CN116903712A - Human IgG binding peptide and application thereof - Google Patents

Human IgG binding peptide and application thereof Download PDF

Info

Publication number
CN116903712A
CN116903712A CN202310050588.0A CN202310050588A CN116903712A CN 116903712 A CN116903712 A CN 116903712A CN 202310050588 A CN202310050588 A CN 202310050588A CN 116903712 A CN116903712 A CN 116903712A
Authority
CN
China
Prior art keywords
binding peptide
higg
binding
igg
human igg
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310050588.0A
Other languages
Chinese (zh)
Inventor
金尤美
杨浩
卢晓风
陶泽
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
West China Hospital of Sichuan University
Original Assignee
West China Hospital of Sichuan University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by West China Hospital of Sichuan University filed Critical West China Hospital of Sichuan University
Publication of CN116903712A publication Critical patent/CN116903712A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/305—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F)
    • C07K14/31—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/65—Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Hematology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Urology & Nephrology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a humanized IgG binding peptide and application thereof. The invention obtains a small molecule peptide (humanized IgG binding peptide) capable of showing specific binding to humanized IgG by recognizing Fc segment through screening skeleton protein mutation library. The amino acid sequence is shown in SEQ ID NO. 1. The human IgG binding peptide can be used for loading, purifying, detecting and analyzing therapeutic and diagnostic medicines of human IgG antibodies, and has good application prospect.

Description

Human IgG binding peptide and application thereof
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a humanized IgG binding peptide and application thereof.
Background
Antibodies (imunoglobin G, igG) not only have high affinity and high specificity for antigens, but also exhibit an ultra-long half-life through neonatal Fc receptor (neonatal Fc receptor, fcRn) -mediated recycling, and are ideal therapeutic and diagnostic drugs (Kaplon H, et al, mAbs,2022, 14:2014296). Currently, hundreds of IgG antibodies are available in bulk clinical settings for disease diagnosis and treatment. Over 800 antibodies are in clinical trials for disease diagnosis and treatment.
In addition to being used directly as a drug, antibodies are also widely used as therapeutic or diagnostic drug carriers. For example, some small molecule chemotherapeutic drugs with weak targeting, short half-life and strong toxic and side effects can be coupled with antibodies to prepare Antibody conjugates (ADC) for tumor treatment, so that the curative effect can be greatly improved and the side effects can be reduced. The coupling of contrast agents such as radionuclides and tumor antigen specific antibodies for solid tumor diagnosis can greatly improve the diagnosis accuracy. Some cytokines, such as Interleukin 2 (Interleukin 2), tumor necrosis factor alpha (Tumor necrosis factor alpha, tnfalpha), etc., can strongly activate immune cells to exhibit a superior antitumor effect. However, the receptors for these factors are widely expressed in humans and the direct use of these factors will inevitably activate the immune system in its entirety to produce systemic immunotoxicity. If the factors are fused and expressed with the antibody, the tumor antigen can be targeted and delivered to the tumor site through the specific recognition of the antibody, and the side effect can be reduced while the treatment effect is improved. However, the existing research mainly couples small-molecule chemotherapeutic drugs (or contrast agents) with antibodies through chemical reaction, and has the disadvantages of high technical difficulty, complex process and low success rate. Because of the large molecular weight of antibodies, the structure of antibodies is easily destroyed under extreme coupling conditions, so that the antibodies lose activity. The cytokine therapeutic protein and the antibody are fused and have to be recombined and expressed by genetic engineering, and the method has the defects of high technical difficulty, low yield, high cost and the like. Therefore, if a more simplified method for loading the antibody with the drug can be found, the range of the antibody carrying the drug can be widened, and the application of the antibody targeting drug in disease diagnosis and treatment can be greatly promoted.
Some peptide molecules are known as IgG binding peptides (chok W et al materials,2016,9,994) because they bind IgG by recognizing a specific structure of IgG (such as Fab or Fc). If a small molecule chemotherapeutic drug, a contrast agent or a cytokine and the like are coupled or fused with the IgG binding peptide and then mixed with an antibody, the therapeutic drug or the contrast agent can be hopefully loaded on the antibody through the mediation of the IgG binding peptide for diagnosing and treating diseases. The peptides have the characteristics of good thermal stability, easy high-efficiency recombinant expression by using escherichia coli, low production cost and the like due to small molecular weight. Thus, coupling or fusion expression of therapeutic drugs or contrast agents to these IgG-binding peptides is much easier than coupling or fusion expression directly to antibodies. However, whether this approach can be implemented depends primarily on whether the appropriate IgG-binding peptide is available. The main clinical application is human IgG. However, many IgG-binding peptides reported in the prior art have the disadvantage of not being specific in binding properties. It exhibits binding to IgG from multiple species, and presents great difficulty in preclinical studies related to human IgG. Thus, there is a need to screen for human IgG specific binding peptides, providing more options for drug development.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a human IgG binding peptide and application thereof, and aims to provide a novel IgG binding peptide which has specific binding effect on human IgG, can be used for constructing antibody loading medicines and provides more choices for medicine development.
A human IgG binding peptide has an amino acid sequence shown in SEQ ID NO. 1.
Preferably, it is expressed by the nucleotide sequence shown as SEQ ID NO. 2.
The invention also provides application of the humanized IgG binding peptide in preparation of antibody loading drugs.
Preferably, in the antibody-loaded drug, the active small molecule is loaded on IgG via the humanized IgG-binding peptide.
Preferably, the active small molecule is one or a combination of two or more of a chemotherapeutic drug, a contrast agent and a cytokine.
Preferably, the human IgG-binding peptide and the small active molecule are bound by means of coupling or fusion expression.
The invention also provides an antibody loading drug which is obtained by loading the active small molecules on IgG through the humanized IgG binding peptide.
Preferably, the active small molecule is one or a combination of two or more of a chemotherapeutic drug, a contrast agent and a cytokine.
Preferably, the human IgG-binding peptide and the small active molecule are bound by means of coupling or fusion expression.
The invention also provides the application of the humanized IgG binding peptide in purification or detection of IgG.
In the present invention, the "antibody-loaded drug" refers to a pharmaceutical preparation prepared by using antibody IgG as a carrier. By "active small molecule" is meant a molecule having a specific biological, chemical or physical function in an organism, such as: chemotherapy drugs, contrast agents or cytokines, etc.
The invention obtains a small molecular peptide (human IgG binding peptide) capable of showing specific binding to human IgG by recognizing Fc segment through screening skeleton protein mutation library, and the small molecular peptide can be used for drug loading, purification and detection analysis of human IgG antibody, and has good application prospect.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 is a protein backbone mutation library construction strategy (-: identical amino acids, X: mutant amino acids);
FIG. 2 shows the results of screening of hIgG binding peptide phage display library and (A) phage library capacity assay. (B) The number of hIgG binding peptide-displaying phages screened in rounds 1-5 (R1-R5) was varied.
FIG. 3 is an analysis of hIgG binding characteristics of round 5 phage clones;
FIG. 4 is an amino acid sequence analysis of round 5 of phage clones;
FIG. 5 shows the binding site and specificity analysis of ZHX display phage to hIgG;
FIG. 6 shows the recombinant expression, purification and identification results of Zhx, and the induced expression of (A, B) Zhx (A) and Zng (B); (C) separation and purification of Zhx and Zng; mass spectrometry identification of (D, E) Zhx (D) and Zng (E).
FIG. 7 shows the binding site and species specificity analysis of ZHX peptide for hIgG.
Detailed Description
The reagents and materials used in the following examples and experimental examples were commercially available ones unless otherwise specified.
EXAMPLE 1 screening of IgG binding peptides
Because antibodies used in clinic are mainly humanized IgG (human IgG, hIgG) antibodies, in order to obtain a novel hIgG binding domain, in this embodiment, an affinity protein skeleton is used as a basic structure, a library is established by site-directed mutagenesis, and hIgG is used as a bait protein to screen the library, thereby screening hIgG binding peptides (i.e., the IgG binding peptides of the present invention).
1. Construction of a framework protein phage mutation library
In order to obtain the human IgG binding peptide, the invention uses an Affibody (Affibody) as a framework protein (the amino acid sequence of which is shown as SEQ ID NO. 3), and randomly mutates 13 amino acids (Q9, Q10, N11, F13, Y14, L17, H18, E24, E25, R27, N28, Q32 and K35) of the framework protein by using a single-stranded oligonucleotide comprising a plurality of random codons according to FIG. 1. The novel gene resulting from the random mutation was further cloned into a phage vector (containing the carbenicillin resistance gene) according to the method proposed by Kunkel et al (Proc Natl Acad Sci U S A.1985; 82:488-492). Phage mutation libraries were obtained by electroporation (field strength 2.5kV, resistance 200. OMEGA., capacitance 25. Mu.F) into SS3320 helper phage (harboring kanamycin resistance gene) pre-infected E.coli according to the method proposed by Liu et al (NBiotechnol, 2020; 56:46-53).
2. Screening of hIgG binding peptide phage display library
To panning out phage displaying hIgG binding peptides from a phage library of scaffold proteins, hIgG was selected as a bait protein, which was immobilized onto magnetic beads for phage library screening.
The method for fixing hIgG on the magnetic beads is as follows: hIgG was first buffered with phosphate buffer PBS (137mM NaCl,10mM Na) 2 HPO 4 ，2.68mM KCl，1.5mM KH 2 PO 4 ) Diluted to 1mg/ml, biotin was added in a molar ratio of 2 times, and after 1 hour of reaction at room temperature, 20mM arginine and glutamic acid mixture was added to terminate the reaction. The free biotin was removed with a centrifugal desalting column.
Mu.l of biotin-labeled hIgG was mixed with 100. Mu.l of phage, incubated at 4℃overnight or at room temperature for 2 hours, then an appropriate amount of Streptavidin-equipped magnetic beads were added, and incubated at room temperature for 20 minutes in a vertical mixer to capture phage bound to biotin-labeled hIgG. To remove non-specifically bound phages as much as possible, the beads were washed multiple times with washing solution (PBS, 3% bovine serum albumin, 0.05% Tween 20), and finally phages were collected with 500. Mu.l of eluent (50 mM Glycine-HCl, pH 2.7) and transferred to a centrifuge tube with 100. Mu.l Tris-HCl (1M, pH 7.5). The library was amplified by taking a certain amount of phage-infected E.coli. To obtain highly specific hIgG binding peptides, a total of 5 rounds of cycling screening were performed. From the first to fifth rounds of screening, the amount of hIgG was progressively reduced (150, 100, 50, 25 and 12.5 nM) and the number of washes was progressively increased (2, 4,8, 10, 10). After each round of screening, the number of phages was determined by E.coli infection. As shown in FIG. 2A, after phage infection, E.coli was gradient diluted, 10. Mu.l was dropped onto a solid LB plate containing Carbenicillin (100. Mu.g/ml), the plate was tilted to allow the bacterial liquid to flow over the surface of the medium, and then the plate was placed upside down at 37℃for overnight culture, and the colony count was calculated as the amount of positive phage obtained after each round of screening.
The results obtained by the above method are shown in FIG. 2B, and the number of hIgG-binding phage begins to increase after two rounds of screening, and the number of clones per unit reaches 2×10 at the time of the third round of screening 9 About/ml, indicating that phage displaying hIgG binding peptides are enriched by screening. However, during rounds 4 and 5, with further reduction in bait protein, the number of hIgG-binding phage obtained gradually decreased, suggesting that some of the lower affinity phage were eliminated.
To identify whether phage obtained from round 5 of screening displayed hIgG binding peptides, we picked phage monoclonal and analyzed the hIgG binding properties of phage display peptides by ELISA. As a result, as shown in FIG. 3A, we first picked 16 clones (designated C5-1,2,3, …, 16), added to the (hIgG) or non (Iso) hIgG coated ELISA plates, and developed with anti-phage specific antibodies after binding. A blank phage (blk) not displaying the hIgG binding peptide was also used as a negative control for comparison. As a result, it was found that other clones, except for clone No.2 and clone No. 11, were able to bind hIgG, suggesting that these clones displayed hIgG binding peptides. To obtain more hIgG binding peptide information we further extended the screening range and 16 clones (C5-17, 18,19, …, 32) were picked for testing. The results are shown in FIG. 3B, where all 16 clones newly picked were able to bind hIgG, suggesting that these phages displayed hIgG binding peptides.
To obtain the sequence of these phage-displayed hIgG binding peptides, we tested the nucleic acid sequences of 30 clones with significant binding to hIgG and one clone without binding (C5-2), and deduced therefrom the amino acid sequences of these phage-displayed peptides. As a result, as shown in FIG. 4, the amino acid sequences of peptides displayed by 30 phage clones with significant binding to hIgG were identical. Whereas C5-2, which has no hIgG binding properties, exhibits a peptide sequence that differs from the rest of the clone by 9 amino acids. These variant sites are within the scope of the mutations contemplated by the present invention, indicating that the function of these phage display peptides is generated by the mutations contemplated by the present invention. In subsequent tests, phage displayed hIgG binding peptides were designated Zhx and peptides without hIgG binding capacity were designated Zng.
The amino acid sequence of Zhx is shown in SEQ ID NO. 1:
SEQ ID NO.1：
VDNKFNKEWMHAAREIHNLPNLNVYQVVAFIFSLYDDPSQSANLLAEAKKLNDAQAPK。
the amino acid sequence of the framework protein is shown in SEQ ID NO. 3:
VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLAEAKKLNDAQAPK
zng has the amino acid sequence shown in SEQ ID NO. 4:
SEQ ID NO.4：
VDNKFNKEYSNALREIASLPNLNSFQKDAFIWSLDDDPSQSANLLAEAKKLNDAQAPK。
example 2 specific binding of Zhx display phage to hIgG
To further determine the binding sites of the Zhx display phage obtained in example 1 to hIgG, the binding properties of the Zhx display phage to hIgG and fragments thereof (hig-Fc and hig-Fab) were analyzed by ELISA. Meanwhile, in order to determine the species specificity of Zhx, the present example also analyzed the binding of Zhx display phage to murine IgG (mIgG).
The results are shown in FIG. 5, where the Zhx display phage showed dose-dependent binding to hIgG and hIgG-Fc, but not to hIgG-Fab. In the absence of coated hIgG or its fragments (Iso-Zhx), the Zhx display phage could not be enriched in the ELISA plate, indicating that binding was either hIgG or hIgG-Fc dependent. Negative control Zng displayed phage did not bind to these molecules. These results indicate that phage displayed Zhx binds to hIgG by recognizing the Fc fragment. However, under the same conditions, zhx display phage did not bind to mIgG, indicating that Zhx binds to hIgG with species specificity.
EXAMPLE 3 recombinant expression, isolation, purification and characterization of Zhx
In order to obtain the hIgG binding peptide Zhx, the present example obtained the Zhx encoding gene by gene synthesis according to conventional molecular biology methods, and inserted into pQE30 expression vector by introducing BamHI and SalI cleavage sites at both ends of the gene, and then transferred into E.coli M15 strain.
Wherein, the Zhx coding gene is shown in SEQ ID NO. 2:
SEQ ID NO.2：
5’-GTGGACAACAAGTTCAACAAAGAGTGGATGCATGCTGCTAGGGAAA TCCATAATCTGCCGAACCTGAACGTTTATCAGGTTGTTGCTTTCATCTTT TCTCTGTATGATGATCCTAGTCAATCAGCGAACCTGCTGGCGGAAGCGA AAAAACTGAACGATGCGCAGGCGCCGAAA-3’。
to induce expression of Zhx protein, 0.05mM Isopropyl- β -D-thiogalactoside (IPTG) was added to the medium after the expressing strain entered the logarithmic growth phase and induced overnight at 26 ℃,150 rpm. The collected bacteria were resuspended in binding solution (50 mM phosphate, pH8.0, 300mM NaCl,20mM imidazole, 1mM phenylmethylsulfonyl fluoride) and disrupted by high pressure homogenization. The results are shown in FIG. 6A, where Zhx is expressed in E.coli mainly as soluble protein after induction. The purified Zhx product was isolated by Ni-NTA affinity chromatography (FIG. 6C). The mass spectrum identified that the molecular weight of Zhx was 8026Da (fig. 6D), which was completely consistent with the expected molecular weight, shows that the present example gave a perfectly correct sequence of Zhx. In the same way, pure Zng control proteins with the correct sequences were obtained (FIGS. 6B, C, E).
EXAMPLE 4 binding site and specificity analysis of ZHGs by ZHX
To determine the binding site and binding species characteristics of Zhx to hIgG, this example analyzed the binding characteristics of Zhx and Zng prepared in example 3 with reference to the method described in example 2.
The results are shown in FIG. 7, where recombinantly expressed Zhx showed clear dose-dependent binding to hIgG and hIgG-Fc, which did not bind to hIgG-Fab, indicating that Zhx binds hIgG by recognizing the Fc segment. Under conditions without coating hIgG and hIgG-Fc (Iso-Zhx), zhx showed no binding. Negative control Zng showed no binding under all conditions. These results demonstrate that the binding of Zhx is either hIgG or hIgG-Fc dependent. Further analysis found that Zhx did not bind to mIgG, indicating that it was specific for binding to hIgG.
According to the embodiment, through screening of a skeleton protein mutation library, the small molecular peptide Zhx capable of specifically binding to the human IgG through recognition of the Fc segment is obtained, and can be used for drug loading, purification and detection analysis of the human IgG antibody.

Claims (10)

1. A human IgG-binding peptide, characterized by: the amino acid sequence of the polypeptide is shown as SEQ ID NO. 1.
2. The human IgG-binding peptide of claim 1, wherein: it is obtained by expression of the nucleotide sequence shown as SEQ ID NO. 2.
3. Use of a human IgG-binding peptide according to claim 1 or 2 in the preparation of an antibody-loaded medicament.
4. Use according to claim 3, characterized in that: in the antibody-loaded drug, the active small molecule is loaded on IgG through the humanized IgG-binding peptide.
5. Use according to claim 3, characterized in that: the active small molecules are one or the combination of two or more of chemotherapeutics, contrast agents and cytokines.
6. Use according to claim 3, characterized in that: the human IgG-binding peptide and the small active molecule are coupled or fused for expression.
7. An antibody-loaded drug, characterized in that: which is obtained by loading an active small molecule onto IgG by the human IgG-binding peptide according to claim 1 or 2.
8. The antibody-loaded pharmaceutical of claim 7, wherein: the active small molecules are one or the combination of two or more of chemotherapeutics, contrast agents and cytokines.
9. The antibody-loaded pharmaceutical of claim 7, wherein: the human IgG-binding peptide and the small active molecule are coupled or fused for expression.
10. Use of a human IgG-binding peptide according to claim 1 or 2 for purification or detection of IgG.
CN202310050588.0A 2022-04-12 2023-02-01 Human IgG binding peptide and application thereof Pending CN116903712A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202210379886 2022-04-12
CN202210379886X 2022-04-12

Publications (1)

Publication Number Publication Date
CN116903712A true CN116903712A (en) 2023-10-20

Family

ID=88367412

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310050588.0A Pending CN116903712A (en) 2022-04-12 2023-02-01 Human IgG binding peptide and application thereof

Country Status (1)

Country Link
CN (1) CN116903712A (en)

Similar Documents

Publication Publication Date Title
EP4159757A1 (en) Sars-cov-2 spike protein binding molecule and application thereof
CN111848798B (en) Nanometer antibody capable of combining BCMA and application thereof
CN114456260B (en) Novel coronavirus (SARS-COV-2) spike protein binding molecule and application thereof
CN110840830A (en) Injection preparation of anti-CTLA-4 monoclonal antibody
CN114276452A (en) Nano antibody capable of being combined with BCMA (brain cell activating antigen) and application thereof
CN108017712B (en) Protein skeleton derived from shark antibody and application thereof
AU777005B2 (en) Protein A based binding domains with desirable activities
WO2022262101A1 (en) Anti-cd70 antibody having enhanced adcc effect and application thereof
JP6017317B2 (en) Peptide library
WO2024124637A1 (en) Anti-cll1 single-domain antibody and use thereof
CN116903712A (en) Human IgG binding peptide and application thereof
CN113234152B (en) Programmed death receptor-ligand 1 (PD-L1) specific binding polypeptide and application thereof
CN114805559A (en) Fully human anti-new coronavirus receptor binding domain single-chain antibody No4 and application thereof
WO2022037031A1 (en) Il-5 binding molecule, preparation method therefor, and use thereof
CN106928355B (en) CD105 nano antibody Nb184
CN106928358B (en) CD105 nano antibody Nb168
CN106928368B (en) FAP nano antibody Nb57
CN114605526A (en) Single-domain heavy chain antibody aiming at adenovirus vector and application thereof
CN108822189B (en) Specific polypeptide combined with lymphoma cell line in targeted mode and application thereof
CN113461816A (en) Nano antibody aiming at green fluorescent protein GFP and application thereof
US20190264195A1 (en) mRNA DISPLAY ANTIBODY LIBRARY AND METHODS
CN106928359B (en) CD105 nano antibody Nb59
WO2020159524A1 (en) Mrna display antibody library and methods
CN114763379B (en) Specific antibody of new coronavirus S protein, preparation method and application thereof
CN117264061B (en) Nanometer antibody for recognizing uPAR and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination