Comparison of Humanized IgG and FvFc Anti-CD3 Monoclonal Antibodies Expressed in CHO Cells
Mol. biotechnol
; 45(3): 218-225, Mar 25,2010.
Article
in English
| Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP, SESSP-IBACERVO
| ID: biblio-1064783
Responsible library:
BR78.1
Localization: BR78.1
ABSTRACT
Two humanized monoclonal antibody constructs bearing the same variable regions of an anti-CD3 monoclonal antibody, whole IgG and FvFc, were expressed in CHO cells. Random and site-specific integration were used resulting in similar expression levels. The transfectants were selected with appropriate selection agent, andthe surviving cells were plated in semi-solid medium for capture with FITC-conjugated anti-human IG antibody andpicked with the robotic ClonePix FL. Conditioned media from selected clones were purified by affinity chromatographyand characterized by SDS-PAGE, Western-blot, SEC-HPLC, and isoelectric focusing. Binding to the targetpresent in healthy human mononuclear cells was assessed by flow cytometry, as well as by competition between thetwo constructs and the original murine monoclonal antibody. The humanized constructs were not able to dislodgethe murine antibody while the murine anti-CD3 antibody could dislodge around 20% of the FvFc or IgG humanizedversions. Further in vitro and in vivo pre-clinical analyses.
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Collection:
National databases
/
Brazil
Database:
Sec. Est. Saúde SP
/
SESSP-IBACERVO
/
SESSP-IBPROD
Main subject:
Cricetulus
/
Cricetinae
/
CHO Cells
/
Antibodies, Monoclonal
Type of study:
Evaluation study
Limits:
Animals
/
Humans
Language:
English
Journal:
Mol. biotechnol
Year:
2010
Document type:
Article
Institution/Affiliation country:
Instituto Butantan/BR