+

WO2008152537A2 - Anticorps monoclonaux humanisés dirigés contre le récepteur du facteur de croissance épidermique humain, leur utilisation, et procédé correspondant - Google Patents

Anticorps monoclonaux humanisés dirigés contre le récepteur du facteur de croissance épidermique humain, leur utilisation, et procédé correspondant Download PDF

Info

Publication number
WO2008152537A2
WO2008152537A2 PCT/IB2008/052038 IB2008052038W WO2008152537A2 WO 2008152537 A2 WO2008152537 A2 WO 2008152537A2 IB 2008052038 W IB2008052038 W IB 2008052038W WO 2008152537 A2 WO2008152537 A2 WO 2008152537A2
Authority
WO
WIPO (PCT)
Prior art keywords
human
monoclonal antibodies
egfr
humanized monoclonal
humanized
Prior art date
Application number
PCT/IB2008/052038
Other languages
English (en)
Other versions
WO2008152537A3 (fr
Inventor
Mitali Samaddar
Kemburu Prasanna Kumar
Gosala Jaya Lakshmi
Chigurupati Jayaram
Original Assignee
Zenotech Laboratories Limited
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 Zenotech Laboratories Limited filed Critical Zenotech Laboratories Limited
Publication of WO2008152537A2 publication Critical patent/WO2008152537A2/fr
Publication of WO2008152537A3 publication Critical patent/WO2008152537A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C07K16/461—Igs containing Ig-regions, -domains or -residues form different species
    • C07K16/464—Igs containing CDR-residues from one specie grafted between FR-residues from another
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55—Fab or Fab'
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/567—Framework region [FR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present invention is related to humanized MAbs directed against human EGFR that act as receptor antagonists and disrupts the ligand induced signal transduction pathway.
  • the MAbs directed against human EGFR are created using a series of innovative approaches comprising of computational analysis, structure-based modeling studies, a predefined selection criteria, an efficient recombinant DNA expression system and functional interrogation of the MAbs for their therapeutic properties.
  • MAbs Therapeutic monoclonal antibodies
  • HAMA human anti mouse antibody
  • CDR Complementarity Determining Region
  • the human Epidermal Growth Factor Receptor (EGFR and also called Erbl or Herl) belongs to the family of transmembrane, growth factor tyrosine kinase receptors that are important mediators of cell growth, differentiation and survival (Maria F. Structure and activation of EGF receptor. Endocrine Regulations 2002; 36, 87-93).
  • the EGFR signaling pathway plays a crucial role through development and is found to be deregulated in many human cancers. This deregulation is predominantly attributed to EGFR being over-expressed in cancer cells and that is associated with more aggressive clinical behavior, bad prognosis and resistance to chemotherapy. Thus, interference with signaling through the EGFR pathway represents a highly attractive therapeutic approach with potentially broad clinical applications.
  • Such humanized MAbs needs to be expressed in high levels by recombinant DNA technology in order for them to be developed and commercialized as therapeutic drugs.
  • the present invention fulfills these needs in the art and in addition provides an efficient method of humanization and production of MAbs directed against any therapeutic target using recombinant DNA technology.
  • the present invention discloses humanized monoclonal antibodies (MAbs) directed against the therapeutic target, human epidermal growth factor receptor (EGFR) using a series of rationale and innovative processes.
  • the innovative processes comprises of a novel strategy to design humanized MAbs, a rationale approach based on predefined set of selection criteria to identify a subset of humanized MAbs for experimental evaluation and an efficient cloning and recombinant expression strategy to ensure high-level expression of recombinant humanized monoclonal antibodies in mammalian cells.
  • the invention also provides methods to functionally interrogate the recombinant humanized MAbs for their therapeutic properties.
  • the invention provides novel humanized MAbs directed against
  • EGFR that are substantially non-immunogenic and binds to its cognate antigen with high affinity, prevents binding of endogenous ligands and disrupts the human EGFR signaling pathway.
  • the strength of this invention lies in the series of innovative processes that are used for creating and producing humanized monoclonal antibodies.
  • a new strategy and rationale has been applied to generate humanized antibody sequences for a murine MAb directed against human EGFR.
  • CDRs Complementarity Determining Regions
  • this method uses the same parent murine backbone sequences where in the murine framework amino acid residues are substituted rationally with human amino acid residues using a profile of preferred amino acids at specific positions in the human variable region.
  • This position specific amino acid preference profile is created from analysis of multiple sequence alignment of human variable regions available in the protein database.
  • a similar position specific amino acid preference profile is also created for murine variable region so as to identify any highly conserved residues in the frame work regions.
  • Each of the human preferred substitutions is carefully evaluated against their suitability to the local structural properties in the parent murine MAb structure such as location of phi psi angles of residues, packing interfaces of the secondary structural elements, interaction of residues with other residues of CDRs or with other residues in the antibody. From this analysis a panel of humanized sequences is derived and represents a spectrum of 'hard humanization' (with highest possible profile scores) to 'soft humanization' (with low profile scores). These humanized sequences are further evaluated for their structural compatibility using energy minimization followed by molecular dynamic simulations. The structures obtained at the end of the molecular dynamic simulations were compared with the parent structure in order to assess the extent of structural deviations.
  • the humanized MAbs can be produced readily by recombinant DNA technology.
  • This embodiment also describes recombinant expression and extensive characterization of the selected subset of humanized MAbs directed against the human EGFR.
  • the DNA sequences encoding the selected subset of humanized MAbs are appropriately cloned into mammalian expression vectors.
  • the coding sequences are optimized for high-level expression in mammalian cells like CHO (Chinese Hamster Ovary), BHK (Baby Hamster Kidney) or HEK (Human Embryonic Kidney).
  • a gene amplification strategy and a fermentation process are employed to yield expression levels of about lg/litre of spent media. Extensive interrogation of the recombinant anti- EGFR humanized MAbs shows that they will be particularly useful as therapeutic antagonists for EGFR in treating cancers.
  • Figure 1 describes the humanized sequences (variable regions of the light and heavy chains) selected for experimental evaluation.
  • Figure 2 demonstrates SDS-PAGE analysis of purified MAbs under reducing and non-reducing conditions
  • Figure 3 demonstrates immunological identity of the recombinant humanized MAbs and their quantification.
  • Figure 4a shows dose dependent binding of the MAb HZED 8 to purified EGFR
  • Figure 4b shows dose dependent binding of HZED 8 to membrane bound EGFR
  • erbl present on A431 cells.
  • the MAb do not bind to MDA-MB-453 cells known to be negative for erbl.
  • Figure 4c shows specific binding of HZED8 to A431 cells and not to MDA-MB-453 cells by flow cytometric analysis.
  • Figure 4d shows the ability of the MAb HZED 8 to inhibit binding of the endogenous ligand, human EGF to EGFRl present on A431 cells.
  • Figure 5 shows the dose-dependent inhibition of proliferation of A431 cells and not of MDA-MB453 by the MAb HZED8.
  • novel humanized MAbs directed against human EGFR are provided that are substantially non-immunogenic, binds to its cognate antigen with high affinity, prevents binding of endogenous ligands and disrupts the human EGFR signaling pathway.
  • the novel humanized MAbs are created using a series of rationale and innovative processes comprising of computational analysis, structure-based modeling studies, rationale assessment and selection of humanized sequences using predefined selection criteria, an efficient recombinant DNA expression system and studies to functionally interrogate their therapeutic properties.
  • the invention also provides an economic and scalable process of producing the monoclonal antibodies in large quantities and find use in the treatment of human diseases like cancer.
  • the term 'antibody' is a protein consisting of two pairs of immunoglobulin chains, each pair having one light and one heavy chain. In each pair, the light and heavy chain variable regions together form the antigen binding region and the constant regions are responsible for antibody effector functions.
  • An antibody light or heavy chain variable region consists of four 'framework' regions flanking the three 'CDR' regions. The extent of the framework regions and the CDR regions has been defined by Kabat et. al., 1983, Sequences of proteins of Immunological Interest.
  • Chimeric antibodies are those that have been genetically engineered to have the constant region of the heavy and light chain from another species. For example, the constant regions of a murine antibody can be swapped with that of the human antibody to generate a Chimeric antibody.
  • humanized antibody refers to an antibody comprising of human framework regions flanking the CDRs from a non-human (murine) antibody.
  • the constant regions are from human.
  • humanized antibody except for the CDRs is substantially identical to corresponding human antibody.
  • the term monoclonal antibody refers to an antibody that is directed against a specific epitope and is produced by a single B cell or a single hybridoma cell line, which is formed by the fusion of a lymphocyte cell with a myeloma cell.
  • the canonical structure of the CDRs may not be conserved due to interactions with the amino acid residues in the vicinity of the CDRs and thus, may fail to make effective contacts with the cognate antigen similar to those made by CDRs in the parent murine MAb.
  • the amino acid residues in the murine framework regions may make contact with the antigen and contribute to the affinity. Such functionally important residues are lost when humanization is carried out by CDR grafting.
  • the present invention uses a novel approach using a combination of computational analysis, structure-based modeling studies and rational analysis to generate a panel of novel humanized MAbs for the murine MAb M225 (ATCC No. HB-8508) directed against human EGFR, clone and express the same at high levels in mammalian cells using recombinant DNA technology.
  • the first step in the method comprises of determining the structure of the murine MAb using comparative modeling approach. Any standard software like Modeller can be used to determine the structural templates and perform the modeling studies. It is also possible that in some cases that the X-ray crystal structure data is available in the public domain and in such cases the crystal structure can be used for the study.
  • the second step in the method is to identify key residues for substitutions in the murine MAb. This is done by first generating a profile of position specific preferred amino acid residues for human variable region. This profile is aligned with the murine sequence to identify the key residues for substitution. Each of the human preferred substitutions is carefully evaluated for their structural properties in order to allow substitutions of the identified residues in the murine sequence. Finally the structural stability of the humanized sequences was tested using energy minimization studies and molecular dynamic simulations.
  • the present embodiment also describes a method to assess and evaluate the humanized MAbs based on a set of predefined criteria. Modeling studies are carried out to ensure that the selected humanized sequences retained contact interactions with its cognate antigen. This assessment eliminates the humanized sequences that directly or indirectly abrogate the binding interactions.
  • the humanized sequences are scanned for any new potential N-linked or O-linked glycosylation sites.
  • the humanized sequences predicted to have any new glycosylation sites are eliminated as such sequences may have altered binding properties.
  • humanized sequences with minimum root mean square deviation values with reference to the parent structure and also those showing maximum identity to human consensus sequences are selected. Thus a subset of humanized sequences are selected to be carried forward for experimental evaluation based on specific criteria that are important for retaining substantially high binding affinity and also being substantially non-immunogenic.
  • the present embodiment also describes an efficient method of expressing the humanized MAbs in an appropriate mammalian cell host or any other host capable of expressing MAbs in a biologically active form.
  • the DNA sequences encoding the heavy and light chains of the humanized MAbs fused in-frame with appropriate signal sequences are extensively interrogated and designed to achieve high levels of the recombinant MAbs.
  • a gene amplification strategy and a fermentation process are employed to yield expression levels of about lg/litre of spent media. Extensive interrogation of the recombinant anti-EGFR humanized MAbs shows that they will be particularly useful as therapeutic antagonists for EGFR in treating cancers.
  • the method of producing humanized antibodies as described in this invention can also be used to humanize a variety of non-human antibodies.
  • Such humanized antibodies can be used as therapeutic molecules and also for diagnostic purposes.
  • the key amino acid residues in the murine framework regions were identified and rationally substituted with more human like residues.
  • the profile of the human MAb variable region was calculated from the analysis of multiple sequence alignments of the variable regions of known human MAbs from the protein database. The profile was then aligned to the variable regions of the murine MAb. This analysis identified the amino acid residues in the murine framework region that were to be rationally substituted with human preferred residues.
  • Figure 1 describes the humanized sequences (variable regions of the light and heavy chains) selected for experimental evaluation.
  • the DNA coding sequence encoding the variable region of the heavy and light chains of the humanized sequences were analyzed and adapted to codon usage in mammalian cells besides avoiding regions of very high (> 80%) or very low ( ⁇ 30%) GC content. Also during the optimization process other cis-acting sequence motifs like internal TATA-boxes, chi sites, ribosomal entry sites, repeat sequences, RNA secondary structures, ARE (autosomal replicating sequences), INS (inhibitory sequences) or CRS (cis-acting repressive sequence) elements, cryptic splice donor and acceptor sites or branch points are avoided.
  • Such gene optimized sequences are cloned in-frame with their respective human constant regions also optimized using the same procedure as described for the variable regions. More specifically the heavy chain variable region is cloned in-frame with the gamma 1 constant region and the light chain variable region is fused in-frame with the kappa constant region.
  • the human gamma 1 isotype is chosen as it has been found to be the preferred human isotype for supporting antibody dependent cellular toxicity (ADCC) and complement dependent cytotoxicity (CDC) (Riechmann, L et al., (1988) Nature 332:323-327).
  • a signal sequence encoding a signal peptide is fused to the N-terminus of the heavy and light chain coding sequences. The signal peptides have specific cleavage sites that are processed during secretion of the humanized MAbs from the host cell.
  • the assembled coding sequences for the heavy and light chains of the humanized sequences are cloned into a mammalian expression vector containing antibiotic selection markers for bacterial host and the mammalian host.
  • antibiotic selection markers for bacterial host and the mammalian host.
  • the same selection marker can be used for both prokaryotic and the eukaryotic host.
  • the expression vector also contains a selection marker for gene amplification once integrated into the host genome.
  • Hamster Kidney (BHK) cells and Human Embryonic Kidney (HEK) cells are transfected with the constructs containing the humanized sequences for heavy and light chains.
  • a sequential stepwise protocol is used to amplify the loci containing the expression cassettes for high level expression of the humanized MAbs.
  • For each of the humanized sequences stable clones, adapted to serum free media and expressing high levels of the MAbs were carried forward for further investigations.
  • the antibodies were purified from the spent media by protein A affinity chromatography. The eluted antibody was buffer-exchanged into phosphate buffered saline, concentrated, sterile filtered and stored at 4 ° C.
  • the protein A affinity purified MAbs gave a single band of about 150 kDa, as expected, when analyzed by SDS Polyacrylamide Gel Electrophoresis (PAGE) under non-reducing conditions and stained with Coomassie blue dye.
  • the MAbs when analyzed by SDS-PAGE under reducing conditions and stained using Coomassie blue dye showed a band of about 50 kDa as expected for the heavy chain and a second band on about 25 kDa as expected for the light chain.
  • This example provides methods to interrogate the functional properties of the recombinant humanized MAbs directed against the extra cellular domain of human EGFR.
  • the mammalian cell derived humanized MAbs were shown to specifically bind to anti human IgG Fab fragment specific antibody and also to anti-human IgG Fc gamma fragment that is used for detecting the bound MAbs in a sandwich ELISA.
  • Human IgG is used as the reference standard for quantification of the MAbs ( Figure 3).
  • the recombinant humanized MAbs could specifically bind to purified EGFR (erbl) from human carcinoma A431 cells shown using ELISA and also directly to membrane bound EGFR (erbl) present on A431 cells shown using cell based assay and flow cytometry.
  • the human breast cancer cell line MDA-MB-453 known to be negative for erbl is used as a negative control in the experiment.
  • the recombinant humanized MAbs does not bind to MDA-MB-453 cells as shown by cell based assay and also flow cytometry.
  • the binding of the humanized MAbs to the EGFRl receptors present on A431 cells also inhibits binding of the endogenous ligand human EGF.
  • the EC50 values (101 ng/ml for HZED8 and 91 ng/ml for 225) for the receptor binding experiment (Fig. 4b) and IC50 values (0.26 nM for HZED8 and 0.29 nM for 225) for the ligand inhibition assay (Fig. 4d) are comparable for HZED8 and the reference anti-EGFR MAb 225. More over the data also shows that the MAb HZED8 is highly specific and does not have crossreactivity with a cell line that is negative for the EGFRl receptor.
  • Figure 4a shows dose dependent binding of HZED8 to purified EGFRl when immobilized onto high-binding ELISA plates.
  • Figure 4b shows dose dependent binding of MAbs to membrane bound EGFR (erbl) present on A431 cells.
  • the MAbs do not bind to MDA-MB-453 cells known to be negative for erbl.
  • Figure 4c shows specific binding of the MAbs to A431 cells and not to MDA-
  • Figure 4d shows the ability of HZED8 to inhibit binding of the endogenous ligand human EGF to EGFRl present on A431 cells.
  • FIG. 5 shows the dose-dependent inhibition of proliferation of A431 cells and not of MDA-MB453 by recombinant humanized MAbs.
  • the EC50 value for HZED8 is 128 ng/ml and is comparable to that for the reference MAb 225 (116 ng/ml).
  • the experiment also shows specificity of HZED8 in terms of its biological activity as it fails to inhibit proliferation of the MDA-MB-453 cells.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

La présente invention concerne des anticorps monoclonaux humanisés conçus, créés et produits au moyen d'une série de processus novateurs impliquant l'analyse computationnelle, études de modélisation, vérification et sélection rationnelle de séquences humanisées, un système efficace d'expression d'ADN recombinant et des études fonctionnelles visant à connaître en profondeur les propriétés des anticorps monoclonaux. L'invention propose également un processus économique et d'envergure variable permettant de produire de grandes quantités d'anticorps monoclonaux et de s'appliquer au traitement de maladies humaines telles que le cancer. L'invention concerne enfin ensemble de référence d'anticorps monoclonaux humanisés de l'invention dirigés contre le récepteur du facteur de croissance épidermique humain, agissant comme antagonistes des récepteur, et sensiblement non immunogènes.
PCT/IB2008/052038 2007-06-14 2008-05-23 Anticorps monoclonaux humanisés dirigés contre le récepteur du facteur de croissance épidermique humain, leur utilisation, et procédé correspondant WO2008152537A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN1236CH2007 2007-06-14
IN1236/CHE/2007 2007-06-14

Publications (2)

Publication Number Publication Date
WO2008152537A2 true WO2008152537A2 (fr) 2008-12-18
WO2008152537A3 WO2008152537A3 (fr) 2010-04-08

Family

ID=40130263

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2008/052038 WO2008152537A2 (fr) 2007-06-14 2008-05-23 Anticorps monoclonaux humanisés dirigés contre le récepteur du facteur de croissance épidermique humain, leur utilisation, et procédé correspondant

Country Status (1)

Country Link
WO (1) WO2008152537A2 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013134743A1 (fr) 2012-03-08 2013-09-12 Halozyme, Inc. Anticorps anti-récepteur du facteur de croissance épidermique conditionnellement actifs et leurs procédés d'utilisation
WO2015038984A2 (fr) 2013-09-12 2015-03-19 Halozyme, Inc. Anticorps anti-récepteur du facteur de croissance épidermique modifiés et procédés pour les utiliser
US9072798B2 (en) 2009-02-18 2015-07-07 Ludwig Institute For Cancer Research Ltd. Specific binding proteins and uses thereof
WO2017161206A1 (fr) 2016-03-16 2017-09-21 Halozyme, Inc. Conjugués contenant des anticorps à activité conditionnelle ou des fragments de liaison à un antigène associés, et procédés d'utilisation

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6114146A (en) * 1994-11-14 2000-09-05 Baxter Aktiengesellschaft Expression plasmid, a fusion protein, a transfected eukaryotic cell line, a method of producing foreign proteins, a foreign protein preparation as well as a pharmaceutical composition
US6602684B1 (en) * 1998-04-20 2003-08-05 Glycart Biotechnology Ag Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6114146A (en) * 1994-11-14 2000-09-05 Baxter Aktiengesellschaft Expression plasmid, a fusion protein, a transfected eukaryotic cell line, a method of producing foreign proteins, a foreign protein preparation as well as a pharmaceutical composition
US6602684B1 (en) * 1998-04-20 2003-08-05 Glycart Biotechnology Ag Glycosylation engineering of antibodies for improving antibody-dependent cellular cytotoxicity

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ROGUSKA ET AL.: 'Humanization of murine monoclonal antibodies through variable domain resurfacing' PNAS vol. 91, 1994, pages 969 - 973 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9072798B2 (en) 2009-02-18 2015-07-07 Ludwig Institute For Cancer Research Ltd. Specific binding proteins and uses thereof
WO2013134743A1 (fr) 2012-03-08 2013-09-12 Halozyme, Inc. Anticorps anti-récepteur du facteur de croissance épidermique conditionnellement actifs et leurs procédés d'utilisation
EP3296320A1 (fr) 2012-03-08 2018-03-21 Halozyme, Inc. Facteur de croissance anti-épidermique actif de manière conditionnelle des anticorps de récepteurs et leurs procédés d'utilisation
WO2015038984A2 (fr) 2013-09-12 2015-03-19 Halozyme, Inc. Anticorps anti-récepteur du facteur de croissance épidermique modifiés et procédés pour les utiliser
WO2017161206A1 (fr) 2016-03-16 2017-09-21 Halozyme, Inc. Conjugués contenant des anticorps à activité conditionnelle ou des fragments de liaison à un antigène associés, et procédés d'utilisation

Also Published As

Publication number Publication date
WO2008152537A3 (fr) 2010-04-08

Similar Documents

Publication Publication Date Title
JP7538721B2 (ja) 抗インターロイキン１７ａ抗体、医薬組成物、およびその使用
CN113416258B (zh) 一种多特异性抗体及其制备方法和用途
CN109922864B (zh) 有共同轻链的双互补位和多互补位抗体和使用方法
CN114746440B (zh) 新型多肽复合物
CN115960232A (zh) 抗ctla4的单克隆抗体或其抗原结合片段、药物组合物及用途
WO2018158719A1 (fr) Protéines hétérodimères modifiées
KR20080039844A (ko) Ｂ―세포 림프종 치료를 위한 ｃｄ20에 대한 모노클로날항체의 제조 방법
CN106397598B (zh) 多价多特异性抗体及免疫杂合蛋白的表达和制备方法
WO2007108152A1 (fr) Anticorps bispecifique hautement fonctionnel
EP3247721A1 (fr) Anticorps igg bispécifiques et leurs procédés de préparation
WO2004085474A2 (fr) Procede de production d'un anticorps contre le recepteur du facteur de croissance epidermique
WO2015032916A1 (fr) Anticorps dirigés contre des cibles complexes
WO2015146437A1 (fr) ANTICORPS BISPÉCIFIQUE DE L'IgG2 HAUTEMENT FONCTIONNEL
CA3211407A1 (fr) Anticorps trispecifique anti-gprc5d x bcma x cd3 et son utilisation
US20090220520A1 (en) Recombinant method for the production of a monoclonal antibody to CD52 for the treatment of chronic lymphocytic leukemia
EP3728328B1 (fr) Construction bispécifique de liaison à un antigène
EP4303231A1 (fr) Anticorps bispécifique
EP3116906A1 (fr) Anticorps anti-laminine 4 spécifiques de lg1-3
WO2008152537A2 (fr) Anticorps monoclonaux humanisés dirigés contre le récepteur du facteur de croissance épidermique humain, leur utilisation, et procédé correspondant
EP1622941A2 (fr) Procede de production d'un anticorps contre le recepteur du facteur de croissance epidermique
US20220306741A1 (en) Purification Method for Bispecific antigen-binding Polypeptides with Enhanced Protein L Capture Dynamic Binding Capacity
JP6210455B2 (ja) 剛直なヘリックス構造を有するオリゴペプチドが導入された二重特異性抗体
CN119137147A (zh) 具有增加的选择性的多链多靶向性双特异性抗原结合分子
CN114269788A (zh) 一种能够与人4-1bb结合的分子及其应用
CN115073596A (zh) 一种人源化Claudin 18.2抗体及其应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08751313

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08751313

Country of ref document: EP

Kind code of ref document: A2

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载