共查询到20条相似文献,搜索用时 296 毫秒
1.
Renner C Stehle I Lee FT Hall C Catimel B Nice EC Mountain A Rigopoulos A Brechbiel MW Pfreundschuh M Scott AM 《Cancer immunology, immunotherapy : CII》2001,50(2):102-108
Bispecific antibodies are currently being used in clinical trials in increasing numbers in the areas of breast cancer, prostate
cancer, non-Hodgkin's lymphoma and Hodgkin's lymphoma. We have previously performed two clinical trials in patients with Hodgkin's
disease with an anti-CD30/anti-CD16 bispecific antibody and demonstrated a 30% response rate in a cohort of patients otherwise
resistant to standard therapeutic modalities. However, no surrogate marker could be defined in these trials indicative of
optimal antibody dosing/scheduling or predictive for favorable response. In order to evaluate accurately the potential biodistribution
properties of bispecific antibody in patients, we have performed a detailed analysis of the binding properties and animal
model in vivo characteristics of these constructs. For this purpose, the parental antibodies (anti-CD30 and anti-CD16) and
the bispecific antibody (anti-CD30/anti-CD16) were radiolabeled with either 125I or 111In. Antibody integrity and binding properties after labeling were confirmed by Scatchard plot and Lindmo analysis. 111In-labeled antibodies revealed superior targeting properties in a standard SCID mouse tumor model. Both the bivalent parental
anti-CD30 monoclonal antibody and the monovalent anti-CD30/anti-CD16 bispecific antibody showed excellent uptake in CD30+ tumors which did not differ significantly between the two (maximum uptake 16.5% ± 4.2% vs. 18.4% ± 3.8% injected dose/gram
tissue). The equivalent targeting properties of the bispecific antibody compared with the parental anti-CD30 antibody encourages
the further clinical development of this bispecific antibody, and might help to explain the clinical responses seen with this
antibody so far in patients suffering from Hodgkin's disease.
Received: 26 October 2000 / Accepted: 15 December 2000 相似文献
2.
Background
A natural bispecific antibody, which can be produced by exchanging Fab arms of two IgG4 molecules, was first described in allergic patients receiving therapeutic injections with two distinct allergens. However, no information has been published on the production of natural bispecific antibody in animals. Even more important, establishment of an animal model is a useful approach to investigate and characterize the naturally occurring antibody.Methodology/Principal Findings
We demonstrated that a natural bispecific antibody can also be generated in New Zealand white rabbits by immunization with synthesized conjugates. These antibodies showed bispecificity to the components that were simultaneously used to immunize the animals. We observed a trend in our test animals that female rabbits exhibited stronger bispecific antibody responses than males. The bispecific antibody was monomeric and primarily belonged to immunoglobulin (Ig) G. Moreover, bispecific antibodies were demonstrated by mixing 2 purified monospecific antibodies in vivo and in vitro.Conclusions/Significance
Our results extend the context of natural bispecific antibodies on the basis of bispecific IgG4, and may provide insights into the exploration of native bispecific antibodies in immunological diseases. 相似文献3.
de Boisferon Marc Hillairet Manetti Corrine Raguin Olivier Gautherot Emmanuel Rostène William Barbet Jacques Gruaz-Guyon Anne 《International journal of peptide research and therapeutics》1997,4(4-6):331-339
Summary The advantages of bivalent hapten-bearing peptides for the detection of tumours pretargeted with bispecific antibodies have
been demonstrated. This technology is now considered for radioimmunotherapy and bivalent haptens designed to target131I are needed. We thus synthesised a series of tyrosine-containing peptides bearing the histamine-hemisuccinate hapten. These
molecules were tested for their ability to bind simultaneously two anti-hapten antibody molecules. One of these bivalent haptens,
AG3.0, with a lysyl-d-tyrosyl-lysine connecting chain, was found to have optimal binding characteristics and was thus selected for further investigations.
AG3.0 was shown to efficiently deliver radioactive iodine to human colorectal tumours grafted in nude mice using an anti-carcinoembryonic
antigen×anti-histamine-hemisuccinate bispecific antibody. AG3.0 was also targeted to human B lymphoma cells pretargeted with
a bispecific antibody specific for membrane IgM. In this system, bivalent ligands such as F(ab′)2 or IgG are rapidly internalised and covalently linked radioactive iodine is released from target cells as a result of intracellular
catabolism. With the pretargeted iodine-labelled bivalent hapten, a fivefold increase in the intracellular activity retention
time as compared to125I-labelled F(ab′)2 and IgG was observed. The radiolabelled hapten did not undergo any degradation after internalisation. These results have
been confirmed in vivo with an anti-BCL1 IgM idiotype bispecific antibody and131I-labelled AG3.0. These reagents injected as a single 300 μCi dose, 7 days after inoculation of 104 BCL1 lymphoma cells in BALB/c mice, cured 14/16 of the animals and the treatment was well tolerated. Comparatively, the same dose
of labelled IgG cured 13/16 of the mice but three mice died of haematologic toxicity. The same dose of labelled F(ab′)2 or Fab′ was completely inefficient.131I-labelled bivalent haptens are now used in phase I radioimmunotherapy clinical trials. 相似文献
4.
James E. Wooldridge Chris E. Dahle George J. Weiner 《Cancer immunology, immunotherapy : CII》1997,45(3-4):174-179
T cells play a key role in the control of abnormal B cell proliferation. Factors that play a role in inadequate T cell responses
include absence of expression of costimulatory and adhesion molecules by the malignant B cells and lack of cytotoxic T cells
specific for tumor-associated antigens. A number of approaches have been used to enhance T cell response against malignant
B cells. Agents such as soluble CD40 ligand can enhance expression of costimulatory molecules by the malignant B cells and
improve their ability to activate T cells. Anti-CD3-based bispecific antibodies can retarget T cells toward the tumor cells
irrespective of T cell specificity. We used the V 38C13 murine lymphoma model to assess whether the combination of soluble
CD40 ligand and anti-CD3-based bispecific antibody can enhance T cell activation induced by malignant B cells more effectively
than either approach alone. Expression of CD80, CD86, and ICAM-1 on lymphoma cells was up-regulated by soluble CD40 ligand.
Syngeneic T cells were activated more extensively by lymphoma cells when the lymphoma cells were pre-treated with soluble
CD40 ligand. Bispecific-antibody induced T cell activation was more extensive when lymphoma cells pretreated with soluble
CD40 ligand were present. The combination of soluble CD40 ligand plus bispecific antibody enhanced the median survival of
mice compared to mice treated with bispecific anibody alone. We conclude that pretreatment of tumor cells with agents capable
of inducing costimulatory molecule expression, such as soluble CD40 ligand can enhance the ability of malignant B cells to
activate T cells. This effect is enhanced by the addition of bispecific antibody. The combination of enhanced expression of
costimulatory molecules and retargeting of T cells by bispecific antibody may allow for a more effective T-cell-based immunotherapy.
Accepted: 14 October 1997 相似文献
5.
Pullarkat V Deo Y Link J Spears L Marty V Curnow R Groshen S Gee C Weber JS 《Cancer immunology, immunotherapy : CII》1999,48(1):9-21
A phase I study of escalating doses of humanized bispecific antibody (bsAb) MDX-H210 with granulocyte-colony-stimulating
factor (G-CSF) was conducted in patients with metastatic breast cancer that overexpressed HER2/neu. The main objectives of
the study were to define the maximal tolerated dose (MTD) of MDX-H210 when combined with G-CSF, to measure the pharmacokinetics
of MDX-H210 when administered with G-CSF, and to determine the toxicity, biological effects and possible therapeutic effect
of MDX-H210 with G-CSF. MDX-H210 is a F(ab)′ × F(ab)′ humanized bispecific murine antibody that binds to both HER2/neu and
the FcγR1 receptor (CD64), and was administered intravenously weekly for three doses followed by a 2-week break and then three
more weekly doses. A total of 23 patients were treated, and doses were escalated from 1 mg/m2 to 40 mg/m2 with no MTD reached. The toxicity of the bsAb + G-CSF combination was modest, with no dose-limiting toxicity noted: 19 patients
had fevers, 7 patients had diarrhea, and 3 patients had allergic reactions that did not limit therapy. The β-elimination half-life
varied from 4 h to 8 h at doses up to 20 mg/m2. Significant release of cytokines interleukin-6, G-CSF, and tumor necrosis factor α was observed after administration of
bsAb. Circulating monocytes disappeared within 1 h of bsAb infusion, which correlated with binding of bsAb, noted by flow-cytometric
analysis. Significant levels of human anti-(bispecific antibody) were measured in the plasma of most patients by the third
infusion. No objective clinical responses were seen in this group of heavily pre-treated patients.
Received: 23 July 1998 / Accepted: 6 November 1998 相似文献
6.
靶向人类免疫缺陷病毒(human immunodeficiency virus,HIV)流行毒株的广谱中和抗体(broadly neutralizing antibody, bNAb)单一疗法最终会导致机体出现病毒逃逸突变,然而基于广谱中和抗体开发的双特异性或多特异性抗体则表现出较好的中和效力和广谱性。根据已公布的单链可变区基因抗体序列,经密码子优化后合成一种由单基因编码的双特异性抗体iMab-PGT151,经双酶切和测序对重组质粒进行了验证。酶联免疫吸附试验检测双特异性抗体的结合特异性,检测U87细胞裂解液中的荧光素酶活性以定量分析双特异性抗体对HIV-1假病毒的中和作用;间接免疫荧光染色法检测双特异性抗体iMab-PGT151对人喉癌上皮细胞的反应性;酶联免疫吸附试验检测该抗体对心磷脂的结合能力,验证其自体反应性。结果显示,构建的双特异抗体iMab-PGT151能够成功表达,可分别结合亲本抗体的各个配体,具有双特异性,能100%中和20株假病毒,IC50值为0.084 μg/mL。与亲本抗体相比,该抗体具有更强的中和效力和广谱度,无自体反应性,具有临床适用性。所构建的双特异性抗体iMab-PGT151将可能成为预防和治疗HIV-1感染的有效候选药物之一。 相似文献
7.
Bernd J. Wranik Erin L. Christensen Gabriele Schaefer Janet K. Jackman Andrew C. Vendel Dan Eaton 《The Journal of biological chemistry》2012,287(52):43331-43339
The ability of bispecific antibodies to simultaneously bind two unique antigens has great clinical potential. However, most approaches utilized to generate bispecific antibodies yield antibody-like structures that diverge significantly from the structure of archetype human IgG, and those that do approach structural similarity to native antibodies are often challenging to engineer and manufacture. Here, we present a novel platform for the mammalian cell production of bispecific antibodies that differ from their parental mAbs by only a single point mutation per heavy chain. Central to this platform is the addition of a leucine zipper to the C terminus of the CH3 domain of the antibody that is sufficient to drive the heterodimeric assembly of antibody heavy chains and can be readily removed post-purification. Using this approach, we developed various antibody constructs including one-armed Abs, bispecific antibodies that utilize a common light chain, and bispecific antibodies that pair light chains to their cognate heavy chains via peptide tethers. We have applied this technology to various antibody pairings and will demonstrate the engineering, purification, and biological activity of these antibodies herein. 相似文献
8.
With more than 30 therapeutic monoclonal antibodies (mAbs) approved and annual global sales of the products at ∼$50 billion in 2010, these products have proven to be successful in many ways. Nevertheless, there is room for improvement in performance, and substantial unmet medical needs remain. As a consequence, numerous organizations are devoting resources to engineering novel mAbs such as bispecific antibodies that have increased functionality compared with unmodified IgG molecules. The World Bispecific Antibody Summit, organized by Hanson Wade, drew over 100 participants to Boston to discuss engineering novel bispecific antibodies, generating lead candidates and clinical study and commercialization of the molecules. Approaches such as the trifunctional antibody (TRION), dual variable domain-Ig (Abbott), two-in-one (Genentech), dual affinity retargeting (MacroGenics), kappa-lambda body (NovImmune), bispecific T-cell engager (Micromet) and chemical generation (CovX/Pfizer) were discussed in detail. In addition, posters describing bispecific Affibody® molecules for targeting of EGFR and HER2 (Affibody), T-cell receptor based bi-specifics that target HLA-peptides (Immunocore), a novel mAb-Fv bispecific antibody format utilizing Fc region (Xencore), generation of a tetravalent bispecific antibody against IL4 and IL13 for the treatment of idiopathic pulmonary fibrosis (Sanofi), Combining Affibody® molecules and the AlbumodTM technology to create long acting multispecific protein therapeutics (Royal Institute of Technology, Affibody) and COVA301 as a highly potent bispecific inhibitor of IL-17A and TNFα (Covagen) were presented.Key words: bispecific antibodies, antibody engineering, therapeutic antibodies 相似文献
9.
T-cell activation and B-cell depletion in chimpanzees treated with a bispecific anti-CD19/anti-CD3 single-chain antibody construct 总被引:4,自引:4,他引:0
Schlereth B Quadt C Dreier T Kufer P Lorenczewski G Prang N Brandl C Lippold S Cobb K Brasky K Leo E Bargou R Murthy K Baeuerle PA 《Cancer immunology, immunotherapy : CII》2006,55(5):503-514
BscCD19xCD3 is a bispecific single-chain antibody construct with exceptional cytotoxic potency in vitro and in vivo. Here, we have investigated the biological activity of bscCD19xCD3 in chimpanzee, the only animal species identified in which bscCD19xCD3 showed bispecific binding, redirected B-cell lysis and cytokine production comparable to human cells. Pharmacokinetic analysis following 2-h intravenous infusion of 0.06, 0.1 or 0.12 μg/kg of bscCD19xCD3 as part of a dose escalation study in a single female chimpanzee revealed a half-life of approximately 2 h and elimination of the bispecific antibody from circulation within approximately 8 h after the end of infusion. This short exposure to bscCD19xCD3 elicited a transient increase in serum levels of IFNγ, IL-6, IL-2, soluble CD25, and transiently upregulated expression of CD69 and MHC class II on CD8-positive cells. Cytokine release and upregulation of T-cell activation markers were not observed with vehicle controls. A multiple-dose study using 5 weekly doses of 0.1 μg/kg in two animals also showed transient cytokine release and an activation of peripheral T cells with a first-dose effect, accompanied by a transient lymphopenia. While oscillations of T-cell counts were relatively even during repeated treatments, the amplitudes of peripheral B cells declined with every infusion, which was not observed in a vehicle control animal. Our data show that bscCD19xCD3 can be safely administered to chimpanzees at dose levels that cause fully reversible T-cell activation and, despite a very short exposure time, cumulative loss of peripheral B lymphocytes. A clinical trial testing prolonged administration of bscCD19xCD3 (MT103) for improving efficacy is currently ongoing. 相似文献
10.
Antibody-directed fibrinolysis. An antibody specific for both fibrin and tissue plasminogen activator 总被引:3,自引:0,他引:3
C Bode M S Runge E E Branscomb J B Newell G R Matsueda E Haber 《The Journal of biological chemistry》1989,264(2):944-948
An investigation was made to determine whether it is possible to attract tissue plasminogen activator (tPA) to the site of a thrombus by means of an antibody with affinites for both tPA and fibrin. A bispecific antibody conjugate was constructed by cross-linking two monoclonal antibodies: one specific for tPA, the other specific for fibrin. The bispecific antibody enhanced fibrinolysis by capturing tPA at the site of a fibrin deposit. In an in vitro quantitative fibrinolysis assay, the relative fibrinolytic potency of tPA bound to the bispecific antibody was 13 times greater than that of tPA and 200 times greater than that of urokinase. When fibrin was treated with the bispecific antibody before being mixed with tPA, the relative fibrinolytic potency of tPA was enhanced 14-fold. This capture also occurred when the concentration of tPA present in the assay was less than the concentration of tPA present in normal human plasma. In a human plasma clot assay, samples containing both the bispecific antibody and tPA exhibited significantly more lysis than did samples containing tPA alone. In spite of the increased clot lysis effected by the presence of bispecific antibody, there was no significant increase in fibrinogen or alpha 2-antiplasmin degradation at equal tPA concentrations. The ability of the bispecific antibody to concentrate exogenous tPA in vivo was then examined in the rabbit jugular vein model. Systemic infusion of a small amount of tPA (10,000 units) produced no significant increment in thrombolysis over the level of spontaneous lysis (14 +/- 8%). However, the simultaneous infusion of 10,000 units of tPA and 2 mg of bispecific antibody resulted in 42 +/- 14% (p less than 0.01) lysis. These results suggest that a molecule capable of binding both fibrin and tPA with high affinity could enhance thrombolysis in the circulation by capturing endogenous tPA. 相似文献
11.
《MABS-AUSTIN》2013,5(8):1248-1259
ABSTRACTBispecific antibody therapeutics can expand the functionality of a conventional monoclonal antibody drug because they can bind multiple antigens. However, their great potential is counterbalanced by the challenges faced in their production. The classic asymmetric bispecific containing an Fc requires the expression of four unique chains – two light chains and two heavy chains; each light chain must pair with its correct heavy chain, which then must heterodimerize to form the full bispecific. The light-chain pairing problem has several solutions, some of which require engineering and optimization for each bispecific pair. Here, we introduce a technology called EFab Domain Substitution, which replaces the Cε2 of IgE for one of the CL/CH1 domains into one arm of an asymmetric bispecific to encourage the correct pairing of the light chains. EFab Domain Substitution provides very robust correct pairing while maintaining antibody function and is effective for many variable domains. We report its effect on the biophysical properties of an antibody and the crystal structure of the EFab domain substituted into the adalimumab Fab (PDB ID 6CR1). 相似文献
12.
Schlereth B Kleindienst P Fichtner I Lorenczewski G Brischwein K Lippold S da Silva A Locher M Kischel R Lutterbüse R Kufer P Baeuerle PA 《Cancer immunology, immunotherapy : CII》2006,55(7):785-796
Bispecific single-chain antibody constructs specific for human CD3 have been extensively studied for antitumor activity in
human xenograft models using severe combined immunodeficient mice supplemented with human T cells. High efficacy at low effector-to-target
ratios, independence of T cell costimuli and a potent activation of previously unstimulated polyclonal T cells were identified
as hallmarks of this class of bispecific antibodies. Here we studied a bispecific single-chain antibody construct (referred
to as ‘bispecific T cell engager’, BiTE) in an immunocompetent mouse model. This was possible by the use of a murine CD3-specific
BiTE, and a syngeneic melanoma cell line (B16F10) expressing the human Ep-CAM target. The murine CD3-specific BiTE, called
2C11x4-7 prevented in a dose-dependent fashion the outgrowth of subcutaneously growing B16/Ep-CAM tumors with daily i.v. injections
of 5 or 50 μg BiTE which was most effective. Treatment with 2C11x4-7 was effective even when it was started 10 days after
tumor cell inoculation but delayed treatments showed a reduction in the number of cured animals. 2C11x4-7 was also highly
active in a lung tumor colony model. When treatment was started on the day of intravenous tumor cell injection, seven out
of eight animals stayed free of lung tumors, and three out of eight animals when treatment was started on day 5. Our study
shows that BiTEs also have a high antitumor activity in immunocompetent mice and that there is no obvious need for costimulation
of T cells by secondary agents.
Bernd Schlereth and Petra Kleindienst contributed equally to this work. 相似文献
13.
Michael J Gramer Ewald TJ van den Bremer Muriel D van Kampen Amitava Kundu Peter Kopfmann Eric Etter David Stinehelfer Justin Long Tom Lannom Esther H Noordergraaf Jolanda Gerritsen Aran F Labrijn Janine Schuurman Patrick HC van Berkel Paul WHI Parren 《MABS-AUSTIN》2013,5(6):962-973
The manufacturing of bispecific antibodies can be challenging for a variety of reasons. For example, protein expression problems, stability issues, or the use of non-standard approaches for manufacturing can result in poor yield or poor facility fit. In this paper, we demonstrate the use of standard antibody platforms for large-scale manufacturing of bispecific IgG1 by controlled Fab-arm exchange. Two parental antibodies that each contain a single matched point mutation in the CH3 region were separately expressed in Chinese hamster ovary cells and manufactured at 1000 L scale using a platform fed-batch and purification process that was designed for standard antibody production. The bispecific antibody was generated by mixing the two parental molecules under controlled reducing conditions, resulting in efficient Fab-arm exchange of >95% at kg scale. The reductant was removed via diafiltration, resulting in spontaneous reoxidation of interchain disulfide bonds. Aside from the bispecific nature of the molecule, extensive characterization demonstrated that the IgG1 structural integrity was maintained, including function and stability. These results demonstrate the suitability of this bispecific IgG1 format for commercial-scale manufacturing using standard antibody manufacturing techniques. 相似文献
14.
Franziska Hirschhaeuser Stefan Walenta Wolfgang Mueller-Klieser 《Cancer immunology, immunotherapy : CII》2010,59(11):1675-1684
Catumaxomab is an intact trifunctional bispecific antibody targeting human EpCAM (epithelial cell adhesion molecule) and CD3
with further binding to Fcγ receptor type I, IIa and III. We choose multicellular tumor spheroids (MCTS) of human EpCAM-positive
FaDu tumor cells in co-culture with human peripheral blood mononuclear cells as an adequate three-dimensional in vitro model
for pharmacological testing of catumaxomab. We found a strong dose-dependent antitumor response mediated by catumaxomab, with
volume-decreased or completely destroyed tumor spheroids together with a massive immune cell infiltration and decreased signals
for cancer cell viability and clonogenicity. In control experiments with F(ab′)2 fragments of catumaxomab and the parental
antibodies alone or in combination the effects in spheroid volume reduction were less than that of catumaxomab. All binding
partners of the postulated tricell complex have to be present to exert catumaxomab’s full mode of action. These distinct effects
of catumaxomab are based on the unique composition of the trifunctional bispecific antibody. Since, in general, many cancers
are treated by chemotherapy in combination with immunological tumor therapy, we additionally analyzed the effects of cisplatin
alone and in combination with catumaxomab. For cisplatin alone we detected a dose-dependent response relating to decrease
of spheroid volume. The combined approach resulted in a synergistic spheroid volume decrease and the colony formation was
reduced to non-detectable levels. 相似文献
15.
《MABS-AUSTIN》2013,5(1):4-13
With more than 30 therapeutic monoclonal antibodies (mAbs) approved and annual global sales of the products at ~$50 billion in 2010, these products have proven to be successful in many ways. Nevertheless, there is room for improvement in performance, and substantial unmet medical needs remain. As a consequence, numerous organizations are devoting resources to engineering novel mAbs such as bispecific antibodies that have increased functionality compared with unmodified IgG molecules. The World Bispecific Antibody Summit, organized by Hanson Wade, drew over 100 participants to Boston to discuss engineering novel bispecific antibodies, generating lead candidates and clinical study and commercialization of the molecules. Approaches such as the trifunctional antibody (TRION), dual variable domain-Ig (Abbott), two-in-one (Genentech), dual affinity retargeting (MacroGenics), kappa-lambda body (NovImmune), bispecific T-cell engager (Micromet) and chemical generation (CovX/Pfizer) were discussed in detail. In addition, posters describing bispecific Affibody® molecules for targeting of EGFR and HER2 (Affibody), T-cell receptor based bi-specifics that target HLA-peptides (Immunocore), a novel mAb-Fv bispecific antibody format utilizing Fc region (Xencore), generation of a tetravalent bispecific antibody against IL4 and IL13 for the treatment of idiopathic pulmonary fibrosis (Sanofi), Combining Affibody® molecules and the Albumod? technology to create long acting multispecific protein therapeutics (Royal Institute of Technology, Affibody) and COVA301 as a highly potent bispecific inhibitor of IL-17A and TNF-α (Covagen) were presented. 相似文献
16.
17.
Clearance of blood-borne pathogens mediated through bispecific monoclonal antibodies bound to the primate erythrocyte complement receptor 总被引:1,自引:0,他引:1
R. P. Taylor Alessandra Nardin William M. Sutherland 《Cancer immunology, immunotherapy : CII》1997,45(3-4):152-155
The primate erythrocyte complement receptor facilitates both the immune adherence reaction and the immune complex clearance
properties of primate erythrocytes. These phenomena have been studied for more than 40 years. However, it has only recently
become apparent that these characteristics of primate erythrocytes may be useful in the generation of a therapy based on bispecific
monoclonal antibodies. Our approach uses bispecific monoclonal antibody constructs (heteropolymers) that promote binding of
specific target pathogens to primate erythrocytes via the complement receptor. Once bound to the erythrocytes, the pathogen-heteropolymer complex should be cleared from the circulation,
phagocytosed and destroyed in the liver. Results with several prototype target pathogens in monkey models indicate it may
be possible to use this technology to develop a robust and general therapy for the treatment of diseases associated with blood-borne
pathogens.
Accepted: 14 October 1997 相似文献
18.
Y Tsukada K Ohkawa N Hibi K Tsuzuki K Oguma H Satoh 《Cancer biochemistry biophysics》1989,10(3):247-256
A monoclonal mouse antibody (MoHG) was produced using in vitro cultured AH66R tumor cells treated with cholesteryl hemisuccinate as an immunogen. The antibody identified a 90 kd membrane glycoprotein (HG-90) which is expressed on in vitro cultured hepatoma cell lines AH66 and AH66R. A monoclonal antibody was prepared to the anthracycline drug daunomycin, and it also reacted with adriamycin. A fusion was made of the hybridoma HG-90 with the hybridoma which recognized daunomycin/adriamycin. This bispecific hybridoma A8C recognized both determinants. We studied the therapeutic effect of the A8C bispecific antibody with adriamycin treatment and compared it to the effect of the bispecific antibody to which adriamycin had been conjugated via an albumin (Alb) bridge. The therapy model used was the tumor AH66R in Donryu rats. Tumor bearing rats had their subcutaneous tumors resected on day 10, a time when distant metastases were present. After the surgical resection of the tumor the rats were injected intravenously for two cycles with the bispecific antibodies, followed by the administration of adriamycin (ADR) or MoHG.Alb.ADR conjugates. A slight therapeutic effect occurred with either MoHG or ADR alone but treatment with the bispecific antibody followed by the administration of ADR or with the MoHG.Alb.ADR conjugates significantly prolonged survival, with 60% of the treated animals being "tumor free" when sacrificed on day 80. Lower serum concentrations of alphafetoprotein were observed with the bispecific antibody and drug treatment. This suggests that the bispecific antibody/drug treatment is potentially more beneficial in the suppression of distant metastases than the MoHG.Alb.ADR conjugate. This may be due to an increase in the local drug concentration of unmodified adriamycin. 相似文献
19.
H S Cheong J S Chang J M Park S M Byun 《Biochemical and biophysical research communications》1990,173(3):795-800
For the enhancement of antibody binding affinity, a bispecific antibody against two different epitopes in human chorionic gonadotropin hormone, one is in alpha-subunit and the other is in beta-subunit, was prepared by chemical recombination using 5,5'-dithiobis(2-nitrobenzoic acid). The epitopes recognized by antibodies were investigated by competitive radioimmunoassay, two-site sandwich radioimmunoassay and additivity assay and a proper epitope pair was chosen for preparation of the bispecific antibody. This bispecific antibody has dual specificity and as much as 17.2-fold higher affinity than that of monoclonal antibody with higher affinity by dual antigen binding radioimmunoassay and Scatchard plot analysis. 相似文献
20.
Juergen M. Schanzer Katharina Wartha Rebecca Croasdale Samuel Moser Klaus-Peter Künkele Carola Ries Werner Scheuer Harald Duerr Sandra Pompiati Jan Pollman Jan Stracke Wilma Lau Stefan Ries Ulrich Brinkmann Christian Klein Pablo Umana 《The Journal of biological chemistry》2014,289(27):18693-18706
In the present study, we have developed a novel one-arm single chain Fab heterodimeric bispecific IgG (OAscFab-IgG) antibody format targeting the insulin-like growth factor receptor type I (IGF-1R) and the epidermal growth factor receptor (EGFR) with one binding site for each target antigen. The bispecific antibody XGFR is based on the “knob-into-hole” technology for heavy chain heterodimerization with one heavy chain consisting of a single chain Fab to prevent wrong pairing of light chains. XGFR was produced with high expression yields and showed simultaneous binding to IGF-1R and EGFR with high affinity. Due to monovalent binding of XGFR to IGF-1R, IGF-1R internalization was strongly reduced compared with the bivalent parental antibody, leading to enhanced Fc-mediated cellular cytotoxicity. To further increase immune effector functions triggered by XGFR, the Fc portion of the bispecific antibody was glycoengineered, which resulted in strong antibody-dependent cell-mediated cytotoxicity activity. XGFR-mediated inhibition of IGF-1R and EGFR phosphorylation as well as A549 tumor cell proliferation was highly effective and was comparable with a combined treatment with EGFR (GA201) and IGF-1R (R1507) antibodies. XGFR also demonstrated potent anti-tumor efficacy in multiple mouse xenograft tumor models with a complete growth inhibition of AsPC1 human pancreatic tumors and improved survival of SCID beige mice carrying A549 human lung tumors compared with treatment with antibodies targeting either IGF-1R or EGFR. In summary, we have applied rational antibody engineering technology to develop a heterodimeric OAscFab-IgG bispecific antibody, which combines potent signaling inhibition with antibody-dependent cell-mediated cytotoxicity induction and results in superior molecular properties over two established tetravalent bispecific formats. 相似文献