmotic pressure regulator of this invention is mainly composed of a polyol and sodium chloride, wherein a "polyol" is a substance having multiple hydroxyl groups and includes sugars (reducing sugars and non-reducing sugars), sugar alcohols, and sugar acids. A "reducing sugar" is a sugar containing a hemiacetal group, capable of reducing metal ions or covalently reacting with lysine and other amino groups in proteins, while a "non-reducing sugar" is a sugar that does not possess the aforementioned characteristics of a reducing sugar. Examples of reducing sugars include fructose, mannose, maltose, lactose, arabinose, and xylose. Non-reducing sugars include sucrose, trehalose, sorbitol, mesotriose, and raffinose. Examples of sugar alcohols include mannitol, xylitol, erythritol, threitol, sorbitol, and glycerol. As for sugar acids, they include L-gluconic acid and its metal salts. If the formulation is required to be freeze-thaw stable, the polyol is preferably one that does not crystallize at freezing temperatures (e.g., -20°C) so that it destabilizes the antibodies in the formulation.

[0065] The amount of polyol used can be varied according to the required isotonicity of the formulation. The formulation of the present invention is preferably isotonic. The amount of polyol added can also be varied according to the molecular weight of the polyol. The preferred polyol of the present invention is a sugar alcohol. In a preferred embodiment of the present invention, the polyol is sucrose, and the concentration of sucrose is 1%-15% by weight, more preferably 3%-10% by weight, and even more preferably 4.5%-6.5% by weight. The present invention includes a range using any combination of the above values ​​as the upper and / or lower limits.

[0066] The concentration of sodium chloride is 30-70 mmol / L, preferably 40-60 mmol / L, and more preferably 45-55 mmol / L.

[0067] The surfactant in the formulations of this invention is preferably a nonionic surfactant, such as sorbitan polyoxyethylene ether fatty acid esters (e.g., sorbitan polyoxyethylene (20) ether laurate, sorbitan polyoxyethylene (20) ether oleate, etc.) or poloxamer (e.g., poloxamer 188). The amount of surfactant added is such that it reduces antibody aggregation in the formulation and / or reduces particle formation and / or adsorption. A preferred surfactant in this invention is Tween, such as Tween 80. In a preferred embodiment, the concentration of Tween is 0.01 wt% to 0.08 wt%, preferably 0.04 wt% to 0.07 wt%. This invention includes a range of values ​​using any combination of the above values ​​as upper and / or lower limits.

[0068] This invention uses a buffer system to adjust the pH of the formulation to control the pH within the range of 5.5-6.5. In some embodiments, the pH of the formulation is between 5.5 and 6.4, 5.6 and 6.3, 5.7 and 6.2, 5.8 and 6.1, or 5.9 and 6.0. This invention includes using any combination of the above values ​​as the upper and / or lower limits. In some preferred embodiments, the pH of the formulation is 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, or 6.5.

[0069] It should be understood that, in addition to histidine, the buffer system of the present invention may further include one or more other buffering components, which, in combination with other buffering components, control the pH of the formulation within the aforementioned range. Suitable other buffering components include citrates, phosphates, acetates (e.g., sodium acetate), succinates (e.g., sodium succinate), etc.

[0070] In a preferred embodiment of the present invention, the buffer system is a histidine buffer system, wherein the histidine concentration is 10-30 mmol / L, preferably 15-25 mmol / L. The present invention includes a range of values ​​using any combination of the above values ​​as the upper and / or lower limits. In another embodiment, the pH of the formulation is adjusted using an inorganic acid such as citric acid, acetic acid, or phosphoric acid.

[0071] Through extensive experiments and data screening, the inventors discovered that when the concentration of histidine in the formulation of this invention is below 15 mg / ml, the buffering capacity of the buffer system will be significantly limited. When the concentration of histidine is above 25 mg / ml, it does not improve the stability of the formulation and may cause adverse reactions.

[0072] In addition, the inventors conducted repeated experiments and screened a large number of components and contents of various buffer systems, and finally obtained the histidine system of the present invention from a large number of buffer systems. They also found that the stability of antibody proteins was significantly improved in this system.

[0073] The formulations of this invention may include one or more other pharmaceutically acceptable carriers, excipients, or stabilizers, such as those described in Remington's Pharmaceutical Sciences 16th edition, Osol, A.Ed. (1980), provided they do not adversely affect the desired characteristics of the formulation. Acceptable carriers, excipients, or stabilizers are non-toxic to the recipient at the dosage and concentration used, and include other cosolvents; antioxidants, including ascorbic acid and methionine; integrators, such as EDTA; metal complexes (e.g., Zn-protein complexes); biodegradable polymers, such as polyesters; and / or salt-forming anti-ion.

[0074] The formulation of the present invention can be prepared by combining various components at certain concentrations using methods known in the art.

[0075] One type of optimization method mainly includes the following steps:

[0076] The HIV neutralizing antibody of this invention was concentrated by centrifugation (4500 rpm, 4-10°C) using Ultracel-30K ultrafiltration centrifuge tubes and then transferred to different formulation buffers. The protein concentration was adjusted to the desired level using the formulation buffers. The formulation was then sterilized by filtration using a 0.22 μm Millex syringe filter. The prepared formulation was packaged for ease of use; the packaging material could be a glass bottle, a pre-filled syringe, or a pen syringe.

[0077] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0078] Compared with the prior art, the present invention has the following beneficial effects:

[0079] 1. The formulation of the present invention can effectively reduce the chemical degradation rate and aggregate formation of the HIV neutralizing antibody of the present invention, improve the physicochemical stability of the antibody, and extend the shelf life of the product.

[0080] 2. By controlling the pH value of the formulation of the present invention and removing potentially adverse reaction-causing components (such as citric acid), adverse reactions at the injection site can be eliminated or reduced, thereby improving patient comfort.

[0081] The present invention will be further described below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions as described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0082] Size exclusion chromatography (SEC-HPLC) was used to determine the purity, polymer content, and degradation product content of the HIV neutralizing antibody of this invention. This method was performed using a Tosoh Bioscience TSK-Gel 3000SW. XLAn SEC column was used as the stationary phase. A mobile phase consisting of 100 mM sodium phosphate, 50 mM sodium chloride, 10% acetonitrile, and pH 7.0 was prepared. The test sample and HIV neutralizing antibody reference were prepared by diluting with formulation buffer to a target protein concentration of 1 mg / mL. After injection of a series of known gel-filtered standards (670 kDa–1.35 kDa), the test sample and reference were injected into the column. The peak areas and elution times of the reference and test samples were determined. Purity results were reported as a percentage of the HIV neutralizing antibody monomer peak relative to all other peaks (i.e., high and low molecular weight substances).

[0083] Whole-column imaging cIEF (cIEF) was used to determine the charge isomeric properties of the HIV neutralizing antibody of this invention. Samples and standards were prepared in a carrier ampholyte at pH 8–10.5 using 0.35% methylcellulose and 10% glycerol additives. Quantification was performed using pI markers pI 7.55 and pI 10.10. The sample, HIV neutralizing antibody reference, and pI markers were injected into a fluorocarbon-coated fused silica capillary. Focusing was performed at 1.5 kV for 1 minute and at 3.0 kV for 12 minutes. The focused protein region was photographed using a charge-coupled device (CCD) camera detectable at 280 nm. The chromatographic peaks of the HIV neutralizing antibody subtypes were integrated, and the pI values ​​were quantified based on their pixel positions relative to the internal pI markers.

[0084] Example 1 Formulation Preparation

[0085] (1) Prepare the following two buffer solutions:

[0086] Buffer 1: 20mM histidine, 50mM sodium chloride, 20mM arginine, 5% sucrose, pH 6.0

[0087] Buffer 2: 20mM histidine, 50mM sodium chloride, 20mM arginine, 5% sucrose, 10% Tween 80, pH 6.0

[0088] (2) The buffer solution of the intermediate HIV neutralizing antibody to be replaced with buffer 1 was replaced by dialysis, and the concentration of HIV neutralizing antibody was controlled at about 100 mg / mL to obtain the intermediate HIV neutralizing antibody after the buffer solution was replaced.

[0089] (3) Add buffer 2 at a ratio of 5 g / kg of intermediate to obtain the formulated active pharmaceutical ingredient. The ingredients are 20 mM histidine, 50 mM sodium chloride, 20 mM arginine, 5% sucrose, 0.05% Tween 80, pH 6.0. The concentration of HIV neutralizing antibody is approximately 100 mg / mL.

[0090] Example 2: Comparison of Formulations

[0091] For the following two prescriptions, drug stability assessments were conducted at different concentrations.

[0092] Formulation A: 8.06mM sodium phosphate, 1.47mM potassium phosphate, 136.9mM sodium chloride, 2.68mM potassium chloride, Tween 80, pH 7.0.

[0093] Formulation B: 20mM histidine, 50mM sodium chloride, 20mM arginine, 5% sucrose, Tween 80, pH 6.0.

[0094] As shown in Table 1, based on different Tween 80 values, formulation A corresponds to formulations 1, 2, 3, and 4; formulation B corresponds to formulations 5 and 6.

[0095] Table 1. Pharmaceutical Ingredients

[0096]

[0097] (1) Solution appearance: After being stored at high temperature or for a long time, the appearance of the preparation may change negatively, such as darkening of color or turbidity.

[0098] After the formulation was prepared, an appearance test was performed. As shown in Table 2, formulation B exhibited good clarity and solubility at high concentrations (>70 mg / mL).

[0099] Table 2. Appearance Inspection of Formulations

[0100]

[0101] Meanwhile, as shown in Table 3, the appearance of formulation B was relatively stable at both 5℃ and 40℃.

[0102] Table 3. Appearance detection of formulations at different temperatures

[0103]

[0104] (2) Particle size: After being stored at high temperature or for a long time, the particle size of drug molecules in the preparation tends to increase.

[0105] As shown in Table 4, at 5℃ and 40℃, the particle size of formulation A at 70 mg / mL changed significantly over time, while formulation B maintained relatively stable particle size at higher concentrations. In repeated freeze-thaw experiments, the particle sizes of formulations 5 and 6 were basically the same before and after freeze-thaw, while the particle size of formulation 2 increased significantly.

[0106] Table 4. Particle size detection of formulations at different temperatures

[0107]

[0108] (3) Purity: After being stored at high temperature or for a long time, the purity of the antibody tends to decrease, and the content of high polymer (HMW) tends to increase.

[0109] As shown in Table 5, although the purity of each formulation did not show a trend of change within 8 weeks at 5℃, the purity of formulation A decreased sharply after 8 weeks at 40℃, with an increase in high molecular weight impurities. Especially when the concentration was increased to 70 mg / mL, the acidic component increased by 187%. Meanwhile, the purity of formulation B changed slowly at a concentration of 100 mg / mL, with the acidic component increasing by only 87%.

[0110] Table 5. Purity detection of formulations at different temperatures

[0111]

[0112] (4) Charge isomers: After being stored at high temperature or for a long time, the acidic components of the antibody tend to increase and the main peak decreases.

[0113] As shown in Table 6, although the charge isomer composition of each formulation did not change significantly at 5°C, formulations 1 and 2 showed obvious trend changes after 8 weeks at 40°C, with their main peak components decreasing by 73% and 75%, respectively. In contrast, the charge isomer changes of formulations 5 and 6 were relatively insignificant, with their main peak components decreasing by only 55% and 49%, respectively.

[0114] Table 6. Detection of charge isomers of formulations at different temperatures

[0115]

[0116] In summary, when the concentration is increased to 70 mg / mL, formulation A exhibits poor stability in terms of particle size, purity, and charge isomer composition. Meanwhile, formulation B shows significantly better stability than formulation A at concentrations of 100 mg / mL or higher.

[0117] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims. sequence list <110> Frontier Biopharmaceuticals (Nanjing) Co., Ltd. <120> A human immunodeficiency virus neutralizing antibody preparation and its uses <130> P2021-1583 <160> 2 <170> SIPOSequenceListing 1.0 <210> 1 <211> 453 <212> PRT <213> Artificial Sequence <400> 1 Gln Val Gln Leu Leu Gln Ser Gly Ala Ala Val Thr Lys Pro Gly Ala 1 5 10 15 Ser Val Arg Val Ser Cys Glu Ala Ser Gly Tyr Asn Ile Arg Asp Tyr 20 25 30 Phe Ile His Trp Trp Arg Gln Ala Pro Gly Gln Gly Leu Gln Trp Val 35 40 45 Gly Trp Ile Asn Pro Lys Thr Gly Gln Pro Asn Asn Pro Arg Gln Phe 50 55 60 Gln Gly Arg Val Ser Leu Thr Arg His Ala Ser Trp Asp Phe Asp Thr 65 70 75 80 Phe Ser Phe Tyr Met Asp Leu Lys Ala Leu Arg Ser Asp Asp Thr Ala 85 90 95 Val Tyr Phe Cys Ala Arg Gln Arg Ser Asp Tyr Trp Asp Phe Asp Val 100 105 110 Trp Gly Ser Gly Thr Gln Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 435 440 445 Leu Ser Pro Gly Lys 450 <210> 2 <211> 206 <212> PRT <213> Artificial sequence <400> 2 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Thr Val Thr Ile Thr Cys Gln Ala Asn Gly Tyr Leu Asn Trp Tyr 20 25 30 Gln Gln Arg Arg Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp Gly Ser 35 40 45 Lys Leu Glu Arg Gly Val Pro Ser Arg Phe Ser Gly Arg Arg Trp Gly 50 55 60 Gln Glu Tyr Asn Leu Thr Ile Asn Asn Leu Gln Pro Glu Asp Ile Ala 65 70 75 80 Thr Tyr Phe Cys Gln Val Tyr Glu Phe Val Val Pro Gly Thr Arg Leu 85 90 95 Asp Leu Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro 100 105 110 Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu 115 120 125 Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn 130 135 140 Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser 145 150 155 160 Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala 165 170 175 Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly 180 185 190 Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 195 200 205

Claims

1. An antibody liquid formulation, characterized by, The formulation consists of the following components: (a) HIV neutralizing antibodies at a concentration of 100-130 mg / mL; (b) 20 mmol / L histidine buffer; (c) 5% by weight of sucrose; (d) 50 mmol / L sodium chloride; (e) 20 mmol / L arginine; and (f) 0.05% - 0.065% by weight of Tween 80; Furthermore, the pH of the liquid formulation is 5.5-6.5; the buffer solution is water. The heavy chain amino acid sequence of the HIV neutralizing antibody is shown in SEQ ID NO: 1; the light chain amino acid sequence of the HIV neutralizing antibody is shown in SEQ ID NO:

2.

2. The liquid formulation of claim 1, wherein, The formulation consists of the following components: (a) 100 mg / mL HIV neutralizing antibody; (b) 20 mmol / L histidine buffer; (c) 5% by weight of sucrose; (d) 50 mmol / L sodium chloride; (e) 20 mmol / L arginine; and (f) 0.05% - 0.065% by weight of Tween 80; The liquid preparation has a pH of 6; the buffer solution is a medium of water.

3. The liquid formulation of claim 1, wherein The formulation consists of the following components: (a) 100 mg / mL HIV neutralizing antibody; (b) 20 mmol / L histidine buffer; (c) 5% by weight of sucrose; (d) 50 mmol / L sodium chloride; (e) 20 mmol / L arginine; and (f) 0.05% by weight of Tween 80; The liquid preparation has a pH of 6; the buffer solution is a medium of water.

4. The liquid formulation of claim 1, wherein, The formulation consists of the following components: (a) HIV neutralizing antibody at a concentration of 130 mg / mL; (b) 20 mmol / L histidine buffer; (c) 5% by weight of sucrose; (d) 50 mmol / L sodium chloride; (e) 20 mmol / L arginine; and (f) 0.065% by weight of Tween 80; The liquid preparation has a pH of 6; the buffer solution is a medium of water.

5. A kit characterized in that, The kit comprises: an antibody liquid formulation as described in any one of claims 1-4; and Containers used to hold the liquid formulation.

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