Objective To investigate the killing effect of humanized antibodies targeting tissue factors on colon cancer cells as well as migration-inhibiting effect. Methods Humanized anti-tissue factor antibody was purified by Protein A and gel filtration chromatography from cultured CHO-5G4.1 cells that highly express the antibody. The purity was detected by capillary SDS-PAGE. Anticoagulant activity was assessed using the prothrombin time test. Killing effect of the antibody on SW620 and SW480 colorectal cancer cells was tested using antibody-dependent cell-mediated cytotoxicity (ADCC). The effect of antibodies on cell migration was investigated using Transwell assay. Gelatin zymography and Western blotting were used to detect the changes of matrix metalloproteinase 2 (MMP2), MMP9, focal adhesion kinase (FAK) and phosphorylated FAK (p-FAK) after treatment with humanized anti-tissue factor antibody. Results The purity of the humanized anti-tissue factor antibody was estimated 96.9% by the two-step method. The purified antibody showed an obvious anticoagulant activity. The antibody treatment had a significant killing effect on colorectal cancer cells through ADCC and a significant inhibiting effect (99%) on cell migration. The antibody significantly inhibited expression of MMP2 and p-FAK. Conclusion The humanized anti-tissue factor antibody can effectively kill tumor cells through ADCC and inhibit cancer cell migration, which is possibly mediated by MMP2 expression through suppression of FAK signaling.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Concizumab prophylaxis in people with haemophilia A or haemophilia B without inhibitors (explorer8): a prospective, multicentre, open-label, randomised, phase 3a trial.
Lancet Haematol
December 2024
Institute of Clinical Hemostaseology and Transfusion Medicine, Saarland University and University Hospital, Homburg, Germany.
Background: Concizumab is an anti-tissue factor pathway inhibitor monoclonal antibody in development as a once-daily, subcutaneous prophylaxis for patients with haemophilia A or haemophilia B with or without inhibitors. We aimed to assess the efficacy and safety of concizumab in patients with haemophilia A or B without inhibitors. Here we report the results from the confirmatory analysis cutoff.
View Article and Find Full Text PDFAnti-tissue factor antibody conjugated with monomethyl auristatin E or deruxtecan in pancreatic cancer models.
Cancer Sci
December 2024
Division of Developmental Therapeutics, EPOC, National Cancer Center, Kashiwa, Japan.
Antibody-drug conjugates (ADCs) have been recognized as a promising class of cancer therapeutics. Tissue factor (TF), an initiator of the blood coagulation pathway, has been investigated regarding its relationship with cancer, and several preclinical and clinical studies have presented data on anti-TF ADCs, including tisotumab vedotin, which was approved in 2021. However, the feasibility of other payloads in the design of anti-TF ADCs is still unclear because no reports have compared payloads with different cytotoxic mechanisms.
View Article and Find Full Text PDFMinimal interference of concizumab with standard clinical coagulation laboratory assays - An in vitro study.
Haemophilia
July 2024
Rare Disease Research, Global Drug Discovery, Novo Nordisk A/S, Måløv, Denmark.
Introduction: Non-factor replacement therapies are emerging as prophylactic treatment options in haemophilia A or B (HA/HB) with and without inhibitors. Concizumab is an anti-tissue factor pathway inhibitor (TFPI) monoclonal antibody preventing factor (F)Xa inhibition and enhancing thrombin generation. Based on experience with other non-factor therapies and extended half-life products, there is a focus on potential interference with common clinical coagulation assays used to monitor patients treated with concizumab.
View Article and Find Full Text PDFPhase 3 Trial of Concizumab in Hemophilia with Inhibitors.
N Engl J Med
August 2023
From the Department of Transfusion Medicine, Nagoya University Hospital, Nagoya (T.M.), and Nara Medical University, Kashiwara (K.N.) - both in Japan; Indiana Hemophilia and Thrombosis Center, Indianapolis (A.S.); the Department of Hematology, Christian Medical College, Vellore, India (A.A.); the Center for Bleeding Disorders and Coagulation, Department of Oncology, Careggi University Hospital, Florence, Italy (P.A.); the Division of Hematology, Department of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand (G.C.); Novo Nordisk, Søborg, Denmark (K.C., J.H., S.H.J., J.S.R.); the Reference Center for Hemophilia and Rare Congenital Bleeding Disorders, Bicêtre Hospital Assistance Publique-Hôpitaux de Paris, University of Paris-Saclay and UMR_S1176 INSERM, Le Kremlin-Bicêtre, France (R.O.); the Department of Pediatrics, University of Texas Health Long School of Medicine, San Antonio (M.F.-J.); the Department of Medicine, Hospital Pulau Pinang, Georgetown, Malaysia (A.-S.G.); the Department of Molecular Medicine and Haematology, University of the Witwatersrand, National Health Laboratory Service, Johannesburg (J.M.); Great Ormond Street Hospital for Children NHS Foundation Trust, London (M.M.); the Institute of Blood Pathology and Transfusion Medicine, Lviv (O.S.), and National Specialized Children's Hospital Okhmatdyt, Kyiv (K.V.) - both in Ukraine; the Ronald Sawers Haemophilia Centre,Alfred Hospital, and the Australian Centre for Blood Diseases, Monash University - both in Melbourne, VIC, Australia (H.T.); Dr. José Eleuterio González Monterrey University Hospital, Monterrey, México (L.V.M.); the Department of Hemostasis Disorders and Internal Medicine, Laboratory of Hemostasis and Metabolic Diseases, Institute of Hematology and Transfusion Medicine, Warsaw, Poland (J.W.); the Department of Pediatrics, Daejeon Eulji Medical Center, Eulji University School of Medicine, Daejeon, South Korea (C.W.Y.); the National Research Center for Hematology of the Ministry of Health of the Russian Federation, Moscow (N.Z.); the Division of Pediatric Hematology-Oncology, Istanbul University Oncology Institute, Istanbul, Turkey (B.Z.); and the Hematology Department, La Paz University Hospital, Hospital La Paz Institute for Health Research, Universidad Autónoma Madrid, Madrid (V.J.-Y.).
Background: Concizumab is an anti-tissue factor pathway inhibitor monoclonal antibody designed to achieve hemostasis in all hemophilia types, with subcutaneous administration. A previous trial of concizumab (explorer4) established proof of concept in patients with hemophilia A or B with inhibitors.
Methods: We conducted the explorer7 trial to assess the safety and efficacy of concizumab in patients with hemophilia A or B with inhibitors.
Evolution of Antidrug Antibody Assays During the Development of Anti-Tissue Factor Pathway Inhibitor Monoclonal Antibody Marstacimab.
AAPS J
August 2023
Rare Disease Research Unit, Pfizer Worldwide Research & Development, 610 Main St., 2nd Floor, Cambridge, Massachusetts, 02139, USA.
Tissue factor pathway inhibitor (TFPI) is an endogenous inhibitor of the extrinsic coagulation pathway. In patients with hemophilia A or B, inhibition of TFPI is an alternative therapeutic approach that augments the extrinsic coagulation pathway. Marstacimab is an investigational fully human monoclonal antibody that binds and neutralizes TFPI and is being evaluated as a prophylactic treatment to prevent or reduce the frequency of bleeding episodes in patients with severe hemophilia A or B, with or without inhibitors (antibodies against coagulation factors).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!