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Construction and selection of human anti-idiotypic antibody single chain variable fragments or CDR3 fragments of nasopharyngeal carcinoma.
J Exp Clin Cancer Res. 2004 Dec; 23(4):607-15.JE

Abstract

Peripheral blood mononuclear cells (PBMCs) of patients with NPC were immunized in vitro by anti-NPC monoclonal antibody FC2 and transformed by Epstein-Barr virus (EBV). Detection showed that of 10 NPC patients, 8 patients' B cells immunized by FC2 and transformed by EBV produced anti-idiotypic antibodies to NPC. Five types of VH genes and 7 types of VL genes were obtained by RT-PCR amplification and then connected with (Gly4Ser)3 linker to form 14 types of scFv genes. ScFv genes digested with Sfi I were cloned into vector fUSE5 and transformed into E. coli MC 1061. Phage anti-idiotypic antibody library with 1.5 x 10(8) clones was obtained. After four rounds of panning, 270 phage clones were selected randomly and 91 FC2-positive clones were obtained by Sandwich ELISA, the positive ratio was 33.7%. 5 clones (D83, E92, G22, I50, I54), which might display beta type Ab2 scFv, were selected by binding inhibition test. These 5 phage anti-idiotypic antibodies were further analyzed by DNA sequencing. The VDJ regions of G22, I50, I54 belonged to VH4-39-D4-11-JH3-linker-V1-19-JL2, VH4-4-D4-11-JH6 and VH4-31-D4-11-JH6, respectively. E92 had the same VDJ regions with G22; D83 had the same VDJ regions with 150. So, a strategy for preparing and selecting beta type Ab2 scFv or CDR by means of immunization in vitro, EBV transformation and phage display technique is feasible, which paves a way for preparing cancer vaccine using beta type Ab2 scFv.

Authors+Show Affiliations

Cancer Research Institute, Xiang-Ya School of Medicine, Central South University, China.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15743031

Citation

He, X, et al. "Construction and Selection of Human Anti-idiotypic Antibody Single Chain Variable Fragments or CDR3 Fragments of Nasopharyngeal Carcinoma." Journal of Experimental & Clinical Cancer Research : CR, vol. 23, no. 4, 2004, pp. 607-15.
He X, Li G, Zhu J. Construction and selection of human anti-idiotypic antibody single chain variable fragments or CDR3 fragments of nasopharyngeal carcinoma. J Exp Clin Cancer Res. 2004;23(4):607-15.
He, X., Li, G., & Zhu, J. (2004). Construction and selection of human anti-idiotypic antibody single chain variable fragments or CDR3 fragments of nasopharyngeal carcinoma. Journal of Experimental & Clinical Cancer Research : CR, 23(4), 607-15.
He X, Li G, Zhu J. Construction and Selection of Human Anti-idiotypic Antibody Single Chain Variable Fragments or CDR3 Fragments of Nasopharyngeal Carcinoma. J Exp Clin Cancer Res. 2004;23(4):607-15. PubMed PMID: 15743031.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction and selection of human anti-idiotypic antibody single chain variable fragments or CDR3 fragments of nasopharyngeal carcinoma. AU - He,X, AU - Li,G, AU - Zhu,J, PY - 2005/3/4/pubmed PY - 2005/6/23/medline PY - 2005/3/4/entrez SP - 607 EP - 15 JF - Journal of experimental & clinical cancer research : CR JO - J Exp Clin Cancer Res VL - 23 IS - 4 N2 - Peripheral blood mononuclear cells (PBMCs) of patients with NPC were immunized in vitro by anti-NPC monoclonal antibody FC2 and transformed by Epstein-Barr virus (EBV). Detection showed that of 10 NPC patients, 8 patients' B cells immunized by FC2 and transformed by EBV produced anti-idiotypic antibodies to NPC. Five types of VH genes and 7 types of VL genes were obtained by RT-PCR amplification and then connected with (Gly4Ser)3 linker to form 14 types of scFv genes. ScFv genes digested with Sfi I were cloned into vector fUSE5 and transformed into E. coli MC 1061. Phage anti-idiotypic antibody library with 1.5 x 10(8) clones was obtained. After four rounds of panning, 270 phage clones were selected randomly and 91 FC2-positive clones were obtained by Sandwich ELISA, the positive ratio was 33.7%. 5 clones (D83, E92, G22, I50, I54), which might display beta type Ab2 scFv, were selected by binding inhibition test. These 5 phage anti-idiotypic antibodies were further analyzed by DNA sequencing. The VDJ regions of G22, I50, I54 belonged to VH4-39-D4-11-JH3-linker-V1-19-JL2, VH4-4-D4-11-JH6 and VH4-31-D4-11-JH6, respectively. E92 had the same VDJ regions with G22; D83 had the same VDJ regions with 150. So, a strategy for preparing and selecting beta type Ab2 scFv or CDR by means of immunization in vitro, EBV transformation and phage display technique is feasible, which paves a way for preparing cancer vaccine using beta type Ab2 scFv. SN - 0392-9078 UR - https://www.unboundmedicine.com/medline/citation/15743031/Construction_and_selection_of_human_anti_idiotypic_antibody_single_chain_variable_fragments_or_CDR3_fragments_of_nasopharyngeal_carcinoma_ L2 - http://www.diseaseinfosearch.org/result/5110 DB - PRIME DP - Unbound Medicine ER -