CN116712390B - High-concentration high-stability antibody preparation and preparation method thereof - Google Patents

High-concentration high-stability antibody preparation and preparation method thereof Download PDF

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CN116712390B
CN116712390B CN202310976752.0A CN202310976752A CN116712390B CN 116712390 B CN116712390 B CN 116712390B CN 202310976752 A CN202310976752 A CN 202310976752A CN 116712390 B CN116712390 B CN 116712390B
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antibody
preparation
histidine
concentration
buffer
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CN116712390A (en
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庄超
黄峥
高煜
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Shanghai Lanyi Pharmaceutical Technology Co ltd
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    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
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    • C07—ORGANIC CHEMISTRY
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    • C07K2317/00—Immunoglobulins specific features
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    • C07K2317/00—Immunoglobulins specific features
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    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
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    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides a high-concentration high-stability antibody preparation and a preparation method thereof, belonging to the field of antibody preparations, wherein the antibody preparation comprises an anti-C5 antibody, a light chain sequence is SEQ ID NO.1, and a heavy chain sequence is SEQ ID NO.2 or a long-acting modified sequence thereof; also included is a buffer system: 20-50mM histidine-histidine hydrochloride, 70-110mM arginine, 25-50mM sodium chloride and 0.03% PS80. The concentration of the conventional antibody preparation is generally below 100mg/mL, and the concentration of the antibody in the buffer solution system reaches about 200mg/mL. And the antibody preparation of the invention shows good long-term stability, affinity and bioavailability.

Description

High-concentration high-stability antibody preparation and preparation method thereof
Technical Field
The invention belongs to the field of antibody preparations, and particularly relates to a high-concentration high-stability antibody preparation and a preparation method thereof.
Background
With the development of bioantibody drug therapies, high concentrations of the active ingredient (about 20mg/mL to about 100-150mg/mL, even 200mg/mL or higher) are required for intravenous or subcutaneous high concentration administration. In the field of treatment of chronic diseases with a large single dose, such as PNH (paroxysmal sleep hemoglobinuria, which is manifested by chronic intravascular hemolysis), in order to achieve autonomous administration of a patient, to improve convenience and therapeutic compliance of administration of the patient, an antibody preparation with a small administration volume and high concentration needs to be developed. However, antibody drugs can have problems such as increased viscosity, protein aggregation, and altered charge heterogeneity during concentration at high concentrations, which can lead to decreased activity of the active antibody, increased immunogenicity, and the like. Thus, there is a need to improve the formulation of antibody drugs to increase antibody solubility, decrease viscosity, and address the problems of protein aggregation, charge heterogeneity changes.
The improvements to antibody pharmaceutical formulations mainly include the following:
I. antibodies in the formulation;
calcium and acetate or buffer:
the viscosity of the antibody formulation is reduced by adding calcium salts and/or acetate salts (and/or buffers) to the formulation: determination methods the viscosity of the protein formulation is determined before and after addition of calcium salt and/or acetate (and/or buffer);
excipient in formulation:
the excipient can impart or enhance the stability of the protein preparation and influence the efficacy and immunogenicity of the drug to some extent, so that development of the protein preparation of the excipient with stability and safety is required. The excipient may be a buffer, a stabilizer, a surfactant, or the like.
EA5 mab is a humanized monoclonal antibody that specifically targets complement C5. Complement C5 can be converted to C5a and C5b after cleavage, where C5a is an important pro-inflammatory molecule and C5b can continue to form C5b-9, the tapping complex (MAC) that leads to cell perforation or lysis. Deposition of the tapping complex on the surface of erythrocytes in PNH patients can cause erythrocyte lysis, the major clinical manifestation of PNH. EA5 mab blocks C5 cleavage and prevents the formation of C5a and C5b-9 and is therefore useful for the effective treatment of PNH. In addition, the composition has good therapeutic effects on diseases caused by complement end activation, such as aHUS, gMG and the like.
PNH (paroxysmal sleep hemoglobinuria) is a rare and life-threatening hematopoietic stem cell clonal disease with prevalence of about thirteen parts per million, and is clinically manifested mainly by hemolytic anemia, hemoglobinuria, venous thrombosis, bone marrow hematopoietic failure, smooth muscle dysfunction, and the like. Intravascular hemolysis caused by the complement system attacking erythrocytes is a central factor in PNH morbidity and mortality. Ext> CDext> 59ext> isext> theext> mostext> importantext> complementext> complexext> (ext> MACext>)ext> inhibitorext> onext> theext> surfaceext> ofext> erythrocytesext>,ext> andext> PNHext> patientsext> undergoext> acquiredext> somaticext> mutationext> dueext> toext> phosphatidylinositolext> glycanext> (ext> PIGext> -ext> aext>)ext> geneext> locatedext> onext> theext> Xext> chromosomeext> (ext> xpext> 22.1ext>)ext>,ext> resultingext> inext> failureext> ofext> CDext> 59ext> toext> bindext> toext> theext> cellext> membraneext> ofext> erythrocytesext> dueext> toext> theext> defectext> ofext> glycosylext> phosphatidylinositolext> (ext> gpiext>)ext> synthesisext>,ext> andext> eventuallyext> lossext> ofext> functionext> inext> inhibitingext> activationext> ofext> theext> complementext> pathwayext>.ext> The main treatment methods of PNH include complement inhibitors, glucocorticoids, allogeneic hematopoietic stem cell transplantation, etc., wherein complement C5 inhibitors Soliris (Eculizumab) or Ultomiris (Ravulizumab) are standard treatment of PNH, can effectively control intravascular hemolysis, reduce thrombosis and improve long-term survival rate, but the medicines are expensive and need intravenous injection, and patients need regular hospital admission and infusion, thus bringing great inconvenience to life. Therefore, the development of a safe and effective subcutaneous complement C5 inhibitor for treatment of PNH patients is of great significance.
PNH is a disease caused by typical complement activation, EA5 antibody has great therapeutic potential for other diseases caused by complement activation as well, such as rheumatoid arthritis, antiphospholipid antibody syndrome, lupus nephritis, ischemia reperfusion injury, atypical hemolytic uremic syndrome (aHUS), typical hemolytic uremic syndrome, paroxysmal Nocturnal Hemoglobinuria (PNH), compact deposition disease, neuromyelitis optica, multifocal motor neuropathy, multiple sclerosis, macular degeneration, HELLP syndrome, spontaneous abortion, thrombotic thrombocytopenic purpura, oligoimmunovasculitis, epidermolysis bullosa, recurrent abortion, traumatic brain injury, myocarditis, cerebrovascular disorders, peripheral vascular disorders, renal vascular disorders, mesenteric/intestinal vascular disorders, vasculitis, allergic purpura nephritis, systemic lupus erythematosus-associated vasculitis, rheumatoid arthritis-associated vasculitis, immunocomplex vasculitis, high-altitude disease, dilated cardiomyopathy, diabetes mellitus blood vessel, sichuan disease, venous embolism, stent placement, rotational atherectomy, restenosis following percutaneous transluminal coronary angioplasty, myasthenia gravis, collectin's disease, dermatomyositis, paroxysmal cold hemoglobinuria, antiphospholipid syndrome, graves' disease, atherosclerosis, alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, graft rejection, hashimoto's thyroiditis, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, godpasture's syndrome, digoss' disease, and catastrophic antiphospholipid syndrome.
Disclosure of Invention
In order to solve the problems, the invention provides an anti-C5 antibody and a preparation thereof, optimizes the molecular design of the antibody and a buffer system of the preparation, and aims to improve the overall stability of the antibody preparation.
In the present invention, "PS80" refers to polysorbate-80, also known as Tween-80.
In the present invention, "histidine-histidine hydrochloride" refers to histidine sodium histidine hydrochloride+histidine hydrochloride buffer.
In one aspect, the invention provides an anti-C5 antibody.
The anti-C5 antibody is an engineered antibody, the light chain sequence is SEQ ID NO.1, and the heavy chain sequence is SEQ ID NO.2 or a long-acting engineered sequence thereof. The antibody molecule improves the solubility and stability of the antibody by engineering an Fc region; raising the isoelectric point of the antibody to avoid loss of antibody in downstream processes; while increasing the affinity of the antibody for FcRn to achieve a longer in vivo half-life.
Preferably, the long-acting modification sequence of the heavy chain sequence is SEQ ID NO.3.
In another aspect, the invention provides an antibody preparation comprising the foregoing anti-C5 antibody.
The content of the anti-C5 antibody is 0-200mg/L, preferably 160-200mg/mL, and more preferably 180mg/mL.
The antibody preparation also comprises a buffer system.
The buffer system comprises: 20-50mM histidine-histidine hydrochloride, 70-110mM arginine, 25-50mM sodium chloride and 0.03% PS80.
The histidine-histidine hydrochloride content may be selected from: 20-30mM, 30-40mM, 40-50mM, 25-35mM, 25-45mM.
The arginine may be present in an amount selected from: 70-80mM, 80-90mM, 90-100mM, 100-110mM, 70-90mM, 80-100mM, 90-110mM, 100-110mM.
The sodium chloride content may be selected from: 25-30mM, 25-35mM, 30-50mM, 40-50mM, 35-40mM, 25-45mM, 30-45mM.
Preferably, the buffer system comprises: 20mM histidine-histidine hydrochloride, 70-110mM arginine, 25-50mM sodium chloride and 0.03% PS80.
More preferably, the buffer system comprises: 20mM histidine-histidine hydrochloride, 80mM arginine, 25-50mM sodium chloride and 0.03% PS80.
Further preferably, the buffer system comprises: 20mM histidine-histidine hydrochloride, 80mM arginine, 50mM sodium chloride and 0.03% PS80.
The pH of the buffer system is 6.0-6.5.
Preferably, the pH of the buffer system is between 6.0 and 6.3.
Further preferably, the buffer system has a pH of 6.3.
The invention also provides buffer systems for use in antibody formulations.
In a further aspect, the invention provides the use of an anti-C5 antibody or antibody preparation as described above in the manufacture of a medicament.
The pharmaceutical indications are selected from: rheumatoid arthritis, antiphospholipid antibody syndrome, lupus nephritis, ischemia reperfusion injury, atypical hemolytic uremic syndrome (aHUS), atypical hemolytic uremic syndrome, PNH, dense matter deposition disease, neuromyelitis optica, multifocal motor neuropathy, multiple sclerosis, macular degeneration, HELLP syndrome, spontaneous abortion, thrombotic thrombocytopenic purpura, oligoimmunovasculitis, epidermolysis bullosa, recurrent abortion, traumatic brain injury, myocarditis, cerebrovascular disorders, peripheral vascular disorders, renal vascular disorders, mesenteric/intestinal vascular disorders, vasculitis, allergic purpura nephritis, systemic lupus erythematosus-associated vasculitis, rheumatoid arthritis-associated vasculitis, immune complex vasculitis, high-amp disease, dilated cardiomyopathy, diabetic vasculopathy, kawasaki disease, venous air embolism, stent placement, rotational atherectomy, restenosis following percutaneous transluminal coronary angioplasty, myasthenia gravis, collectin's disease, dermatomyositis, paroxysmal cold hemoglobinuria, antiphospholipid syndrome, graves' disease, atherosclerosis, alzheimer's disease, systemic inflammatory response sepsis, septic shock, spinal cord injury, glomerulonephritis, graft rejection, hashimoto's thyroiditis, type I diabetes, psoriasis, pemphigus, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, godpassch's syndrome, digoss' disease, or catastrophic antiphospholipid syndrome.
Preferably, the indication of the medicament is PNH.
In yet another aspect, the invention provides a medicament comprising the foregoing anti-C5 antibody or antibody preparation.
Preferably the medicament is for use in the treatment of PNH.
Preferably, the medicament is in the form of injection.
In yet another aspect, the invention provides the use of the buffer system described above in the preservation of antibody drugs.
Preferably, the antibody drug is an anti-C5 antibody-related drug, such as the preservation of anti-C5 antibodies described above.
The invention has the beneficial effects that:
compared with the prior art, the invention improves the stability of the anti-C5 antibody, improves the related buffer system of the antibody preparation, solves the problems of high viscosity, aggregation, change of charge heterogeneity and the like of the antibody preparation, and improves the stability of the corresponding high-concentration liquid preparation of the antibody. The concentration of the conventional antibody preparation is generally below 100mg/mL, and the concentration of the antibody in the buffer solution system reaches about 200mg/mL. And the antibody preparation of the invention shows good long-term stability, affinity and bioavailability.
Drawings
FIG. 1 shows the inhibitory activity of various concentrations of EA5 antibody on Complement Dependent Cytotoxicity (CDC).
FIG. 2 is a graph of ICR mice given a single intravenous and subcutaneous injection of 30mg/kg of EA5 antibody.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are not intended to limit the present invention, but are merely illustrative of the present invention. The experimental methods used in the following examples are not specifically described, but the experimental methods in which specific conditions are not specified in the examples are generally carried out under conventional conditions, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.
Example 1 preparation of an anti-C5 antibody preparation
(1) Preparation of antibodies
A highly stable anti-C5 antibody having the heavy chain sequence of SEQ ID No.2: the light chain sequence is SEQ ID NO.1. Long-acting modification is carried out on the heavy chain of the modified antibody to increase half-life, and the heavy chain sequence of the modified antibody is SEQ ID NO.3; the light chain sequence is SEQ ID NO.1.
The preparation method of the antibody of the embodiment is a conventional preparation method in the field, an expression vector is constructed, a high-expression stable cell strain is constructed by transferring into a host cell CHO in a stable rotation mode, batch feed culture is carried out on seed cells, the anti-C5 antibody is expressed, and after the anti-C5 antibody is obtained after the harvesting, the affinity chromatography and the ion exchange chromatography are carried out.
(2) Preparation of buffer
The buffer is prepared according to a specific formula, and the pH of the buffer is adjusted to 6.3.
(3) Preparation of antibody formulations
The modified antibody of step (1) and the buffer of step (2) were used to prepare a high concentration antibody preparation at a concentration of 180mg/mL antibody. The preparation process is as follows:
setting the rotating speed of a centrifugal machine to 3500 rcf, concentrating and replacing protein into a preparation prescription by using an ultrafiltration centrifugal tube, replacing the protein by 155 times, adjusting the protein concentration to 180mg/mL of target concentration, filtering and subpackaging each prescription sample into a 2R penicillin bottle and a5 mL liquid storage bag in an ultra-clean bench, and capping the penicillin bottle by plugging.
Prescription 1 buffer: 50mM histidine-histidine hydrochloride, 80mM arginine hydrochloride, 25 mM sodium chloride, 0.03% (w/v) PS80. The buffer pH was adjusted to 6.3.
Prescription 2 buffer: 20mM histidine-histidine hydrochloride, 80mM arginine hydrochloride, 25 mM sodium chloride, 0.03% (w/v) PS80.
Prescription 3 buffer: 20mM histidine-histidine hydrochloride, 110mM arginine hydrochloride, 25 mM sodium chloride, 0.03% (w/v) PS80.
Prescription 4 buffer: 20mM citric acid-sodium citrate+70 mM arginine hydrochloride+50 mM sodium chloride+0.03% PS80 pH6.3.
Experimental example 1 quality Property study of antibody preparation
Taking prescription 1 as an example, the appearance, protein concentration, osmolarity, viscosity, SEC-HPLC assay, CEX-HPLC, nrCE-SDS, binding activity, affinity to C5 and CDC effect were determined for the antibody preparation.
(1) Detection method for protein concentrationThe method comprises the following steps: protein content was measured using an ultraviolet spectrophotometer (Shimadzu, model: UV-1900). Extinction coefficient of protein was 1.531 (mg/mL) -1 cm -1 . The micro cuvettes are washed by ultrapure water for 2 to 3 times, ultrapure water is respectively added into the two micro cuvettes, 200 mu L/cuvette is used for blank correction, 200 mu L/cuvette sample is added, and the sample is continuously read for three times. The concentration of the sample was calculated from the extinction coefficient, dilution and OD value.
(2) The osmotic pressure detection method comprises the following steps: the sample is placed on a sample measuring probe of an osmometer, the instrument automatically starts measuring, and the test result is displayed and printed after 1 minute.
(3) The viscosity detection method comprises the following steps: a microVISC pipettes injector is used for sucking a certain volume of sample, the needle of the injector is propped against a sample inlet, and after the sample is loaded, the instrument automatically starts to measure.
(4) Detection column manufacturer of SEC-HPLC: agilent model AdvanceBio SEC 300A, specification 2.7 μm.
High performance liquid HPLC manufacturer: thermo, ultiMate 3000.
The mobile phase composition was 20mM PB+200mM NaCl+100mM Arg,pH6.8.
And integrating by adopting an area normalization method, and calculating a main peak, a polymer peak and a fragment peak after the monomer.
The SEC-HPLC purity detection conditions are specifically as follows:
(5) CEX-HPLC purity
Chromatographic column: manufacturer Sepax, model Proteomix SCX-NP5, specification 4.6X1250 mm; high performance liquid chromatograph HPLC: manufacturer Thermo, model UltiMate 3000. Mobile phase a:20mM MES, pH6.5; mobile phase B:20mM MES+500mM NaCl,pH6.5. The results were quantitatively analyzed by peak area normalization, and the acid component content (sum of all peak area percentages before the main peak), the main component content (main peak area percentage) and the alkaline component content (sum of all peak area percentages after the main peak) were calculated. The details are given in the following table:
CEX-HPLC purity detection method
(6) nrCE-SDS purity
Capillary electrophoresis apparatus: manufacturer SCIEX, model PA800 Plus. Capillary electrophoresis chromatograph: manufacturer G7100A, model aglent. Desalting the sample, adding pH6.5 citric acid-phosphate buffer solution into the mixture, centrifuging to 400 mu L at 13000rpm for 15min, reversely inserting the centrifuged concentrated column into a new centrifuge tube, centrifuging at 6000rpm for 3min, recovering protein, adding 250mM NEM solution, uniformly mixing, heating at 70 ℃ for 10min, cooling, centrifuging at 10000rpm for 3min, and injecting sample. With a PDA detector the detection window is 100 x 800 μm. The sample is introduced from the left end of the capillary, and the effective separation length of the capillary is 20cm. Voltage injection was used for 20 seconds at 5kV. Constant voltage separation is adopted, and the separation voltage is 15kV. The alkaline wash was forward 70psi for 3min, the acidic wash was forward 70psi for 1min, the pure water was forward 70psi for 1min, and the SDS gel buffer was forward 70psi for 10min. And calculating the content of the high molecular component (the sum of all the area percentages of the peaks before the main peak), the content of the main component (the area percentage of the main peak) and the content of the low molecular component (the sum of all the area percentages of the peaks after the main peak) by adopting a peak area normalization method.
(7) Binding activity detection method (ELISA):
250 μg/mL of C5 antigen (purchased from Acro Bio under the trade designation CO5-H52 Ha) was diluted to 0.5 μg/mL with 1 XPBS coating buffer and 100 μl/well coated with 96-well enzyme label overnight at 4deg.C. Plates were washed 4 times, 250 μl/well of blocking solution was added and incubated for 2 hours at room temperature. The plate was washed 4 times, 100. Mu.L/well of the gradient diluted sample was transferred to a 96-well ELISA plate and incubated for 1h at room temperature. Washing the plate for 4 times, adding secondary antibody working solution (diluted 1:20000) into 100 mu L/hole, standing at room temperature, and incubating for 1h. Washing the plate for 4 times, adding TMB color development liquid into 100 mu L/hole, reacting for 10min at room temperature, and adding stop solution. Absorbance at 450nm was read on a microplate reader with 630 nm as a reference wavelength. OD with concentration on the abscissa 450-630nm On the ordinate, do fourParameter curve fitting can directly obtain EC 50 And R is 2 And calculating to obtain the relative binding activity of the sample to be detected. Relative binding Activity = reference (or working reference) EC 50 Sample to be tested EC 50 ×100%。
The main quality attributes are detected as follows:
experimental example 2 affinity detection of antibody preparation
(1) The affinity of the anti-C5 antibody to C5 was detected as follows:
the biotin-labeled C5 was diluted to 5. Mu.g/mL with SD buffer (0.1% BSA,0.01% Tween 20,1 XPBS), the test sample was diluted to 50. Mu.g/mL, triplicate samples were prepared, the diluted test sample, fcRn and SD buffer were added to a black 96-well plate, SA Sensor and black 96-well plate were placed in the instrument, and the program was set for curve fitting and data analysis. The results of the formulation test shown in prescription 1 are as follows:
the results show that the anti-C5 antibodies of the invention have better affinity with C5.
(2) The affinity of the antibody to FcRn was detected as follows:
before the experiment, the pH of SD buffer solution is adjusted to 6.0, the FcRn with biotin mark is diluted to 5 mug/mL by SD buffer solution, then the test sample is diluted to 50 mug/mL, three parts are prepared in parallel, the diluted test sample, fcRn and buffer solution are respectively added into a black 96-well plate, SA Sensor and the black 96-well plate are placed into equipment instrument, and a program is set for curve fitting and data analysis. Eculizumab (see US 6355245, construction of expression plasmid, 293T cells plus PEIUS5736137A were used by transient transfection and buffer was added after one-step affinity chromatography for preservation) as positive control.
The detection results are as follows:
the detection result shows that the binding capacity of the antibody in the antibody preparation provided by the invention to FcRn is superior to that of a positive control.
(3) CDC effects of different concentrations of antibodies were detected. The detection method comprises the following steps:
an appropriate amount of target cells (human Burkitt's lymphoma cells, ATCC CCL-213) was added with 10. Mu.g/mL Rituximab (see U.S. Pat. No.3,182,62A, expression plasmid was constructed, 293t cells were transiently transfected with PEI and used after one-step affinity chromatography with buffer for storage) and incubated for half an hour, antibody drug at a concentration ranging from 216ng/mL to 12500ng/mL was added, incubated for 3 hours, equilibrated to room temperature was removed, cell titer detection reagent (Promega, cat# G7572) was added, and the cells were read after shaking in the dark. Four-parameter curve fitting was performed with antibody drug concentration on the abscissa and RLU value on the ordinate, where RLU value was positively correlated with cell lysis rate. The results are shown in FIG. 1. FIG. 1 shows that as the concentration of antibody becomes lower, the cell lysis rate increases, the complement-induced CDC effect by Rituximab increases, while high concentrations of anti-C5 antibody can inhibit the CDC effect by Rituximab.
Experimental example 3 accelerated stability test
The formulations of the prescribed 1-4 antibodies were left at 40℃for 14 days, and the initial parameters (T0) and 14-day parameters thereof were examined with reference to experimental example 1 to evaluate the stability of the formulations, with the following results:
experimental example 4 subcutaneous injection bioavailability
Taking prescription 1 as an example, the bioavailability of the antibody preparation is studied, and the specific method is as follows: in the experiment, after subcutaneous and intravenous injection of an antibody preparation to a CD-1 mouse, the change of the blood concentration of the antibody in the mouse with time is observed, so that the basic pharmacokinetic parameters of the medicine are obtained, and the basic kinetic characteristics of a test object are known.
The specific method comprises the following steps:
the groups were randomly grouped according to body weight, 6 each, male and female halves. A single tail vein injection or a single subcutaneous neck injection of antibody preparation was performed using ICR mice (8 weeks old, body weight 30 g) at a dose of 10mL/kg.
Sampling time: immediately after the administration (within 1 min), 1 h+ -5 min, 6 h+ -5 min, 24 h+ -5 min, 48 h+ -5 min, 72 h+ -5 min, 96 h+ -5 min, 120 h+ -5 min, 168 h+ -5 min, 240 h+ -5 min, 336 h+ -5 min, 504 h+ -5 min and 672 h+ -5 min; blood collection amount: about 0.2 mL/one/time; collecting animals: 3/time point/gender/group; the blood sampling mode is as follows: the orbital veins. The concentration of antibodies in the plasma samples was analytically detected using methodology-verified ELISA assays. All samples were multiplexed. Concentration and time profiles of antibodies in plasma were analyzed by non-compartmental model.
ELISA method for determining antibody content in blood plasma:
goat anti-human polyclonal antibodies (ex Abcam, cat No. Ab 6858) were coated on hydrophobic elisa plates and EA5 content in plasma was measured using the double antibody sandwich method. The results are shown in fig. 2, and fig. 2 shows: compared with intravenous injection, the preparation is mainly absorbed by lymphatic system after subcutaneous injection, the peak time of the medicine is 24 hours, the average Cmax is 240 mug/mL, and the bioavailability is about 70%.

Claims (9)

1. An antibody preparation, comprising an anti-C5 antibody and a buffer system;
the anti-C5 antibody has a light chain sequence of SEQ ID NO.1 and a heavy chain sequence of SEQ ID NO.3;
the buffer system consists of 20-50mM histidine-histidine hydrochloride, 70-110mM arginine, 25-50mM sodium chloride and 0.03% tween-80;
the content of the anti-C5 antibody is 160-200mg/mL.
2. The antibody formulation of claim 1, wherein the buffer system consists of 20mM histidine-histidine hydrochloride, 70-110mM arginine, 25-50mM sodium chloride, and 0.03% tween-80.
3. The antibody formulation of claim 2, wherein the buffer system consists of 20mM histidine-histidine hydrochloride, 80mM arginine, 25-50mM sodium chloride and 0.03% tween-80.
4. The antibody preparation of claim 3, wherein the buffer system consists of 20mM histidine-histidine hydrochloride, 80mM arginine, 50mM sodium chloride and 0.03% tween-80.
5. The antibody preparation of claim 4, wherein the buffer system has a pH of 6.0-6.5.
6. The antibody preparation of claim 5, wherein the buffer system has a pH of 6.0-6.3.
7. Use of an antibody preparation according to any one of claims 1 to 6 for the manufacture of a medicament for the treatment of one or more of rheumatoid arthritis, atypical hemolytic uremic syndrome, PNH.
8. The use according to claim 7, characterized in that it is a subcutaneous injection of a drug.
9. A subcutaneous injection medicament comprising the antibody formulation of any one of claims 1-6.
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