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[Production of phage-displayed anti-idiotypic antibody single chain variable fragments to MG7 monoclonal antibody directed against gastric carcinoma].
Zhonghua Yi Xue Za Zhi. 2001 Jan 10; 81(1):33-6.ZY

Abstract

OBJECTIVE

To generate phage-displayed anti-idiotypic antibody single chain variable fragments (anti-Id ScFv) MG7 to monoclonal antibody directed against gastric carcinoma so as to lay a foundation for developing anti-Id ScFv vaccine of the cancer.

METHODS

Balb/c mice were immunized i.p. with purified MG7 monoclonal antibody conjugated with keyhole limpet hemocyanin. mRNA was isolated from the spleens of immunized mice. Heavy and light chain genes (VH and VL) of antibody were amplified separately and assembled into ScFv genes with a specially constructed linker DNA by RT-PCR. The ScFv genes were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage, which displayed ScFv fragments as a fusion with gene 3 protein on the tips of M13 phage. After four rounds of panning with monoclonal antibody MG7, the MG7-positive clones were selected with the enzyme-linked immunosorbent assay (ELISA) from the enriched phages. The types of the anti-Id ScFv displayed on the selected phage clones were primarily identified by competition ELISA.

RESULTS

The VH, VL and ScFv DNAs were about 340, 320 and 750 bp respectively. Twenty-four MG7-positive clones were selected from 40 enriched phage clones. Five of the 24 clones displayed beta or gamma type anti-Id ScFv.

CONCLUSION

The anti-Id ScFv frangments to MG7 monoclonal antibody can be successfully selected by recombinant phage antibody technique, which paves a way for the study of prevention and cure of gastric carcinoma using anti-Id ScFv.

Authors+Show Affiliations

Institute of Digestive Disease, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.No affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article

Language

chi

PubMed ID

11798849

Citation

He, F, et al. "[Production of Phage-displayed Anti-idiotypic Antibody Single Chain Variable Fragments to MG7 Monoclonal Antibody Directed Against Gastric Carcinoma]." Zhonghua Yi Xue Za Zhi, vol. 81, no. 1, 2001, pp. 33-6.
He F, Nie Y, Han Z. [Production of phage-displayed anti-idiotypic antibody single chain variable fragments to MG7 monoclonal antibody directed against gastric carcinoma]. Zhonghua Yi Xue Za Zhi. 2001;81(1):33-6.
He, F., Nie, Y., & Han, Z. (2001). [Production of phage-displayed anti-idiotypic antibody single chain variable fragments to MG7 monoclonal antibody directed against gastric carcinoma]. Zhonghua Yi Xue Za Zhi, 81(1), 33-6.
He F, Nie Y, Han Z. [Production of Phage-displayed Anti-idiotypic Antibody Single Chain Variable Fragments to MG7 Monoclonal Antibody Directed Against Gastric Carcinoma]. Zhonghua Yi Xue Za Zhi. 2001 Jan 10;81(1):33-6. PubMed PMID: 11798849.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Production of phage-displayed anti-idiotypic antibody single chain variable fragments to MG7 monoclonal antibody directed against gastric carcinoma]. AU - He,F, AU - Nie,Y, AU - Han,Z, PY - 2002/1/19/pubmed PY - 2002/4/16/medline PY - 2002/1/19/entrez SP - 33 EP - 6 JF - Zhonghua yi xue za zhi JO - Zhonghua Yi Xue Za Zhi VL - 81 IS - 1 N2 - OBJECTIVE: To generate phage-displayed anti-idiotypic antibody single chain variable fragments (anti-Id ScFv) MG7 to monoclonal antibody directed against gastric carcinoma so as to lay a foundation for developing anti-Id ScFv vaccine of the cancer. METHODS: Balb/c mice were immunized i.p. with purified MG7 monoclonal antibody conjugated with keyhole limpet hemocyanin. mRNA was isolated from the spleens of immunized mice. Heavy and light chain genes (VH and VL) of antibody were amplified separately and assembled into ScFv genes with a specially constructed linker DNA by RT-PCR. The ScFv genes were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage, which displayed ScFv fragments as a fusion with gene 3 protein on the tips of M13 phage. After four rounds of panning with monoclonal antibody MG7, the MG7-positive clones were selected with the enzyme-linked immunosorbent assay (ELISA) from the enriched phages. The types of the anti-Id ScFv displayed on the selected phage clones were primarily identified by competition ELISA. RESULTS: The VH, VL and ScFv DNAs were about 340, 320 and 750 bp respectively. Twenty-four MG7-positive clones were selected from 40 enriched phage clones. Five of the 24 clones displayed beta or gamma type anti-Id ScFv. CONCLUSION: The anti-Id ScFv frangments to MG7 monoclonal antibody can be successfully selected by recombinant phage antibody technique, which paves a way for the study of prevention and cure of gastric carcinoma using anti-Id ScFv. SN - 0376-2491 UR - https://www.unboundmedicine.com/medline/citation/11798849/[Production_of_phage_displayed_anti_idiotypic_antibody_single_chain_variable_fragments_to_MG7_monoclonal_antibody_directed_against_gastric_carcinoma]_ L2 - http://journal.yiigle.com/LinkIn.do?linkin_type=pubmed&issn=0376-2491&year=2001&vol=81&issue=1&fpage=33 DB - PRIME DP - Unbound Medicine ER -