Abstract
We have previously described a vector system for generating recombinant polyclonal antibody libraries. This system uses bidirectional phagemid and mammalian expression vectors to facilitate mass transfer of selected variable light and variable heavy (VL-VH) region gene pairs from the phagemid to the mammalian vector, to express polyclonal libraries of whole IgG antibodies. We report here the first stage of generating a polyclonal antibody library to the human breast carcinoma cell line BT-20, using this vector system. VL and VH region gene pairs were obtained from a mouse immunized with BT-20 cells, and cloned, in opposite transcriptional orientations, in the bidirectional phagemid vector, to produce an Fab phage display library. This library was selected by panning on BT-20 cells and shown to bind specifically to BT-20 cells. Such libraries, after suitable negative selection to eliminate major reactivities against normal tissue, could be transferred in mass to our bidirectional mammalian expression vector for production of libraries of chimeric antibodies with mouse V regions and human constant (C) regions. These polyclonal antibody libraries will mediate effector functions and are expected to be useful for breast cancer therapy, as well as diagnosis.
Keywords: Polyclonal fab phage display library, Human breast carcinoma cell line BT 20, Immunization of Mice
Combinatorial Chemistry & High Throughput Screening
Title: Generation of a Polyclonal Fab Phage Display Library to the Human Breast Carcinoma Cell Line BT-20
Volume: 3 Issue: 1
Author(s): K. E. Santora, S. Sarantopoulos, W. Den, S. Petersen-Mahrt, S. R. Sompuram and J. Sharon
Affiliation:
Keywords: Polyclonal fab phage display library, Human breast carcinoma cell line BT 20, Immunization of Mice
Abstract: We have previously described a vector system for generating recombinant polyclonal antibody libraries. This system uses bidirectional phagemid and mammalian expression vectors to facilitate mass transfer of selected variable light and variable heavy (VL-VH) region gene pairs from the phagemid to the mammalian vector, to express polyclonal libraries of whole IgG antibodies. We report here the first stage of generating a polyclonal antibody library to the human breast carcinoma cell line BT-20, using this vector system. VL and VH region gene pairs were obtained from a mouse immunized with BT-20 cells, and cloned, in opposite transcriptional orientations, in the bidirectional phagemid vector, to produce an Fab phage display library. This library was selected by panning on BT-20 cells and shown to bind specifically to BT-20 cells. Such libraries, after suitable negative selection to eliminate major reactivities against normal tissue, could be transferred in mass to our bidirectional mammalian expression vector for production of libraries of chimeric antibodies with mouse V regions and human constant (C) regions. These polyclonal antibody libraries will mediate effector functions and are expected to be useful for breast cancer therapy, as well as diagnosis.
Export Options
About this article
Cite this article as:
Santora E. K., Sarantopoulos S., Den W., Petersen-Mahrt S., Sompuram R. S. and Sharon J., Generation of a Polyclonal Fab Phage Display Library to the Human Breast Carcinoma Cell Line BT-20, Combinatorial Chemistry & High Throughput Screening 2000; 3 (1) . https://dx.doi.org/10.2174/1386207003327765
DOI https://dx.doi.org/10.2174/1386207003327765 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Endogenous and Exogenous Ligands of Aryl Hydrocarbon Receptor: Current State of Art
Current Drug Metabolism Histopathological Determinants of Tumor Resistance: A Special Look to the Immunohistochemical Expression of Carbonic Anhydrase IX in Human Cancers
Current Medicinal Chemistry The Role of Neurogenesis in Neurodegenerative Diseases and its Implications for Therapeutic Development
CNS & Neurological Disorders - Drug Targets Nanoemulsions: A Better Approach for Antidiabetic Drug Delivery
Current Diabetes Reviews Etiology of Neuroinflammatory Pathologies in Neurodegenerative Diseases: A Treatise
Current Psychopharmacology Research Advances in Neuroblastoma Immunotherapy
Current Pediatric Reviews From Multiple PAR1 Receptor/Protein Interactions to their Multiple Therapeutic Implications
Current Topics in Medicinal Chemistry Review of Dose-intense Platinum and/or Paclitaxel Containing Chemotherapy in Advanced and Recurrent Epithelial Ovarian Cancer
Current Pharmaceutical Design Combining Angiogenesis Inhibitors with Radiation: Advances and Challenges in Cancer Treatment
Current Pharmaceutical Design 2-Deoxy-D-Ribose, a Downstream Mediator of Thymidine Phosphorylase, Regulates Tumor Angiogenesis and Progression
Anti-Cancer Agents in Medicinal Chemistry Advances in Computational Structure-Based Drug Design and Application in Drug Discovery
Current Topics in Medicinal Chemistry Aptamers: Potential Applications to Pancreatic Cancer Therapy
Anti-Cancer Agents in Medicinal Chemistry Withdrawn: Ag/PdTi3O7 Nano Catalyst Promoted for Developing Novel Indole- Naphthyridine Derivatives as Potent Anticancer Agents
Current Organocatalysis Heat Shock Protein 90 Inhibitors in Oncology
Current Proteomics The Ultrasonographic Diagnosis of Polycystic Ovary Syndrome
Current Medical Imaging Advancement in Obesity Management: Leptin and Adiponectin Patents
Recent Patents on Endocrine, Metabolic & Immune Drug Discovery Solid Lipid Nanoparticles (SLNs) as a Rising Tool in Drug Delivery Science: One Step Up in Nanotechnology
Current Nanoscience Sympathetic Signaling in Angiogenesis: Implications for Cancer Progression
Current Cancer Therapy Reviews Synthesis, Characterization, and Biological Evaluation of Novel Naringenin Derivatives as Anticancer Agents
Current Bioactive Compounds Protection and Restitution of Gut Barrier by Probiotics: Nutritional and Clinical Implications
Current Nutrition & Food Science