Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein

BMC Cancer
Emiliano PavoniOlga Minenkova

Abstract

CEA is a tumor-associated antigen abundantly expressed on several cancer types, including those naturally refractory to chemotherapy. The selection and characterization of human anti-CEA single-chain antibody fragments (scFv) is a first step toward the construction of new anticancer monoclonal antibodies designed for optimal blood clearance and tumor penetration. The human MA39 scFv, selected for its ability to recognize a CEA epitope expressed on human colon carcinomas, was first isolated from a large semi-synthetic ETH-2 antibody phage library, panned on human purified CEA protein. Subsequently, by in vitro mutagenesis of a gene encoding for the scFv MA39, a new library was established, and new scFv antibodies with improved affinity towards the CEA cognate epitope were selected and characterized. The scFv MA39 antibody was affinity-maturated by in vitro mutagenesis and the new scFv clone, E8, was isolated, typed for CEA family member recognition and its CEACAM1, 3 and 5 shared epitope characterized for expression in a large panel of human normal and tumor tissues and cells. The binding affinity of the scFv E8 is in a range for efficient, in vivo, antigen capture in tumor cells expressing a shared epitope of the CEACAM1, 3 and...Continue Reading

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Citations

May 23, 2009·American Journal of Clinical Pathology·Thilo GambichlerAlexander Kreuter
Oct 18, 2015·Applied and Environmental Microbiology·Chi-Hsin LeeYi-Yuan Yang
Dec 13, 2007·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Apr 18, 2019·International Journal of Molecular Sciences·Chia Chiu LimTheam Soon Lim
May 28, 2021·Scientific Reports·Kaho KajiwaraMitsuyoshi Ueda

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Datasets Mentioned

BETA
U937

Methods Mentioned

BETA
flow cytometry
PCR
ELISA
chip
FCS
electrophoresis
confocal microscopy
xenograft
pull down
gel filtration

Software Mentioned

BIAevaluation
Biacore
Cell Quest
Photoshop

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