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Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display

Standard

Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display. / Dong, Ling; Chen, Suzhen; Bartsch, Udo; Schachner, Melitta.

in: BIOCHEM BIOPH RES CO, Jahrgang 301, Nr. 1, 31.01.2003, S. 60-70.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Dong, L, Chen, S, Bartsch, U & Schachner, M 2003, 'Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display', BIOCHEM BIOPH RES CO, Jg. 301, Nr. 1, S. 60-70.

APA

Dong, L., Chen, S., Bartsch, U., & Schachner, M. (2003). Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display. BIOCHEM BIOPH RES CO, 301(1), 60-70.

Vancouver

Dong L, Chen S, Bartsch U, Schachner M. Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display. BIOCHEM BIOPH RES CO. 2003 Jan 31;301(1):60-70.

Bibtex

@article{1e2d9f27714b47c69e21e4f26c53e083,
title = "Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display",
abstract = "The recognition molecule L1 plays important functional roles in the nervous system and in non-neural tissues. Since antibodies to L1 are of prime importance to study its functional properties, we have generated affinity matured human single chain variable fragment (scFv) antibodies against mouse L1 by introducing random mutations in the complementarity determining regions (CDRs) of a previously isolated scFv antibody heavy chain (CDR1 and CDR2) and light chain (CDR3). After biopanning the mutant library, a clone (5F7) that gave the strongest ELISA signal was expressed, purified, and characterized. The dissociation constant of 5F7 (2.86 x 10(-8)M) was decreased 60-fold compared to the wild type clone G6 (1.72 x 10(-6)M). 5F7 detected L1 by Western blot analysis in mouse brain homogenates and recognized L1 in L1 transfected cells and cryosections from mouse retina and optic nerve by immunofluorescence. Bivalent 5F7 scFv antibody (5F7-Cys) was also generated and showed a dissociation constant of 5.22 x 10(-9)M that is 5.5-fold lower than that of monomeric 5F7 antibody. The bivalent affinity matured L1 scFv antibody thus showed stronger binding by a factor of 310 compared to the wild type clone. This antibody should be useful in various biological assays.",
keywords = "Animals, Antibodies, Antibody Affinity, Binding Sites, Antibody, CHO Cells, Cricetinae, Dimerization, Epitope Mapping, Genetic Vectors, Humans, Immunoglobulin Variable Region, Immunohistochemistry, Mice, Mutagenesis, Site-Directed, Neural Cell Adhesion Molecule L1, Optic Nerve, Peptide Library, Recombinant Fusion Proteins, Retina, Journal Article, Research Support, Non-U.S. Gov't",
author = "Ling Dong and Suzhen Chen and Udo Bartsch and Melitta Schachner",
year = "2003",
month = jan,
day = "31",
language = "English",
volume = "301",
pages = "60--70",
journal = "BIOCHEM BIOPH RES CO",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

RIS

TY - JOUR

T1 - Generation of affinity matured scFv antibodies against mouse neural cell adhesion molecule L1 by phage display

AU - Dong, Ling

AU - Chen, Suzhen

AU - Bartsch, Udo

AU - Schachner, Melitta

PY - 2003/1/31

Y1 - 2003/1/31

N2 - The recognition molecule L1 plays important functional roles in the nervous system and in non-neural tissues. Since antibodies to L1 are of prime importance to study its functional properties, we have generated affinity matured human single chain variable fragment (scFv) antibodies against mouse L1 by introducing random mutations in the complementarity determining regions (CDRs) of a previously isolated scFv antibody heavy chain (CDR1 and CDR2) and light chain (CDR3). After biopanning the mutant library, a clone (5F7) that gave the strongest ELISA signal was expressed, purified, and characterized. The dissociation constant of 5F7 (2.86 x 10(-8)M) was decreased 60-fold compared to the wild type clone G6 (1.72 x 10(-6)M). 5F7 detected L1 by Western blot analysis in mouse brain homogenates and recognized L1 in L1 transfected cells and cryosections from mouse retina and optic nerve by immunofluorescence. Bivalent 5F7 scFv antibody (5F7-Cys) was also generated and showed a dissociation constant of 5.22 x 10(-9)M that is 5.5-fold lower than that of monomeric 5F7 antibody. The bivalent affinity matured L1 scFv antibody thus showed stronger binding by a factor of 310 compared to the wild type clone. This antibody should be useful in various biological assays.

AB - The recognition molecule L1 plays important functional roles in the nervous system and in non-neural tissues. Since antibodies to L1 are of prime importance to study its functional properties, we have generated affinity matured human single chain variable fragment (scFv) antibodies against mouse L1 by introducing random mutations in the complementarity determining regions (CDRs) of a previously isolated scFv antibody heavy chain (CDR1 and CDR2) and light chain (CDR3). After biopanning the mutant library, a clone (5F7) that gave the strongest ELISA signal was expressed, purified, and characterized. The dissociation constant of 5F7 (2.86 x 10(-8)M) was decreased 60-fold compared to the wild type clone G6 (1.72 x 10(-6)M). 5F7 detected L1 by Western blot analysis in mouse brain homogenates and recognized L1 in L1 transfected cells and cryosections from mouse retina and optic nerve by immunofluorescence. Bivalent 5F7 scFv antibody (5F7-Cys) was also generated and showed a dissociation constant of 5.22 x 10(-9)M that is 5.5-fold lower than that of monomeric 5F7 antibody. The bivalent affinity matured L1 scFv antibody thus showed stronger binding by a factor of 310 compared to the wild type clone. This antibody should be useful in various biological assays.

KW - Animals

KW - Antibodies

KW - Antibody Affinity

KW - Binding Sites, Antibody

KW - CHO Cells

KW - Cricetinae

KW - Dimerization

KW - Epitope Mapping

KW - Genetic Vectors

KW - Humans

KW - Immunoglobulin Variable Region

KW - Immunohistochemistry

KW - Mice

KW - Mutagenesis, Site-Directed

KW - Neural Cell Adhesion Molecule L1

KW - Optic Nerve

KW - Peptide Library

KW - Recombinant Fusion Proteins

KW - Retina

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

M3 - SCORING: Journal article

C2 - 12535641

VL - 301

SP - 60

EP - 70

JO - BIOCHEM BIOPH RES CO

JF - BIOCHEM BIOPH RES CO

SN - 0006-291X

IS - 1

ER -