Efficient affinity maturation of antibodies in an engineered chicken B cell line DT40-SW by increasing point mutation(CELL AND TISSUE ENGINEERING)
スポンサーリンク
概要
- 論文の詳細を見る
The chicken B cell line DT40 undergoes hypermutation of immunoglobulin variable region (IgV) genes during culture, thereby constituting an antibody (Ab) library. We previously established an in vitro Ab generation system using an engineered line DT40-SW whose hypermutation machinery can be switched on and off. Abs for various antigens (Ags) can be obtained from the DT40-SW library and the specificity of the Ag-specific clones can be stabilized by stopping hypermutation. Furthermore, the affinity of obtained monoclonal Abs (mAbs) can be improved through further mutation followed by selection, a process analogous to "affinity maturation" that occurs in vivo. Although gene conversion dominantly diversifies the IgV genes in DT40 cells, point mutation is considered to be more favorable for fine-tuning Ab properties during affinity maturation. Here, we examined whether affinity maturation occurs more efficiently when the hypermutation pattern was transformed from gene conversion into point mutation in DT40-SW cells. To this end, we disrupted the XRCC3 gene that is essential for gene conversion. It was found that hemizygous disruption of the XRCC3 gene was sufficient to increase the point mutation frequency. Since hemizygous disruption is conducted more easily, we tested whether the XRCC3 (+/-) mutant generates high-affinity Abs through affinity maturation more efficiently than the wild type. Using this affinity maturation technique, we generated an improved 4-hydroxy-3-nitrophenylacetyl-specific mAb with 〜600-fold lower K_D than that of the original mAb. Taken together, hemizygous disruption of the XRCC3 gene is considered to be useful for obtaining high-affinity mAbs from DT40-SW cells though affinity maturation.
著者
-
Kajita Masamichi
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Magari Masaki
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Todo Kagefumi
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Kanayama Naoki
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Ohmori Hitoshi
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Okazawa Takahiro
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Ikeda Mika
Department of Bioscience and Biotechnology, Graduate School of Natural Science and Technology, Okaya
-
Kanayama Naoki
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
-
Ikeda Mika
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
-
Ikeda Mika
岡山大学 大学院自然科学研究科生物機能工学科
-
Todo Kagefumi
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
-
Magari Masaki
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
-
Ohmori Hitoshi
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
-
Okazawa Takahiro
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
-
Kajita Masamichi
Department Of Bioscience And Biotechnology Graduate School Of Natural Science And Technology Okayama
関連論文
- Conditional transformation of immunoglobulin mutation pattern from gene conversion into point mutation by controlling XRCC3 expression in the DT40 B cell line(CELL AND TISSUE ENGINEERING)
- Efficient affinity maturation of antibodies in an engineered chicken B cell line DT40-SW by increasing point mutation
- The Vanstone-Zuccherato Schemes Revisited(Information Security)
- Efficient affinity maturation of antibodies in an engineered chicken B cell line DT40-SW by increasing point mutation(CELL AND TISSUE ENGINEERING)
- Optimised Versions of the Ate and Twisted Ate Pairings
- Conditional transformation of immunoglobulin mutation pattern from gene conversion into point mutation by controlling XRCC3 expression in the DT40 B cell line
- Enhancement of antibody production from a chicken B cell line DT40 by reducing Pax5 expression(MEDICAL BIOTECHNOLOGY)
- Novel In Vitro Screening System for Monoclonal Antibodies Using Hypermutating Chicken B Cell Library(METHOD)
- ACTIVATION OF MURINE LYMPHOCYTES BY 2-MERCAPTOETHANOL AND RELATED THIOL COMPOUNDS AND ITS MECHANISM. II. EFFECT OF THIOL COMPOUNDS ON THE VIABILITY OF THE LYMPHOCYTES UNDER A CYSTINE-DEFICIENT CULTURE CONDITION
- IN VITRO GENERATION OF ANTIGEN-SPECIFIC SUPPRESSOR T CELL ACTIVITY. CULTURE CONDITIONS FOR ABROGATING THE INDUCTION OF NONSPECIFIC SUPPRESSOR T CELLS
- Optimised Versions of the Ate and Twisted Ate Pairings
- Follow-up results of arthroscopic synovectomy for the rheumatoid knee
- Enhancement of Transgene Expression in Mammalian Cell Line by a Δ^7-Prostaglandin A_1 Analogue
- High mobility group box-1-inducible melanoma inhibitory activity is associated with nodal metastasis and lymphangiogenesis in oral squamous cell carcinoma
- Small Bowel Obstruction Caused by Anisakiasis of the Small Intestine : Report of a Case
- Transmesosigmoid Hernia : Report of a Case
- THE USE OF β-CYCLODEXTRIN IN LYMPHOCYTE CULTURE. INDUCTION OF THE PRIMARY ANTIBODY RESPONSE IN VITRO IN CALF SERUM- OR NEWBORN BOVINE SERUM-CONTAINING CULTURE MEDIUM
- Infection-induced urethral defect treated by urethral reconstruction with a radial forearm flap
- DEGRADATION OF HISTAMINE IN THE PRESENCE OF ASCORBIC ACID AND Cu^ ION : INVOLVEMENT OF HYDROGEN PEROXIDE
- Genomic Structures and Chromosomal Location of p91, a Novel Murine Regulatory Receptor Family^1
- 5-Azacytidine Enhances Transient Expression of a Transfected Gene in Cultured Mammalian Cells
- Enhancement of Expression of the Introduced Gene by 5-Azacytidine in Mammalian Cell Lines
- Methionine-enkephalin secreted by human colorectal cancer cells suppresses T lymphocytes
- Characterization of ASK Mice, a Strain Highly Sensitive to Anaphylactic Shock
- Suppression of in Vitro Cellular Immune Response by Nitrogen-Containing Terpene Alcohol Derivatives
- Enhancement of Expression of Introduced β-Galactosidase Gene by 2-Aminopurine in Mammalian Cell Lines
- Analysis of Baby-Step Giant-Step Algorithms for Non-uniform Distributions (Fundamental) (Cryptography and Information Security)
- Generating Secure Genus Two Hyperelliptic Curves Using Elkies' Point Counting Algorithm
- EFFECT OF N-(2-CARBOXYPHENYL)-4-CHLOROANTHRANILIC ACID DISODIUM SALT (CCA) ON THE INDUCTION OF HELPER AND SUPPRESSOR T CELLS IN VITRO AND IN VIVO
- Production of an Immunosuppressive Polysaccharide, AZ9, in the Culture of Klebsiella oxytoca Strain TNM3
- Search for Immunosuppressive Substances Produced in the Culture of Enteric Bacteria Isolated from Marine Fish
- Subconjunctival Immunization of Mice for Inducing IgE Antibody Response in Parotic Lymph Node
- Scalar Multiplication on Pairing Friendly Elliptic Curves
- Augmentation of the antibody response by lipoic acid in mice. I. Analysis of the mode of action in an in vitro culture system.
- A Note on the Pairing Computation Using Normalized Miller Functions
- Advanced glycation end products (AGE) induce the receptor for AGE in the colonic mucosa of azoxymethane-injected Fischer 344 rats fed with a high-linoleic acid and high-glucose diet
- Phorbol Ester-Induced Reversible Inactivation of Cytotoxic T Cell Function: Correlation with Down-Regulation of Protein Kinase C Activity.
- Implementation of an Elliptic Curve Scalar Multiplication Method Using Division Polynomials
- Demonstration of the Requirements for Cholesterol or Low Density Lipoprotein in the Antibody Response of Murine Lymphocytes in Serum-free Culture
- The use of .BETA.-cyclodextrin-containing culture medium for in vitro evaluation of immunomodulating agents.
- Augmentation of in vitro antibody response by disulfide compounds. I. Comparison between intermolecular and intramolecular disulfides.
- Augmentation of the antibody response by lipoic acid in mice. II Restoration of the antibody response in immunosuppressed mice.